AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)...AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.展开更多
Human neural stem cell-derived extracellular vesicles exhibit analogous functions to their parental cells,and can thus be used as substitutes for stem cells in stem cell therapy,thereby mitigating the risks of stem ce...Human neural stem cell-derived extracellular vesicles exhibit analogous functions to their parental cells,and can thus be used as substitutes for stem cells in stem cell therapy,thereby mitigating the risks of stem cell therapy and advancing the frontiers of stem cell-derived treatments.This lays a foundation for the development of potentially potent new treatment modalities for ischemic stroke.However,the precise mechanisms underlying the efficacy and safety of human neural stem cell-derived extracellular vesicles remain unclear,presenting challenges for clinical translation.To promote the translation of therapy based on human neural stem cell-derived extracellular vesicles from the bench to the bedside,we conducted a comprehensive preclinical study to evaluate the efficacy and safety of human neural stem cell-derived extracellular vesicles in the treatment of ischemic stroke.We found that administration of human neural stem cell-derived extracellular vesicles to an ischemic stroke rat model reduced the volume of cerebral infarction and promoted functional recovery by alleviating neuronal apoptosis.The human neural stem cell-derived extracellular vesicles reduced neuronal apoptosis by enhancing phosphorylation of phosphoinositide 3-kinase,mammalian target of rapamycin,and protein kinase B,and these effects were reversed by treatment with a phosphoinositide 3-kinase inhibitor.These findings suggest that human neural stem cell-derived extracellular vesicles play a neuroprotective role in ischemic stroke through activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway.Finally,we showed that human neural stem cell-derived extracellular vesicles have a good in vivo safety profile.Therefore,human neural stem cell-derived extracellular vesicles are a promising potential agent for the treatment of ischemic stroke.展开更多
The published article titled“Knockdown of Rap1b Enhances Apoptosis and Autophagy in Gastric Cancer Cells via the PI3K/Akt/mTOR Pathway”has been retracted from Oncology Research,Vol.24,No.5,2016,pp.287–293.DOI:10.37...The published article titled“Knockdown of Rap1b Enhances Apoptosis and Autophagy in Gastric Cancer Cells via the PI3K/Akt/mTOR Pathway”has been retracted from Oncology Research,Vol.24,No.5,2016,pp.287–293.DOI:10.3727/096504016X14648701447779 URL:https://www.techscience.com/or/v24n5/56977 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases.展开更多
Hepatocellular carcinoma(HCC)is the most common primary liver tumor and the third leading cause of cancer-related mortality globally.The phosphatidylinositol-3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of ra...Hepatocellular carcinoma(HCC)is the most common primary liver tumor and the third leading cause of cancer-related mortality globally.The phosphatidylinositol-3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway is critically involved in HCC pathogenesis,stimulating uncontrolled cell proliferation,survival,and tumor progression.The overactivation of this pathway is strongly linked to poor prognosis,making it a crucial target for therapeutic intervention.The oncogenic roles of PI3K/AKT/mTOR components in HCC have been highlighted,noting that class I PI3K deregulation,phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA)upregulation,and mTOR overexpression could be associated with poor HCC outcomes.To the best of our knowledge,this is the first time that the clinical trials investigating PI3K/AKT/mTOR inhibitors in HCC are analyzed.Accordingly,there is a predominance of mTOR inhibitors,with everolimus being the most frequently utilized drug.However,only 10%of studies advanced to phase III or IV,predominantly involving mTOR inhibitors.Challenges such as adverse events like hyperglycemia and bone marrow suppression,as well as the emergence of treatment resistance,have hindered the success of these therapies.Combination therapies,particularly those involving mitogen-activated protein kinase kinase(MEK)inhibitors,chemotherapy,immune checkpoint inhibitors,and vascular endothelial growth factor(VEGF)inhibitors,have shown promise in overcoming these challenges.Recent advances in nanotechnology offer the potential for improving drug delivery and reducing toxicity.展开更多
Background:Oral squamous cell carcinoma(OSCC)is a common malignant tumor.Recently,Laminin Gamma 2(LAMC2)has been shown to be abnormally expressed in OSCC;however,how LAMC2 signaling contributes to the occurrence and d...Background:Oral squamous cell carcinoma(OSCC)is a common malignant tumor.Recently,Laminin Gamma 2(LAMC2)has been shown to be abnormally expressed in OSCC;however,how LAMC2 signaling contributes to the occurrence and development of OSCC and the role of autophagy in OSCC has not been fully explored.This study aimed to analyze the role and mechanism of LAMC2 signaling in OSCC and the involvement of autophagy in OSCC.Methods:To explore the mechanism by which LAMC2 is highly expressed in OSCC,we used small interfering RNA(siRNA)to knock down LAMC2 to further observe the changes in the signaling pathway.Furthermore,we used cell proliferation assays,Transwell invasion assays,and wound-healing assays to observe the changes in OSCC proliferation,invasion,and metastasis.RFP-LC3 was used to detect the level of autophagy intensity.A cell line-derived xenograft(CDX)model was used to detect the effect of LAMC2 on tumor growth in vivo.Results:This study found that the level of autophagy was correlated with the biological behavior of OSCC.The downregulation of LAMC2 activated autophagy and inhibited OSCC proliferation,invasion,and metastasis via inhibiting the PI3K/AKT/mTOR pathway.Moreover,autophagy has a dual effect on OSCC,and the synergistic downregulation of LAMC2 and autophagy can inhibit OSCC metastasis,invasion,and proliferation via the PI3K/AKT/mTOR pathway.Conclusions:LAMC2 interacts with autophagy to regulate OSCC metastasis,invasion,and proliferation via the PI3K/AKT/mTOR pathway.LAMC2 down-regulation can synergistically modulate autophagy to inhibit OSCC migration,invasion,and proliferation.展开更多
We evaluated the effect of isoquercetin(quercetin-O-3-glucoside-quercetin,IQ)as a functional component of Abeliophyllum disistichum Nakai ethanol extract(ADLE)on prostate cell proliferation and apoptosis and its effec...We evaluated the effect of isoquercetin(quercetin-O-3-glucoside-quercetin,IQ)as a functional component of Abeliophyllum disistichum Nakai ethanol extract(ADLE)on prostate cell proliferation and apoptosis and its effects on the IGF-1/PI3K/Akt/mTOR pathway in benign prostatic hyperplasia(BPH).Metabolites in ADLE were analyzed using UHPLC-qTOF-MS and HPLC.IQ was orally administered(1 or 10 mg/kg)to a testosterone propionate-induced BPH rat model,and its effects on the prostate weight were evaluated.The effect of IQ on androgen receptor(AR)signaling was analyzed in LNCaP cells.Whether IGF-1 and IQ affect the IGF-1/PI3K/Akt/mTOR pathway in BPH-1 cells was also examined.The metabolites in ADLE were identified and quantified,which confirmed that ADLE contained abundant IQ(20.88 mg/g).IQ significantly reduced the prostate size in a concentration-dependent manner in a BPH rat model,and significantly decreased the expression of AR signaling factors in the rat prostate tissue and LNCaP cells in a concentration-dependent manner.IQ also inhibited the PI3K/AKT/mTOR pathway activated by IGF-1 treatment in BPH-1 cells.In BPH-1 cells,IQ led to G0/G1 arrest and suppressed the expression of proliferation factors while inducing apoptosis.Thus,IQ shows potential for use as a pharmaceutical and nutraceutical for BPH.展开更多
A kind of triterpene glycosides echinoside A(EA)was extracted from sea cucumber Pearsonothuria graeffei,and its yield was about 0.78%.The purity of EA was 99.0%,and its molecular weight was 1206 Da.EA was a linear tet...A kind of triterpene glycosides echinoside A(EA)was extracted from sea cucumber Pearsonothuria graeffei,and its yield was about 0.78%.The purity of EA was 99.0%,and its molecular weight was 1206 Da.EA was a linear tetrasaccharide attached to a pentacyclic triterpene aglycon.It inhibited the growth of MDA-MB-231 cells in vitro.The antitumor effect was related to elevate ROS level,decrease mitochondrial membrane potential,enhance caspase-3 expression,induce cells apoptosis and arrest cell cycle at G2/M phase.EA also dose-dependently suppressed the expressions of phophorylation proteins p-PI3K,p-Akt,and p-mTOR as analyzed by western blotting.These results suggested that EA caused MDA-MB-231 cells apoptosis via intrinsic mitochondrial and PI3K/Akt/mTOR pathway.EA can be a potential anti-breast cancer agent to enhance the clinical efficacy.展开更多
Objective To investigate the effects of Niuhuang(Bovis Calculus,BC)and Shexiang(Moschus)(BC-Moschus)on human hepatocellular carcinoma(HCC)cells SMMC-7721 and a nude mouse model of subcutaneous xenografts,and to explor...Objective To investigate the effects of Niuhuang(Bovis Calculus,BC)and Shexiang(Moschus)(BC-Moschus)on human hepatocellular carcinoma(HCC)cells SMMC-7721 and a nude mouse model of subcutaneous xenografts,and to explore its anti-HCC mechanism.Methods The BC-Moschus combination was applied to two liver cancer models in vivo and in vitro.SMMC-7721 was divided into the BC-Moschus group and the control group,and different doses(rude drug dosage 0.625,1.25,2.5,and 5 mg/m L)of BC-Moschus extract were used for the intervention.The proliferation ability of HCC cells was detected using the Cell Counting Kit-8(CCK-8)assay,and the migration ability was detected by a wound healing assay.A subcutaneous xenograft model was prepared using nude mice with human HCC.Specific pathogen-free-grade BALB/c nude mice(5-week-old)were randomly divided into the following groups(n=6 per group):control(0.9%physiological saline 0.2 m L/d),BC-Moschus[BC 45.5 mg/(kg·d)+Moschus 13 mg/(kg·d)],and cisplatin(DDP,intraperitoneal injection5 mg/kg per week)groups.All groups were administered for 14 d.The volume and mass of the subcutaneous xenografts in nude mice were observed.The expression levels of phosphatidylinositol-3 kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway,apoptosis-associated factor p70 S6 Kinase(S6K),Bax,Bcl-2,caspase-3,and caspase-9 in nude mice subcutaneous xenografts were measured by real-time quantitative PCR(RT-qPCR)and Western blot.Terminal Deoxynucleotidy Transferase-Mediated d UTP NickEnd Labeling(TUNEL)was used for quantitative analysis of apoptotic cells.Results The CCK-8 assay demonstrated that the BC-Moschus combination inhibited HCC cell proliferation in a superior manner to the use of BC and Moschus alone,and the inhibition effect was dose-and time-dependent(P<0.01).The wound healing assay showed that the BC-Moschus combination inhibited HCC cell migration(P<0.01).In the subcutaneous xenograft model of nude mice with human HCC,we found that the tumor volume and weight of the BC-Moschus group were lower than those of the control group(P<0.01).The levels of the PI3K/AKT/m TOR signaling pathway and S6K protein in the BC-Moschus and DDP groups were significantly decreased(P<0.01).The expression level of the anti-apoptotic gene Bcl-2 was downregulated(P<0.05),and the expression of the pro-apoptotic gene Baxand apoptosis-related factors caspase-3 and caspase-9 were significantly upregulated(P<0.01).The TUNEL assays further confirmed that the combination of the BC-Moschuas could promote HCC(P<0.01).Conclusion The BC-Moschus combination inhibited the proliferation and migration ability of HCC cells SMMC-7721 and effectively inhibited the growth of subcutaneous xenografts in nude mice.The mechanism may be closely related to the downregulation of the PI3K/AKT/mTOR pathway,regulation of apoptosis-related protein caspase-3,caspase-9,Bcl-2,and Bax expression,and promotion of apoptosis.展开更多
Catechins have been proven to exert antitumor effects in different kinds of cancers.However,the underlying mechanisms have not been completely clarified yet.This study aimed to assess the effects and mechanisms of(-)-...Catechins have been proven to exert antitumor effects in different kinds of cancers.However,the underlying mechanisms have not been completely clarified yet.This study aimed to assess the effects and mechanisms of(-)-epigallocatechin-3-gallate(EGCG)and(-)-epicatechin-3-gallate(ECG)on human melanoma skin A375 cells.Results showed that EGCG and ECG inhibited the proliferation of A375 cells and ECG showed better inhibitory effect.Flow cytometry analysis had shown that EGCG and ECG induced apoptosis and led to cell cycle arrest.EGCG and ECG decreased Bcl-2 expression and upregulated Caspase-3 protein level,indicating the development of apoptosis.Furthermore,EGCG and ECG could decreased mitochondrial membrane potential of A375 cells.In addition,the expression of Beclin-1,LC3 and Sirt3 were downregulated at protein levels,which known to be associated with autophagy.After autophagy was increased by rapamycin,the apoptotic trend was not change,indicating that apoptosis and autophagy are independent.Mechanistically,EGCG and ECG treatments decreased phosphorylated-AMPK(p-AMPK)and increased the ratios of P-PI3K,p-AKT and p-mTOR in melanoma cells.Conclusively,EGCG and ECG induced apoptosis via mitochondrial signaling pathway,downregulated autophagy through modulating the AMPK/mTOR and PI3K/AKT/mTOR signaling pathway.It indicated that EGCG and ECG may be utilized in human melanoma treatment.展开更多
Monascus vinegar(MV) is a typical fermented food with various health-promoting effects. This study aimed to evaluate the role of MV in alleviating high-fat-diet-induced inflammation in rats with hyperlipidemia and elu...Monascus vinegar(MV) is a typical fermented food with various health-promoting effects. This study aimed to evaluate the role of MV in alleviating high-fat-diet-induced inflammation in rats with hyperlipidemia and elucidate the possible regulatory mechanisms. In the study, serum lipid profiles, liver pathology and liver inflammatory cytokines were analyzed in hyperlipidemia rats with MV(0.5 mL/kg mb, 2 mL/kg mb). Results showed that the administration of MV alleviated dyslipidemia by decreasing the serum and liver levels of triglyceride and total cholesterol. Increase in hepatic lipase and carnitine palmitoyl transferase 1(CPT-1)levels and decrease in hepatocyte steatosis, nephritis, and intestinal tissue injury in the HD group showed that high-dose MV can significantly suppress hepatic lipid accumulation and steatosis. In addition, compared with the model(MOD) group, the HD group showed significantly down-regulated the level of serum or hepatic alanine aminotransferase(ALT), aspartate aminotransferase(AST), CPT-1, interleukin(IL)-2, IL-6, IL-12,and tumor necrosis factor α(TNF-α). Moreover, the HD group showed repressed hepatic nuclear factor κB(NF-κB) pathway and inactivated phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway mitigated liver inflammation. Similar results were obtained from cell experiments. Collectively, these findings revealed that MV might attenuate high-fat-diet-induced inflammation by inhibiting the NF-κB and PI3K/Akt/mTOR pathways.展开更多
As a pathway that plays a role in nutrient absorption,anabolic response,cell growth and survival,the important role of AKT/mTOR in tumorigenesis has also come to light.For cancer patients,most deaths are caused by the...As a pathway that plays a role in nutrient absorption,anabolic response,cell growth and survival,the important role of AKT/mTOR in tumorigenesis has also come to light.For cancer patients,most deaths are caused by the growth of metastatic tumors outside the primary focus.Therefore,migration and invasion in the late stage of tumor progression are the main unresolved issues in the study of tumor pathogenesis,and AKT/mTOR has been found to participate in the migration and invasion of cancer cells,which means that the study of this pathway may contribute to a solution for the problem.Because of its extensive and complex functions in the organism,this pathway can be regulated by a variety of different signals in the body,and then realize its function through different downstream signal molecules.This article reviews the proteins that can indirectly affect this pathway by regulating the common upstream signaling molecules of this pathway,and the proteins that can directly affect the level of phosphorylation of AKT/mTOR in cancer cells.We also review the proteins that can co-regulate this pathway and its downstream pathways.Through this study,we hope to gain a deeper understanding of the regulatory mechanism of the AKT/mTOR pathway in cancer cells,in hopes of finding effective and harmless cancer treatment targets in the future.展开更多
BACKGROUND Pancreatic cancer(PC)is associated with some of the worst prognoses of all major cancers.Thymoquinone(TQ)has a long history in traditional medical practice and is known for its anti-cancer,anti-inflammatory...BACKGROUND Pancreatic cancer(PC)is associated with some of the worst prognoses of all major cancers.Thymoquinone(TQ)has a long history in traditional medical practice and is known for its anti-cancer,anti-inflammatory,anti-fibrosis and antioxidant pharmacological activities.Recent studies on hypoxia-inducible factor-1α(HIF-1α)and PC have shown that HIF-1αaffects the occurrence and development of PC in many aspects.In addition,TQ could inhibit the development of renal cancer by decreasing the expression of HIF-1α.Therefore,we speculate whether TQ affects HIF-1αexpression in PC cells and explore the mechanism.AIM To elucidate the effect of TQ in PC cells and the regulatory mechanism of HIF-1αexpression.METHODS Cell counting kit-8 assay,Transwell assay and flow cytometry were performed to detect the effects of TQ on the proliferative activity,migration and invasion ability and apoptosis of PANC-1 cells and normal pancreatic duct epithelial(hTERTHPNE)cells.Quantitative real-time polymerase chain reaction and western blot assay were performed to detect the expression of HIF-1αmRNA and protein in PC cells.The effects of TQ on the HIF-1αprotein initial expression pathway and ubiquitination degradation in PANC-1 cells were examined by western blot assay and co-immunoprecipitation.RESULTS TQ significantly inhibited proliferative activity,migration,and invasion ability and promoted apoptosis of PANC-1 cells;however,no significant effects on hTERT-HPNE cells were observed.TQ significantly reduced the mRNA and protein expression levels of HIF-1αin PANC-1,AsPC-1,and BxPC-3 cells.TQ significantly inhibited the expression of the HIF-1αinitial expression pathway(PI3K/AKT/mTOR)related proteins,and promoted the ubiquitination degradation of the HIF-1αprotein in PANC-1 cells.TQ had no effect on the hydroxylation and von Hippel Lindau protein mediated ubiquitination degradation of the HIF-1αprotein but affected the stability of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90,thus promoting its ubiquitination degradation.CONCLUSION The regulatory mechanism of TQ on HIF-1αprotein expression in PC cells was mainly to promote the ubiquitination degradation of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90;Secondly,TQ reduced the initial expression of HIF-1αprotein by inhibiting the PI3K/AKT/mTOR pathway.展开更多
Background: Depression is a typical psychosomatic disease. Shuganheweitang (SGHWT) is a clinical formula that effectively treats depression. However, the potential mechanism used by SGHWT to ameliorate depression-like...Background: Depression is a typical psychosomatic disease. Shuganheweitang (SGHWT) is a clinical formula that effectively treats depression. However, the potential mechanism used by SGHWT to ameliorate depression-like behaviors is still unclear. This study investigated the effects of SGHWT on metabolic change in the liver and hypothalamus with signaling pathways involved in chronic unpredictable mild stress (CUMS)-induced depression in rats to explore the mechanism of the anti-depressive effect. Methods: A total of 52 rats were used to create a model of depression by CUMS combined with solitary rearing for 6 weeks. Open field test (OFT), sucrose preference test (SPT), forced swim test (FST), and body weight (BW) were performed to analyze the pharmacodynamic effects of SGHWT. H&E staining, Nissl staining, immunofluorescence, immunohistochemistry, and western blot were used to evaluate the mechanism of action. Untargeted metabolomics techniques by ultra-performance liquid chromatography-quantitative time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) were used to analyze all the metabolic differences in the liver and hypothalamus. Results: SGHWT improved CUMS-induced depression-like behaviors in vivo. SGHWT reduced hepatic c-Fos protein expression and increased hypothalamic c-Fos protein expression. Moreover, p-PI3K, p-AKT473, p-AKT308, and p-mTOR protein expressions were significantly downregulated in the liver and hypothalamus of CUMS rats. Notably, these alterations were reversed by the SGHWT administration. Furthermore, the metabolomic analysis identified 15 and 5 key differential SPT-associated metabolites in the liver and hypothalamus, respectively. Conclusion: This study suggests that SGHWT ameliorates chronic unpredictable mild stress-induced depression-like behaviors, by the involvement of amino acids, glycerophospholipids, energy metabolism, and the PI3K/AKT/mTOR pathway. Highlights: 1) Shuganheweitang was derived from the TCM herbal formula Sinisan. 2) SGHWT treatment reverses depression-like behaviors in CUMS-induced rats. 3) The mechanism of SGHWT on depression by the liver and hypothalamus metabolomics. 4) SGHWT regulates amino acids, glycerophospholipids, and energy metabolism. 5) SGHWT exerts antidepressant effects through the PI3K/AKT/mTOR pathway.展开更多
Objective:To study the mechanism of action of Yanghe Huayan Decoction on endocrine-resistant cells(MCF-7R/mTOR cells)with low expression of mTOR;Methods:CCK-8 assay and cell clone assay were used to detect cell prolif...Objective:To study the mechanism of action of Yanghe Huayan Decoction on endocrine-resistant cells(MCF-7R/mTOR cells)with low expression of mTOR;Methods:CCK-8 assay and cell clone assay were used to detect cell proliferation and clonal ability,and flow cytometry was used to detect cell apoptosis.The changes of cytokines in ER-PI3K/Akt/mTOR signaling pathway were analyzed by blot and QPCR.Results:Yanghe Huayan Decoction could significantly affect the proliferation(p<0.05)and cloning ability(p<0.05)of MCF-7R/mTOR cells,promote cell apoptosis(p<0.01),and downregulate the expression of ER,PI3K,AKT,mTOR and p-mTOR(p<0.05).Conclusion:Yanghe Huayan Decoction can regulate the ER-PI3K/Akt/mTOR signaling pathway with multiple targets,and the use of combined mTOR inhibitors can regulate the ER-PI3K/Akt/mTOR signaling pathway more significantly.展开更多
Objective:To study the effect and mechanism of icariin on the migration,proliferation and apoptosis of mouse melanoma B16 cells.Methods:Mouse melanoma B16 cells were treated with icariin at different concentrations(0,...Objective:To study the effect and mechanism of icariin on the migration,proliferation and apoptosis of mouse melanoma B16 cells.Methods:Mouse melanoma B16 cells were treated with icariin at different concentrations(0,10,20,50μmol/L)for 24 hours.Cell proliferation,morphology,apoptosis and migration ability were detected,and the expression of PI3K/AKT/mTOR pathway related proteins was detected by Western blot assay.Results:After treatment with icariin,the inhibition rate and apoptosis rate of mouse melanoma B16 cells increased significantly with the increase of administration concentration(P<0.05).Hoechst 33258 staining showed that the cells in the blank control group(0μmol/L)were uniformly stained and the color was lighter,while the cells in the experimental group containing icariin were thicker in color.The higher the concentration of the icariin,the more obvious the degree of chromatin aggregation.The scratch healing rate of B16 cells and the cell count on the bottom of Transwell membrane decreased significantly with the increase of icariin concentration(P<0.05).The results of protein detection showed that with the increase of administration concentration,the expression of MMP-9,MMP-2 and mTOR decreased significantly,while the ratio of PI3K/pPI3K and AKT/pAKT increased significantly,and there was significant difference between the groups(P<0.05).Conclusions:Icariin can effectively inhibit the expression of PI3K/AKT/mTOR pathway related proteins in mouse melanoma B16 cells,thus inducing apoptosis of tumor cells,inhibiting cell migration and finally exerting antitumor effect.展开更多
Objective:To observe the effect of Taohong Siyu Decoction on the coagulation function and the signaling pathway of PI3K(phosphatidylinositol 3-kinase)/AKT(protein kinase B)/mTOR(mammalian target of rapamycin)after fem...Objective:To observe the effect of Taohong Siyu Decoction on the coagulation function and the signaling pathway of PI3K(phosphatidylinositol 3-kinase)/AKT(protein kinase B)/mTOR(mammalian target of rapamycin)after femoral artery anastomosis in rabbits.Methods:30 New Zealand white rabbits were divided into blank control group,model control group,papavine hydrochloride injection group and low,medium and high dose groups of Taohong Siwu decoction by random number table method,with 5 rabbits in each group.The rabbits in the model control group,papavine hydrochloride injection group and low,medium and high dose groups of Taohong Siwu decoction were treated with the femoral artery simple intermittent end-to-end suture model.After the successful modeling,the low,medium and high dose groups of Taohong Siwu decoction were given the Taohong Siwu decoction,while the model control group,the blank control group and papavine hydrochloride injection group were given the same amount of normal saline.APTT(activated partial thromboplastin time),FIB(fibrinogen)and PI3K/AKT/mTOR concentrations were measured in aural venous blood samples from six groups of rabbits 30min before operation and 1d,2D,3D and 7d after operation,respectively.Statistical analysis was conducted on the data of the six groups.Results:Compared with blank control group,APTT of model control group was significantly shortened 1d to 7d after operation(P<0.05),FIB values were significantly increased from 1d to 7d after operation(P<0.05);Compared with model control group,APTT in Taohong Siwu decoction low-dose,medium-dose and high-dose groups were significantly prolonged 1d to 7d after operation(P<0.05),FIB value of Taohong Siwu decoction medium and high dose groups decreased significantly from 1d to 7d after operation(P<0.05),the FIB value of Taohong Siwu decoction low-dose group was significantly decreased from 2d to 7d after surgery(P<0.05);Compared with papaverine hydrochloride injection group,APTT in Taohong Siwu decoction medium dose group was significantly prolonged 2d to 7d after surgery(P<0.05),APTT of Taohong Siwu decoction high-dose group was significantly prolonged on 1d to 7d after operation(P<0.05).FIB in Taohong Siwu decoction medium and high dose groups decreased significantly 1d to 7d after operation(P<0.05);Compared with the blank control group,the expression concentrations of PI3K,Akt and mTOR in serum of the model control group were significantly increased from 1d to 7d after surgery(P<0.05);Compared with the model control group,the expression levels of PI3K,Akt and mTOR in serum were significantly increased in the low dose group of Taohong Siwu decoction and Papaverine Hydrochloride Injection group on postoperative 7 days(P<0.05),Taohong Siwu decoction high-dose group was significantly increased from 1d to 7d after surgery(P<0.05),the expression concentrations of PI3K and Akt in Taohong Siwu decoction medium dose group were significantly increased from 2d to 7d after operation(P<0.05),mTOR expression levels were significantly increased from 3d to 7d after operation(P<0.05);Compared with papaverine hydrochloride injection group,the expression concentrations of PI3K,Akt and mTOR in serum of Taohong Siwu decoction medium dose group were significantly increased from 3d to 7d after operation(P<0.05),the expression concentrations of PI3K and mTOR in Taohong Siwu decoction high-dose group were significantly increased from 1d to 7d after operation(P<0.05),and the expression concentration of Akt increased significantly from 3d to 7d after operation(P<0.05).Conclusion:The Taohong Siwu decoction can improve the coagulation function of rabbit femoral artery anastomosis,prevent thrombosis,activate PI3K/Akt/mTOR signaling pathway,promote angiogenesis,and improve tissue ischemia after artery anastomosis.展开更多
Objective:To investigate the anti-atherosclerosis effect of chikusetsusaponinⅣ(CSⅣ)against high-fat diet-induced atherosclerosis in rats.Methods:A high-fat diet was used for the induction of atherosclerosis in rats,...Objective:To investigate the anti-atherosclerosis effect of chikusetsusaponinⅣ(CSⅣ)against high-fat diet-induced atherosclerosis in rats.Methods:A high-fat diet was used for the induction of atherosclerosis in rats,and the rats received oral CSⅣor atorvastatin.The body weight,organ weights,food intake,calorie intake,lipid parameters,3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA)/mevalonate ratio,collagen,free fatty acid,cardiac parameters,apolipoprotein(A and B),antioxidant parameters,inflammatory cytokines,and inflammatory parameters were assessed.The mRNA expressions of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6,IL-17,PI3K,AKT,and mTOR were estimated.Results:CSⅣsignificantly modulated food intake,body weight,organ weight(liver,kidney,and heart),and calories(P<0.05).Total cholesterol,triglycerides,very low-density lipoprotein cholesterol,low-density lipoprotein cholesterol,cardiovascular risk index-1,and cardiovascular risk index-2 were decreased,while high-density lipoprotein cholesterol and anti-atherogenic index were increased significantly in the CSⅣgroup(P<0.05).Besides,CSⅣsignificantly restored the level of HMG-CoA/mevalonate ratio,collagen,free fatty acid,cardiac parameters(creatinine kinase-MB,lactate dehydrogenase,cTnT,cTnI),apolipoprotein(apolipoprotein A and apolipoprotein B),antioxidant parameters(MDA,CAT,GPx,GSH,SOD),inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10),inflammatory parameters(COX-2,TGF-β,NF-κB),intercellular adhesion molecule-1,vascular cell adhesion molecule-1,and monocyte chemoattractant protein-1.CSⅣalso decreased the mRNA expression of IL-1β,TNF-α,IL-6,IL-17,PI3K,AKT,and mTOR.Conclusions:This study showed the anti-atherosclerosis effect of CSⅣagainst high-fat diet-induced atherosclerosis in rats via alteration of NF-κB/COX-2 and PI3K/AKT/mTOR signaling pathway.展开更多
Muscle atrophy can be induced by high doses or prolonged use of glucocorticoids.Kaempferol(Kae)is a naturally occurring flavonoid with a variety of biological activities and the effect of Kae on dexamethasone(Dex)indu...Muscle atrophy can be induced by high doses or prolonged use of glucocorticoids.Kaempferol(Kae)is a naturally occurring flavonoid with a variety of biological activities and the effect of Kae on dexamethasone(Dex)induced muscle atrophy in animals has not been elucidated.To explore this issue,the present experiments used a computationally assisted drug design scheme combining network pharmacology,molecular docking and in vivo experiments to investigate the mechanism of Kae against muscle atrophy.Network pharmacological analyses revealed 275 potential targets for Kae and 12294 potential targets for muscle atrophy,with a total of 228 crosstargets for Kae and muscle atrophy.GO and KEGG analyses were performed based on the protein-protein interaction(PPI)network of muscle atrophy and Kae component targets.The GO results showed that the biological processes were mainly related to the metabolic process of reactive oxygen species,and the response to oxidative stress;the cellular components were mainly focused on membrane microdomains,and membrane regions;the molecular functions mainly worked on phosphatase binding;and the KEGG pathway enrichment analyses identified the pathways of interaction between Kae and muscle atrophy.Finally,as verified by in vivo experiments,Kae may reduce the onset of muscle atrophy by activating the PI3K/AKT/m TOR/signalling pathway,inhibiting Foxo1/Foxo3 activity,and inhibiting downstream production of the ubiquitination 3 ligases Atrogin1 and Mu RF1;Kae also promotes the expression of NRF2/HO-1/KEAP1 signalling pathway,enhances muscle antioxidant capacity,inhibits the release of COX-2 and TNF-αinflammatory factors,and reduces the damage caused by oxidative stress and inflammatory factors to muscles.Therefore,there may be a synergistic effect of PI3K/AKT/m TOR and NRF2/HO-1/KEAP1 in Kae working together to prevent muscle atrophy.The binding energy and stability of Kae to potential targets were examined by molecular docking and molecular dynamics simulations,implying that Kae could be used for the prevention and treatment of muscle atrophy in patients.展开更多
Background:Interleukin-1 receptor-associated kinase 1(IRAK1),an active serine/threonine kinase,is an indispensable mediator of inflammatory responses and innate immunity.Emerging evidence has highlighted the oncogenic...Background:Interleukin-1 receptor-associated kinase 1(IRAK1),an active serine/threonine kinase,is an indispensable mediator of inflammatory responses and innate immunity.Emerging evidence has highlighted the oncogenic role of IRAK1 in tumors.However,the role of IRAK1 in gastric cancer(GC)progression remains unclear.Methods:IRAK1 expression levels in patients with GC at Peking Union Medical College Hospital were assessed by Western blotting,quantitative real-time polymerase chain reaction,and immunohistochemistry.To elucidate the role of IRAK1 in GC pathogenesis,we established GC cells with clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR-associated protein 9 (Cas9)mediated IRAK1 knockout and lentiviral vector-mediated IRAK1 overexpression and subsequently conducted in vitro and in vivo experiments.Additionally,we assessed the effect of IRAK1 expression levels in GC cells on the M2 polarization of tumor-associated macrophages(TAMs)via a Transwell coculture system.Functional assays were subsequently carried out to determine whether IRAK1 regulates the proliferation,invasion,and epithelial-mesenchymal transition(EMT)of GC cells through the induction of TAM M2 polarization and to clarify the associated regulatory mechanisms.Results:IRAK1 expression was progressively increased during GC progression.Clinicopathological feature analysis revealed that high IRAK1 expression predicted poor survival outcomes.In vitro and in vivo experiments revealed that IRAK1 was highly expressed in GC cells and facilitated the proliferation,migration,invasion,and EMT of GC cells via the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway.More interestingly,IRAK1 affected the malignant biological behavior of GC cells by inducing M2-like polarization of macrophages.Mechanistically,coculturing M0 macrophages with IRAK1-knockout GC cells suppressed interleukin(IL)-8 secretion,thereby inhibiting Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway activation in TAMs.Inactivation of the JAK2/STAT3 pathway suppressed the M2 polarization of TAMs,ultimately inhibiting GC progression.Conclusions:IRAK1 influences the malignant biological behavior of GC cells by activating the PI3K/AKT/mTOR pathway and inducing the M2-like polarization of macrophages via the IL-8/JAK2/STAT3 pathway in TAMs.Our findings provide a novel diagnostic biomarker and a promising therapeutic strategy for GC.展开更多
文摘AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.
基金supported by the National Nature Science Foundation of China,No.81471308(to JL)the Innovative Leading Talents of Liaoning Province,No.XLYC1902031(to JL)+2 种基金Science and Technology Projects in Liaoning Province,No.2022-BS-238(to CH)Young Top Talents of Liaoning Province,No.XLYC1907009(to LW)Dalian Science and Technology Innovation Fund,No.2018J11CY025(to JL)。
文摘Human neural stem cell-derived extracellular vesicles exhibit analogous functions to their parental cells,and can thus be used as substitutes for stem cells in stem cell therapy,thereby mitigating the risks of stem cell therapy and advancing the frontiers of stem cell-derived treatments.This lays a foundation for the development of potentially potent new treatment modalities for ischemic stroke.However,the precise mechanisms underlying the efficacy and safety of human neural stem cell-derived extracellular vesicles remain unclear,presenting challenges for clinical translation.To promote the translation of therapy based on human neural stem cell-derived extracellular vesicles from the bench to the bedside,we conducted a comprehensive preclinical study to evaluate the efficacy and safety of human neural stem cell-derived extracellular vesicles in the treatment of ischemic stroke.We found that administration of human neural stem cell-derived extracellular vesicles to an ischemic stroke rat model reduced the volume of cerebral infarction and promoted functional recovery by alleviating neuronal apoptosis.The human neural stem cell-derived extracellular vesicles reduced neuronal apoptosis by enhancing phosphorylation of phosphoinositide 3-kinase,mammalian target of rapamycin,and protein kinase B,and these effects were reversed by treatment with a phosphoinositide 3-kinase inhibitor.These findings suggest that human neural stem cell-derived extracellular vesicles play a neuroprotective role in ischemic stroke through activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway.Finally,we showed that human neural stem cell-derived extracellular vesicles have a good in vivo safety profile.Therefore,human neural stem cell-derived extracellular vesicles are a promising potential agent for the treatment of ischemic stroke.
文摘The published article titled“Knockdown of Rap1b Enhances Apoptosis and Autophagy in Gastric Cancer Cells via the PI3K/Akt/mTOR Pathway”has been retracted from Oncology Research,Vol.24,No.5,2016,pp.287–293.DOI:10.3727/096504016X14648701447779 URL:https://www.techscience.com/or/v24n5/56977 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases.
文摘Hepatocellular carcinoma(HCC)is the most common primary liver tumor and the third leading cause of cancer-related mortality globally.The phosphatidylinositol-3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway is critically involved in HCC pathogenesis,stimulating uncontrolled cell proliferation,survival,and tumor progression.The overactivation of this pathway is strongly linked to poor prognosis,making it a crucial target for therapeutic intervention.The oncogenic roles of PI3K/AKT/mTOR components in HCC have been highlighted,noting that class I PI3K deregulation,phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA)upregulation,and mTOR overexpression could be associated with poor HCC outcomes.To the best of our knowledge,this is the first time that the clinical trials investigating PI3K/AKT/mTOR inhibitors in HCC are analyzed.Accordingly,there is a predominance of mTOR inhibitors,with everolimus being the most frequently utilized drug.However,only 10%of studies advanced to phase III or IV,predominantly involving mTOR inhibitors.Challenges such as adverse events like hyperglycemia and bone marrow suppression,as well as the emergence of treatment resistance,have hindered the success of these therapies.Combination therapies,particularly those involving mitogen-activated protein kinase kinase(MEK)inhibitors,chemotherapy,immune checkpoint inhibitors,and vascular endothelial growth factor(VEGF)inhibitors,have shown promise in overcoming these challenges.Recent advances in nanotechnology offer the potential for improving drug delivery and reducing toxicity.
基金This work was supported by the National Natural Science Foundation of China(Grant Numbers 31971106,BWS21L013,21WS09002,JK20211A010213).
文摘Background:Oral squamous cell carcinoma(OSCC)is a common malignant tumor.Recently,Laminin Gamma 2(LAMC2)has been shown to be abnormally expressed in OSCC;however,how LAMC2 signaling contributes to the occurrence and development of OSCC and the role of autophagy in OSCC has not been fully explored.This study aimed to analyze the role and mechanism of LAMC2 signaling in OSCC and the involvement of autophagy in OSCC.Methods:To explore the mechanism by which LAMC2 is highly expressed in OSCC,we used small interfering RNA(siRNA)to knock down LAMC2 to further observe the changes in the signaling pathway.Furthermore,we used cell proliferation assays,Transwell invasion assays,and wound-healing assays to observe the changes in OSCC proliferation,invasion,and metastasis.RFP-LC3 was used to detect the level of autophagy intensity.A cell line-derived xenograft(CDX)model was used to detect the effect of LAMC2 on tumor growth in vivo.Results:This study found that the level of autophagy was correlated with the biological behavior of OSCC.The downregulation of LAMC2 activated autophagy and inhibited OSCC proliferation,invasion,and metastasis via inhibiting the PI3K/AKT/mTOR pathway.Moreover,autophagy has a dual effect on OSCC,and the synergistic downregulation of LAMC2 and autophagy can inhibit OSCC metastasis,invasion,and proliferation via the PI3K/AKT/mTOR pathway.Conclusions:LAMC2 interacts with autophagy to regulate OSCC metastasis,invasion,and proliferation via the PI3K/AKT/mTOR pathway.LAMC2 down-regulation can synergistically modulate autophagy to inhibit OSCC migration,invasion,and proliferation.
基金supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF)funded by the Ministry of Education,Science and Technology (NRF2020R1A2C1014798 to E-K Kim)。
文摘We evaluated the effect of isoquercetin(quercetin-O-3-glucoside-quercetin,IQ)as a functional component of Abeliophyllum disistichum Nakai ethanol extract(ADLE)on prostate cell proliferation and apoptosis and its effects on the IGF-1/PI3K/Akt/mTOR pathway in benign prostatic hyperplasia(BPH).Metabolites in ADLE were analyzed using UHPLC-qTOF-MS and HPLC.IQ was orally administered(1 or 10 mg/kg)to a testosterone propionate-induced BPH rat model,and its effects on the prostate weight were evaluated.The effect of IQ on androgen receptor(AR)signaling was analyzed in LNCaP cells.Whether IGF-1 and IQ affect the IGF-1/PI3K/Akt/mTOR pathway in BPH-1 cells was also examined.The metabolites in ADLE were identified and quantified,which confirmed that ADLE contained abundant IQ(20.88 mg/g).IQ significantly reduced the prostate size in a concentration-dependent manner in a BPH rat model,and significantly decreased the expression of AR signaling factors in the rat prostate tissue and LNCaP cells in a concentration-dependent manner.IQ also inhibited the PI3K/AKT/mTOR pathway activated by IGF-1 treatment in BPH-1 cells.In BPH-1 cells,IQ led to G0/G1 arrest and suppressed the expression of proliferation factors while inducing apoptosis.Thus,IQ shows potential for use as a pharmaceutical and nutraceutical for BPH.
基金supported by the National Key R&D Program of China(No.2018YFC0311206)the China Postdoctoral Science Foundation(No.2018M642706)the Postdoctoral Innovation Program of Shandong Province.
文摘A kind of triterpene glycosides echinoside A(EA)was extracted from sea cucumber Pearsonothuria graeffei,and its yield was about 0.78%.The purity of EA was 99.0%,and its molecular weight was 1206 Da.EA was a linear tetrasaccharide attached to a pentacyclic triterpene aglycon.It inhibited the growth of MDA-MB-231 cells in vitro.The antitumor effect was related to elevate ROS level,decrease mitochondrial membrane potential,enhance caspase-3 expression,induce cells apoptosis and arrest cell cycle at G2/M phase.EA also dose-dependently suppressed the expressions of phophorylation proteins p-PI3K,p-Akt,and p-mTOR as analyzed by western blotting.These results suggested that EA caused MDA-MB-231 cells apoptosis via intrinsic mitochondrial and PI3K/Akt/mTOR pathway.EA can be a potential anti-breast cancer agent to enhance the clinical efficacy.
基金National Natural Science Foundation of China(81473617)Natural Science Foundation of Hunan Province(2020JJ4066)+1 种基金Scientific Research Project of Hunan Education Department(18A266)Hunan Graduate Scientific Research Innovation Project(QL20210173)。
文摘Objective To investigate the effects of Niuhuang(Bovis Calculus,BC)and Shexiang(Moschus)(BC-Moschus)on human hepatocellular carcinoma(HCC)cells SMMC-7721 and a nude mouse model of subcutaneous xenografts,and to explore its anti-HCC mechanism.Methods The BC-Moschus combination was applied to two liver cancer models in vivo and in vitro.SMMC-7721 was divided into the BC-Moschus group and the control group,and different doses(rude drug dosage 0.625,1.25,2.5,and 5 mg/m L)of BC-Moschus extract were used for the intervention.The proliferation ability of HCC cells was detected using the Cell Counting Kit-8(CCK-8)assay,and the migration ability was detected by a wound healing assay.A subcutaneous xenograft model was prepared using nude mice with human HCC.Specific pathogen-free-grade BALB/c nude mice(5-week-old)were randomly divided into the following groups(n=6 per group):control(0.9%physiological saline 0.2 m L/d),BC-Moschus[BC 45.5 mg/(kg·d)+Moschus 13 mg/(kg·d)],and cisplatin(DDP,intraperitoneal injection5 mg/kg per week)groups.All groups were administered for 14 d.The volume and mass of the subcutaneous xenografts in nude mice were observed.The expression levels of phosphatidylinositol-3 kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway,apoptosis-associated factor p70 S6 Kinase(S6K),Bax,Bcl-2,caspase-3,and caspase-9 in nude mice subcutaneous xenografts were measured by real-time quantitative PCR(RT-qPCR)and Western blot.Terminal Deoxynucleotidy Transferase-Mediated d UTP NickEnd Labeling(TUNEL)was used for quantitative analysis of apoptotic cells.Results The CCK-8 assay demonstrated that the BC-Moschus combination inhibited HCC cell proliferation in a superior manner to the use of BC and Moschus alone,and the inhibition effect was dose-and time-dependent(P<0.01).The wound healing assay showed that the BC-Moschus combination inhibited HCC cell migration(P<0.01).In the subcutaneous xenograft model of nude mice with human HCC,we found that the tumor volume and weight of the BC-Moschus group were lower than those of the control group(P<0.01).The levels of the PI3K/AKT/m TOR signaling pathway and S6K protein in the BC-Moschus and DDP groups were significantly decreased(P<0.01).The expression level of the anti-apoptotic gene Bcl-2 was downregulated(P<0.05),and the expression of the pro-apoptotic gene Baxand apoptosis-related factors caspase-3 and caspase-9 were significantly upregulated(P<0.01).The TUNEL assays further confirmed that the combination of the BC-Moschuas could promote HCC(P<0.01).Conclusion The BC-Moschus combination inhibited the proliferation and migration ability of HCC cells SMMC-7721 and effectively inhibited the growth of subcutaneous xenografts in nude mice.The mechanism may be closely related to the downregulation of the PI3K/AKT/mTOR pathway,regulation of apoptosis-related protein caspase-3,caspase-9,Bcl-2,and Bax expression,and promotion of apoptosis.
基金This work was supported by the National Natural Science Foundation of China(No.82004027)the Fundamental Research Funds for the Central Universities(JUSRPI1961).
文摘Catechins have been proven to exert antitumor effects in different kinds of cancers.However,the underlying mechanisms have not been completely clarified yet.This study aimed to assess the effects and mechanisms of(-)-epigallocatechin-3-gallate(EGCG)and(-)-epicatechin-3-gallate(ECG)on human melanoma skin A375 cells.Results showed that EGCG and ECG inhibited the proliferation of A375 cells and ECG showed better inhibitory effect.Flow cytometry analysis had shown that EGCG and ECG induced apoptosis and led to cell cycle arrest.EGCG and ECG decreased Bcl-2 expression and upregulated Caspase-3 protein level,indicating the development of apoptosis.Furthermore,EGCG and ECG could decreased mitochondrial membrane potential of A375 cells.In addition,the expression of Beclin-1,LC3 and Sirt3 were downregulated at protein levels,which known to be associated with autophagy.After autophagy was increased by rapamycin,the apoptotic trend was not change,indicating that apoptosis and autophagy are independent.Mechanistically,EGCG and ECG treatments decreased phosphorylated-AMPK(p-AMPK)and increased the ratios of P-PI3K,p-AKT and p-mTOR in melanoma cells.Conclusively,EGCG and ECG induced apoptosis via mitochondrial signaling pathway,downregulated autophagy through modulating the AMPK/mTOR and PI3K/AKT/mTOR signaling pathway.It indicated that EGCG and ECG may be utilized in human melanoma treatment.
基金supported by the National Key R&D Program of China (2016YFD0400505)the National Natural Science Foundation of China (81600126)+1 种基金the Tianjin Municipal Education Commission (TD13-5013)the Key Research and Development Program of Shandong Province (2019YYSP011) the Tianjin Graduate Research Innovation Project (2020YJSB132)。
文摘Monascus vinegar(MV) is a typical fermented food with various health-promoting effects. This study aimed to evaluate the role of MV in alleviating high-fat-diet-induced inflammation in rats with hyperlipidemia and elucidate the possible regulatory mechanisms. In the study, serum lipid profiles, liver pathology and liver inflammatory cytokines were analyzed in hyperlipidemia rats with MV(0.5 mL/kg mb, 2 mL/kg mb). Results showed that the administration of MV alleviated dyslipidemia by decreasing the serum and liver levels of triglyceride and total cholesterol. Increase in hepatic lipase and carnitine palmitoyl transferase 1(CPT-1)levels and decrease in hepatocyte steatosis, nephritis, and intestinal tissue injury in the HD group showed that high-dose MV can significantly suppress hepatic lipid accumulation and steatosis. In addition, compared with the model(MOD) group, the HD group showed significantly down-regulated the level of serum or hepatic alanine aminotransferase(ALT), aspartate aminotransferase(AST), CPT-1, interleukin(IL)-2, IL-6, IL-12,and tumor necrosis factor α(TNF-α). Moreover, the HD group showed repressed hepatic nuclear factor κB(NF-κB) pathway and inactivated phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway mitigated liver inflammation. Similar results were obtained from cell experiments. Collectively, these findings revealed that MV might attenuate high-fat-diet-induced inflammation by inhibiting the NF-κB and PI3K/Akt/mTOR pathways.
基金supported by the National Natural Science Foundation of China(Nos.32102786,32270555).
文摘As a pathway that plays a role in nutrient absorption,anabolic response,cell growth and survival,the important role of AKT/mTOR in tumorigenesis has also come to light.For cancer patients,most deaths are caused by the growth of metastatic tumors outside the primary focus.Therefore,migration and invasion in the late stage of tumor progression are the main unresolved issues in the study of tumor pathogenesis,and AKT/mTOR has been found to participate in the migration and invasion of cancer cells,which means that the study of this pathway may contribute to a solution for the problem.Because of its extensive and complex functions in the organism,this pathway can be regulated by a variety of different signals in the body,and then realize its function through different downstream signal molecules.This article reviews the proteins that can indirectly affect this pathway by regulating the common upstream signaling molecules of this pathway,and the proteins that can directly affect the level of phosphorylation of AKT/mTOR in cancer cells.We also review the proteins that can co-regulate this pathway and its downstream pathways.Through this study,we hope to gain a deeper understanding of the regulatory mechanism of the AKT/mTOR pathway in cancer cells,in hopes of finding effective and harmless cancer treatment targets in the future.
基金Supported by Health Commission of Qinghai Province,No.2021-wjzdx-18.
文摘BACKGROUND Pancreatic cancer(PC)is associated with some of the worst prognoses of all major cancers.Thymoquinone(TQ)has a long history in traditional medical practice and is known for its anti-cancer,anti-inflammatory,anti-fibrosis and antioxidant pharmacological activities.Recent studies on hypoxia-inducible factor-1α(HIF-1α)and PC have shown that HIF-1αaffects the occurrence and development of PC in many aspects.In addition,TQ could inhibit the development of renal cancer by decreasing the expression of HIF-1α.Therefore,we speculate whether TQ affects HIF-1αexpression in PC cells and explore the mechanism.AIM To elucidate the effect of TQ in PC cells and the regulatory mechanism of HIF-1αexpression.METHODS Cell counting kit-8 assay,Transwell assay and flow cytometry were performed to detect the effects of TQ on the proliferative activity,migration and invasion ability and apoptosis of PANC-1 cells and normal pancreatic duct epithelial(hTERTHPNE)cells.Quantitative real-time polymerase chain reaction and western blot assay were performed to detect the expression of HIF-1αmRNA and protein in PC cells.The effects of TQ on the HIF-1αprotein initial expression pathway and ubiquitination degradation in PANC-1 cells were examined by western blot assay and co-immunoprecipitation.RESULTS TQ significantly inhibited proliferative activity,migration,and invasion ability and promoted apoptosis of PANC-1 cells;however,no significant effects on hTERT-HPNE cells were observed.TQ significantly reduced the mRNA and protein expression levels of HIF-1αin PANC-1,AsPC-1,and BxPC-3 cells.TQ significantly inhibited the expression of the HIF-1αinitial expression pathway(PI3K/AKT/mTOR)related proteins,and promoted the ubiquitination degradation of the HIF-1αprotein in PANC-1 cells.TQ had no effect on the hydroxylation and von Hippel Lindau protein mediated ubiquitination degradation of the HIF-1αprotein but affected the stability of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90,thus promoting its ubiquitination degradation.CONCLUSION The regulatory mechanism of TQ on HIF-1αprotein expression in PC cells was mainly to promote the ubiquitination degradation of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90;Secondly,TQ reduced the initial expression of HIF-1αprotein by inhibiting the PI3K/AKT/mTOR pathway.
文摘Background: Depression is a typical psychosomatic disease. Shuganheweitang (SGHWT) is a clinical formula that effectively treats depression. However, the potential mechanism used by SGHWT to ameliorate depression-like behaviors is still unclear. This study investigated the effects of SGHWT on metabolic change in the liver and hypothalamus with signaling pathways involved in chronic unpredictable mild stress (CUMS)-induced depression in rats to explore the mechanism of the anti-depressive effect. Methods: A total of 52 rats were used to create a model of depression by CUMS combined with solitary rearing for 6 weeks. Open field test (OFT), sucrose preference test (SPT), forced swim test (FST), and body weight (BW) were performed to analyze the pharmacodynamic effects of SGHWT. H&E staining, Nissl staining, immunofluorescence, immunohistochemistry, and western blot were used to evaluate the mechanism of action. Untargeted metabolomics techniques by ultra-performance liquid chromatography-quantitative time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) were used to analyze all the metabolic differences in the liver and hypothalamus. Results: SGHWT improved CUMS-induced depression-like behaviors in vivo. SGHWT reduced hepatic c-Fos protein expression and increased hypothalamic c-Fos protein expression. Moreover, p-PI3K, p-AKT473, p-AKT308, and p-mTOR protein expressions were significantly downregulated in the liver and hypothalamus of CUMS rats. Notably, these alterations were reversed by the SGHWT administration. Furthermore, the metabolomic analysis identified 15 and 5 key differential SPT-associated metabolites in the liver and hypothalamus, respectively. Conclusion: This study suggests that SGHWT ameliorates chronic unpredictable mild stress-induced depression-like behaviors, by the involvement of amino acids, glycerophospholipids, energy metabolism, and the PI3K/AKT/mTOR pathway. Highlights: 1) Shuganheweitang was derived from the TCM herbal formula Sinisan. 2) SGHWT treatment reverses depression-like behaviors in CUMS-induced rats. 3) The mechanism of SGHWT on depression by the liver and hypothalamus metabolomics. 4) SGHWT regulates amino acids, glycerophospholipids, and energy metabolism. 5) SGHWT exerts antidepressant effects through the PI3K/AKT/mTOR pathway.
基金National Natural Science Foundation of China(No.81573989)
文摘Objective:To study the mechanism of action of Yanghe Huayan Decoction on endocrine-resistant cells(MCF-7R/mTOR cells)with low expression of mTOR;Methods:CCK-8 assay and cell clone assay were used to detect cell proliferation and clonal ability,and flow cytometry was used to detect cell apoptosis.The changes of cytokines in ER-PI3K/Akt/mTOR signaling pathway were analyzed by blot and QPCR.Results:Yanghe Huayan Decoction could significantly affect the proliferation(p<0.05)and cloning ability(p<0.05)of MCF-7R/mTOR cells,promote cell apoptosis(p<0.01),and downregulate the expression of ER,PI3K,AKT,mTOR and p-mTOR(p<0.05).Conclusion:Yanghe Huayan Decoction can regulate the ER-PI3K/Akt/mTOR signaling pathway with multiple targets,and the use of combined mTOR inhibitors can regulate the ER-PI3K/Akt/mTOR signaling pathway more significantly.
基金Education project in Sichuan provincial department(NO.14ZB0196).
文摘Objective:To study the effect and mechanism of icariin on the migration,proliferation and apoptosis of mouse melanoma B16 cells.Methods:Mouse melanoma B16 cells were treated with icariin at different concentrations(0,10,20,50μmol/L)for 24 hours.Cell proliferation,morphology,apoptosis and migration ability were detected,and the expression of PI3K/AKT/mTOR pathway related proteins was detected by Western blot assay.Results:After treatment with icariin,the inhibition rate and apoptosis rate of mouse melanoma B16 cells increased significantly with the increase of administration concentration(P<0.05).Hoechst 33258 staining showed that the cells in the blank control group(0μmol/L)were uniformly stained and the color was lighter,while the cells in the experimental group containing icariin were thicker in color.The higher the concentration of the icariin,the more obvious the degree of chromatin aggregation.The scratch healing rate of B16 cells and the cell count on the bottom of Transwell membrane decreased significantly with the increase of icariin concentration(P<0.05).The results of protein detection showed that with the increase of administration concentration,the expression of MMP-9,MMP-2 and mTOR decreased significantly,while the ratio of PI3K/pPI3K and AKT/pAKT increased significantly,and there was significant difference between the groups(P<0.05).Conclusions:Icariin can effectively inhibit the expression of PI3K/AKT/mTOR pathway related proteins in mouse melanoma B16 cells,thus inducing apoptosis of tumor cells,inhibiting cell migration and finally exerting antitumor effect.
文摘Objective:To observe the effect of Taohong Siyu Decoction on the coagulation function and the signaling pathway of PI3K(phosphatidylinositol 3-kinase)/AKT(protein kinase B)/mTOR(mammalian target of rapamycin)after femoral artery anastomosis in rabbits.Methods:30 New Zealand white rabbits were divided into blank control group,model control group,papavine hydrochloride injection group and low,medium and high dose groups of Taohong Siwu decoction by random number table method,with 5 rabbits in each group.The rabbits in the model control group,papavine hydrochloride injection group and low,medium and high dose groups of Taohong Siwu decoction were treated with the femoral artery simple intermittent end-to-end suture model.After the successful modeling,the low,medium and high dose groups of Taohong Siwu decoction were given the Taohong Siwu decoction,while the model control group,the blank control group and papavine hydrochloride injection group were given the same amount of normal saline.APTT(activated partial thromboplastin time),FIB(fibrinogen)and PI3K/AKT/mTOR concentrations were measured in aural venous blood samples from six groups of rabbits 30min before operation and 1d,2D,3D and 7d after operation,respectively.Statistical analysis was conducted on the data of the six groups.Results:Compared with blank control group,APTT of model control group was significantly shortened 1d to 7d after operation(P<0.05),FIB values were significantly increased from 1d to 7d after operation(P<0.05);Compared with model control group,APTT in Taohong Siwu decoction low-dose,medium-dose and high-dose groups were significantly prolonged 1d to 7d after operation(P<0.05),FIB value of Taohong Siwu decoction medium and high dose groups decreased significantly from 1d to 7d after operation(P<0.05),the FIB value of Taohong Siwu decoction low-dose group was significantly decreased from 2d to 7d after surgery(P<0.05);Compared with papaverine hydrochloride injection group,APTT in Taohong Siwu decoction medium dose group was significantly prolonged 2d to 7d after surgery(P<0.05),APTT of Taohong Siwu decoction high-dose group was significantly prolonged on 1d to 7d after operation(P<0.05).FIB in Taohong Siwu decoction medium and high dose groups decreased significantly 1d to 7d after operation(P<0.05);Compared with the blank control group,the expression concentrations of PI3K,Akt and mTOR in serum of the model control group were significantly increased from 1d to 7d after surgery(P<0.05);Compared with the model control group,the expression levels of PI3K,Akt and mTOR in serum were significantly increased in the low dose group of Taohong Siwu decoction and Papaverine Hydrochloride Injection group on postoperative 7 days(P<0.05),Taohong Siwu decoction high-dose group was significantly increased from 1d to 7d after surgery(P<0.05),the expression concentrations of PI3K and Akt in Taohong Siwu decoction medium dose group were significantly increased from 2d to 7d after operation(P<0.05),mTOR expression levels were significantly increased from 3d to 7d after operation(P<0.05);Compared with papaverine hydrochloride injection group,the expression concentrations of PI3K,Akt and mTOR in serum of Taohong Siwu decoction medium dose group were significantly increased from 3d to 7d after operation(P<0.05),the expression concentrations of PI3K and mTOR in Taohong Siwu decoction high-dose group were significantly increased from 1d to 7d after operation(P<0.05),and the expression concentration of Akt increased significantly from 3d to 7d after operation(P<0.05).Conclusion:The Taohong Siwu decoction can improve the coagulation function of rabbit femoral artery anastomosis,prevent thrombosis,activate PI3K/Akt/mTOR signaling pathway,promote angiogenesis,and improve tissue ischemia after artery anastomosis.
基金funded by the Yancheng Municipal Health Commission 2024 Medical Research Project(YK2024166).
文摘Objective:To investigate the anti-atherosclerosis effect of chikusetsusaponinⅣ(CSⅣ)against high-fat diet-induced atherosclerosis in rats.Methods:A high-fat diet was used for the induction of atherosclerosis in rats,and the rats received oral CSⅣor atorvastatin.The body weight,organ weights,food intake,calorie intake,lipid parameters,3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA)/mevalonate ratio,collagen,free fatty acid,cardiac parameters,apolipoprotein(A and B),antioxidant parameters,inflammatory cytokines,and inflammatory parameters were assessed.The mRNA expressions of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6,IL-17,PI3K,AKT,and mTOR were estimated.Results:CSⅣsignificantly modulated food intake,body weight,organ weight(liver,kidney,and heart),and calories(P<0.05).Total cholesterol,triglycerides,very low-density lipoprotein cholesterol,low-density lipoprotein cholesterol,cardiovascular risk index-1,and cardiovascular risk index-2 were decreased,while high-density lipoprotein cholesterol and anti-atherogenic index were increased significantly in the CSⅣgroup(P<0.05).Besides,CSⅣsignificantly restored the level of HMG-CoA/mevalonate ratio,collagen,free fatty acid,cardiac parameters(creatinine kinase-MB,lactate dehydrogenase,cTnT,cTnI),apolipoprotein(apolipoprotein A and apolipoprotein B),antioxidant parameters(MDA,CAT,GPx,GSH,SOD),inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10),inflammatory parameters(COX-2,TGF-β,NF-κB),intercellular adhesion molecule-1,vascular cell adhesion molecule-1,and monocyte chemoattractant protein-1.CSⅣalso decreased the mRNA expression of IL-1β,TNF-α,IL-6,IL-17,PI3K,AKT,and mTOR.Conclusions:This study showed the anti-atherosclerosis effect of CSⅣagainst high-fat diet-induced atherosclerosis in rats via alteration of NF-κB/COX-2 and PI3K/AKT/mTOR signaling pathway.
基金funded by Yunnan Youth Top-notch Talent Support Program(YNWR-QNBJ2018-173)Agricultural Joint project of Yunnan Provincial S&T Programs(202301BD070001-195)+2 种基金S&T project of Yunnan provincial finance(K212020001-01)supported by Yunnan Province Education Department’s Engineering Research Center of Eco-friendly Products from Yunnan Characteristic Edible FungiYunnan Province Yongsheng County Farmer Academician Technology service station.
文摘Muscle atrophy can be induced by high doses or prolonged use of glucocorticoids.Kaempferol(Kae)is a naturally occurring flavonoid with a variety of biological activities and the effect of Kae on dexamethasone(Dex)induced muscle atrophy in animals has not been elucidated.To explore this issue,the present experiments used a computationally assisted drug design scheme combining network pharmacology,molecular docking and in vivo experiments to investigate the mechanism of Kae against muscle atrophy.Network pharmacological analyses revealed 275 potential targets for Kae and 12294 potential targets for muscle atrophy,with a total of 228 crosstargets for Kae and muscle atrophy.GO and KEGG analyses were performed based on the protein-protein interaction(PPI)network of muscle atrophy and Kae component targets.The GO results showed that the biological processes were mainly related to the metabolic process of reactive oxygen species,and the response to oxidative stress;the cellular components were mainly focused on membrane microdomains,and membrane regions;the molecular functions mainly worked on phosphatase binding;and the KEGG pathway enrichment analyses identified the pathways of interaction between Kae and muscle atrophy.Finally,as verified by in vivo experiments,Kae may reduce the onset of muscle atrophy by activating the PI3K/AKT/m TOR/signalling pathway,inhibiting Foxo1/Foxo3 activity,and inhibiting downstream production of the ubiquitination 3 ligases Atrogin1 and Mu RF1;Kae also promotes the expression of NRF2/HO-1/KEAP1 signalling pathway,enhances muscle antioxidant capacity,inhibits the release of COX-2 and TNF-αinflammatory factors,and reduces the damage caused by oxidative stress and inflammatory factors to muscles.Therefore,there may be a synergistic effect of PI3K/AKT/m TOR and NRF2/HO-1/KEAP1 in Kae working together to prevent muscle atrophy.The binding energy and stability of Kae to potential targets were examined by molecular docking and molecular dynamics simulations,implying that Kae could be used for the prevention and treatment of muscle atrophy in patients.
文摘Background:Interleukin-1 receptor-associated kinase 1(IRAK1),an active serine/threonine kinase,is an indispensable mediator of inflammatory responses and innate immunity.Emerging evidence has highlighted the oncogenic role of IRAK1 in tumors.However,the role of IRAK1 in gastric cancer(GC)progression remains unclear.Methods:IRAK1 expression levels in patients with GC at Peking Union Medical College Hospital were assessed by Western blotting,quantitative real-time polymerase chain reaction,and immunohistochemistry.To elucidate the role of IRAK1 in GC pathogenesis,we established GC cells with clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR-associated protein 9 (Cas9)mediated IRAK1 knockout and lentiviral vector-mediated IRAK1 overexpression and subsequently conducted in vitro and in vivo experiments.Additionally,we assessed the effect of IRAK1 expression levels in GC cells on the M2 polarization of tumor-associated macrophages(TAMs)via a Transwell coculture system.Functional assays were subsequently carried out to determine whether IRAK1 regulates the proliferation,invasion,and epithelial-mesenchymal transition(EMT)of GC cells through the induction of TAM M2 polarization and to clarify the associated regulatory mechanisms.Results:IRAK1 expression was progressively increased during GC progression.Clinicopathological feature analysis revealed that high IRAK1 expression predicted poor survival outcomes.In vitro and in vivo experiments revealed that IRAK1 was highly expressed in GC cells and facilitated the proliferation,migration,invasion,and EMT of GC cells via the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway.More interestingly,IRAK1 affected the malignant biological behavior of GC cells by inducing M2-like polarization of macrophages.Mechanistically,coculturing M0 macrophages with IRAK1-knockout GC cells suppressed interleukin(IL)-8 secretion,thereby inhibiting Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway activation in TAMs.Inactivation of the JAK2/STAT3 pathway suppressed the M2 polarization of TAMs,ultimately inhibiting GC progression.Conclusions:IRAK1 influences the malignant biological behavior of GC cells by activating the PI3K/AKT/mTOR pathway and inducing the M2-like polarization of macrophages via the IL-8/JAK2/STAT3 pathway in TAMs.Our findings provide a novel diagnostic biomarker and a promising therapeutic strategy for GC.
