Fibroblasts support a broad range of essential organ functions via microarchitectural,biomechanical,and biochemical cues.Despite great advances in fluorescence,photoacoustic conversion,and Raman scattering over the pa...Fibroblasts support a broad range of essential organ functions via microarchitectural,biomechanical,and biochemical cues.Despite great advances in fluorescence,photoacoustic conversion,and Raman scattering over the past decades,their invasiveness and limited spatial resolution hinder the characterization of fibroblasts in a single cell.Here,taking mouse embryonic fibroblasts(MEFs)as an example,we propose a novel noninvasive approach to investigate the compositional distribution of MEFs at the single-cell scale via terahertz(THz)nanos⁃copy.Compared to the topological morphology,THz nano-imaging enables the component-based visualization of MEFs,such as the membrane,cytoplasm,nucleus,and extracellular vesicles(EVs).Notably,we demonstrate the real-space observation of the influence of rapamycin treatment on the increase of EVs in MEFs.Moreover,the line-cut and area-statistical analysis establishes the relationship between the topological morphology and the THz near-field amplitudes for different cellular components of MEFs.This work provides a new pathway to char⁃acterize the effects of pharmaceutical treatments,with potential applications in disease diagnosis and drug devel⁃opment.展开更多
Pancreatic ductal adenocarcinoma stands out as an exceptionally fatal cancer owing to the complexities associated with its treatment and diagnosis,leading to a notably low five-year survival rate.This study offers a d...Pancreatic ductal adenocarcinoma stands out as an exceptionally fatal cancer owing to the complexities associated with its treatment and diagnosis,leading to a notably low five-year survival rate.This study offers a detailed exploration of epidemiological trends in pancreatic cancer and key molecular drivers,such as mutations in CDKN2A,KRAS,SMAD4,and TP53,along with the influence of cancer-associated fibroblasts(CAFs)on disease progression.In particular,we focused on the pivotal roles of signaling pathways such as the transforming growth factor-βand Wnt/β-catenin pathways in the development of pancreatic cancer and investigated their application in emerging therapeutic strategies.This study provides new scientific perspectives on pancreatic cancer treatment,especially in the development of precision medicine and targeted therapeutic strategies,and demonstrates the importance of signaling pathway research in the development of effective therapeutic regimens.Future studies should explore the subtypes of CAFs and their specific roles in the tumor microenvironment to devise more effective therapeutic methods.展开更多
Fibroblast activation protein(FAP)is overexpressed in cancer-associated fibroblasts across various cancer types.Numerous radiolabeled FAP inhibitors(FAPIs)(Fig.S1A)currently under clinical investigation have shown rem...Fibroblast activation protein(FAP)is overexpressed in cancer-associated fibroblasts across various cancer types.Numerous radiolabeled FAP inhibitors(FAPIs)(Fig.S1A)currently under clinical investigation have shown remarkable potential in cancer theranostics.展开更多
Breast cancer is one of the most common malignancies worldwide and is a major cause of cancer-related mortality among women.Beyond tumor cells,the tumor microenvironment(TME)also plays an important role in cancer prog...Breast cancer is one of the most common malignancies worldwide and is a major cause of cancer-related mortality among women.Beyond tumor cells,the tumor microenvironment(TME)also plays an important role in cancer progression,therapy resistance,and metastasis.The TME is a complex ecosystem consisting of stromal and immune cells,extracellular matrix(ECM),and various signaling molecules that dynamically interact with tumor cells.Cancer-associated fibro-blasts remodel the ECM and secrete growth factors that promote tumor growth and invasion.Immune cells,such as tumor-associated macrophages,regulatory T cells,and myeloid-derived suppressor cells,often contribute to an immunosup-pressive environment that hinders anti-tumor immune responses.The ECM pro-vides structural support and acts as a reservoir for signaling molecules that in-fluence cancer cell behavior.These components evolve together with tumor cells,facilitating immune evasion,therapy resistance,and epithelial-to-mesenchymal transition,which promotes metastasis.Understanding these interactions is nece-ssary to develop novel therapeutic strategies that target both tumor and micro-environmental components.This minireview highlights the key stromal and immune elements within the breast cancer microenvironment,discussing their individual and collective roles in tumor progression and clinical outcomes,while emphasizing emerging therapeutic approaches aiming to reprogram the TME to improve treatment efficacy.展开更多
Pancreatic cancer is a highly aggressive malignancy with a poor prognosis and limited therapeutic options.The tumor microenvironment(TME),including cancer-associated fibroblasts(CAFs),plays a pivotal role in tumor pro...Pancreatic cancer is a highly aggressive malignancy with a poor prognosis and limited therapeutic options.The tumor microenvironment(TME),including cancer-associated fibroblasts(CAFs),plays a pivotal role in tumor progression and therapy resistance.Senescent CAFs,which exhibit a senescence-associated secretory phenotype(SASP),further exacerbate cancer growth through inflammatory cytokine secretion.This editorial highlights a study by Jiang et al,which investigates the potential of resveratrol,a natural polyphenolic compound,in targeting senescent CAFs to inhibit pancreatic cancer progression.The study demonstrates that resveratrol reduces senescent CAFs and downregulates SASP factors,thereby disrupting the pro-tumorigenic activities of these cells.Resveratrol’s ability to modulate the TME,induce apoptosis in pancreatic cancer cells,and inhibit metastasis underscores its potential as an adjunctive therapy.This research offers promising insights into novel strategies for improving therapeutic outcomes in pancreatic cancer by targeting the TME and senescent CAFs.展开更多
The resistance to cancer treatment is a major clinical obstacle,being strongly influenced by the tumor microen-vironment(TME).Cancer-associated fibroblasts(CAFs)are critical elements of the TME.CAFs are heterogeneous ...The resistance to cancer treatment is a major clinical obstacle,being strongly influenced by the tumor microen-vironment(TME).Cancer-associated fibroblasts(CAFs)are critical elements of the TME.CAFs are heterogeneous and are activated through diverse pathways.These CAFs engage in reciprocal interactions with tumor cells,driv-ing tumor progression and therapeutic resistance.In this review,we discuss the role of CAFs in the development of tumor resistance to chemotherapy,radiotherapy,targeted therapy,and immunotherapy.Besides,we sum-marize recent clinical trials in CAF-targeted therapies.The development of resistance involves physical barrier formation,metabolic reprogramming,exosome release,DNA repair,bypass pathway activation,multidrug resis-tance protein upregulation,and immune checkpoint inhibition.Challenges remain in addressing drug resistance despite the therapeutic potential of targeting CAFs:the cellular origins of CAFs need to be clarified,and their limited clinical applications need to be increased.Future studies should focus on elucidating the reasons for CAF heterogeneity,developing precise targeting strategies,and validating the clinical safety and efficacy of CAF-based therapies to overcome treatment resistance and improve patient outcomes.展开更多
Objectives:The present study investigated whether Tripartite Motif-Containing Protein 32(TRIM32)contributes to the aberrant activation of keloid fibroblasts(KFs)via glycolysis.Methods:The expression levels of TRIM32,p...Objectives:The present study investigated whether Tripartite Motif-Containing Protein 32(TRIM32)contributes to the aberrant activation of keloid fibroblasts(KFs)via glycolysis.Methods:The expression levels of TRIM32,pyruvate dehydrogenase kinase 1(PDK1),hexokinase 2(HK2),and glucose transporter 1(GLUT1)in normal human skin fibroblasts(NFs)and KFs were analyzed using RT-qPCR analyses and western blotting.Cellular proliferation,invasion,and migration were evaluated using Transwell,wound healing,5-ethynyl-2′-deoxyuridine(EdU),and cell counting kit-8(CCK-8)assays.The extracellular acidification rate(ECAR)was measured using the XF96 Extracellular Flux Analyzer.Glucose uptake and ATP production were measured using specific assay kits.The expression ofα-smooth muscle actin(α-SMA)was determined by immunofluorescence assays.The expression levels of collagen I,α-smooth muscle actin(α-SMA),fibronectin(FN),and components of the phosphoinositide-3-kinase/protein kinase B(PI3K/AKT)signaling pathway were quantified by western blotting.Results:The expression of TRIM32 and glycolysis-related proteins was significantly elevated in KFs compared to that in NFs.TRIM32 overex-pression enhanced the proliferation,invasion,and migration of KFs,as well as extracellular matrix(ECM)deposition,glucose uptake,and ATP production,while TRIM32 silencing produced the opposite effects.The glycolysis inhibitor,2-deoxy-glucose(2-DG),significantly suppressed the biological functions of KFs;however,TRIM32 overexpression effectively counteracted the inhibitory effects of 2-DG.TRIM32 activated the PI3K/AKT signaling pathway in KFs.The PI3K inhibitor LY294002 decreased cellular glycolysis,with TRIM32 overexpression mitigated these inhibitory effects.Conclusion:This study demonstrated that TRIM32 enhances the viability of KFs by regulating glycolytic activity,potentially mediated via the PI3K/AKT signaling pathway,thereby suggesting novel therapeutic approaches for the treatment of keloids.展开更多
Perineural invasion(PNI)by tumor cells is a key phenotype of highly-invasive oral squamous cell carcinoma(OSCC).Since Schwann cells(SCs)and fibroblasts maintain the physiological homeostasis of the peripheral nervous ...Perineural invasion(PNI)by tumor cells is a key phenotype of highly-invasive oral squamous cell carcinoma(OSCC).Since Schwann cells(SCs)and fibroblasts maintain the physiological homeostasis of the peripheral nervous system,and we have focused on cancer-associated fibroblasts(CAFs)for decades,it’s imperative to elucidate the impact of CAFs on SCs in PNI+OSCCs.We describe a disease progression-driven shift of PNI−towards PNI+during the progression of early-stage OSCC(31%,n=125)to late-stage OSCC(53%,n=97),characterized by abundant CAFs and nerve demyelination.CAFs inhibited SC proliferation/migration and reduced neurotrophic factors and myelin in vitro,and this involved up-regulated ER stress and decreased MAPK signals.Moreover,CAFs also aggravated the paralysis of the hind limb and PNI in vivo.Unexpectedly,leukemia inhibitory factor(LIF)was exclusively expressed on CAFs and up-regulated in metastatic OSCC.The LIF inhibitor EC330 restored CAF-induced SC inactivation.Thus,OSCC-derived CAFs inactivate SCs to aggravate nerve injury and PNI development.展开更多
Objectives:Recently,pre-/post-operative Local Estrogen Therapy(LET)has shown effectiveness in alleviating Pelvic Organ Prolapse(POP)symptoms in clinical therapy.However,there is a lack of scientific evidence to suppor...Objectives:Recently,pre-/post-operative Local Estrogen Therapy(LET)has shown effectiveness in alleviating Pelvic Organ Prolapse(POP)symptoms in clinical therapy.However,there is a lack of scientific evidence to support these claims.Therefore,we aimed to explore the anti-senescence effects and mechanisms of 17β-estradiol(E2)on POP-derived fibroblasts.Methods:The primary fibroblast cells were isolated and cultured fromthe surgical samples of postmenopausal women clinically diagnosed with pelvic organ prolapse(POP)at stages III-IV(quantified using the POP-Q system)and without any other treatment within 6 months.(n=12,age 50–75).Colorimetric Cell Counting Kit(CCK-8)assay and Senescence-Associated-β-Galactosidase(SA-β-Gal)staining were used to test the cell proliferative capacity and the senescence rate.Western blotting(WB)was used to detect the expression of Collagen Type I(COL-I),Collagen Type III(COL-III),Cyclin-dependent kinase 4 inhibitor A(p16INK4a),Cyclin-dependent kinase inhibitor 1A(p21),Tumor Protein 53(p53),Sirtuin 1(SIRT-1)and Microtubule-associated protein 1A/1B-light chain 3-I/II(LC3-I/II)protein.A transmission ElectronMicroscope(TEM)was used to observe the ultrastructure of fibroblasts.Results:The results showed that E2 significantly promoted the proliferation of fibroblasts derived from POP and reduced the staining rate of SA-β-Gal.It markedly enhanced the extracellular matrix proteins COL-I and COL-III,accompanied by inhibition of the senescent maker p16INK4a.Additionally,our results improved the cells’autophagy and metabolic activity.Additionally,our results indicate the anti-senescence mechanism of E2 through the mediated SIRT-1/p53/p21 axis pathway.Conclusion:We provide preliminary evidence for the anti-aging effects and mechanisms of E2 on POP,hoping to provide a theoretical basis for estrogen against POP senescence and guide the clinical application and local administration of estrogen in POP treatment.展开更多
Osteosarcoma(OS)is a prevalent primary bone malignancy with limited treatment options.Therefore,it is imperative to investigate and understand the mechanisms underlying OS pathogenesis.Cancer-associated fibroblasts(CA...Osteosarcoma(OS)is a prevalent primary bone malignancy with limited treatment options.Therefore,it is imperative to investigate and understand the mechanisms underlying OS pathogenesis.Cancer-associated fibroblasts(CAFs)are markedly abundant in tumor stromal cells and are essentially involved in the modulation of tumor occurrence and development.In recent years,CAFs have become a hotspot as researchers aim to elucidate CAF mechanisms that regulate tumor progression.However,most studies on CAFs are limited to a few common cancers,and their association with OS remains elusive.This review describes the role and current knowledge of CAFs in OS,focusing on their potential cellular origin,classification,and diverse functionality.It was found that CAFs influenced OS tumor cell signaling,proliferation,invasion,metastasis,epithelial-mesenchymal transition,stemness maintenance,angiogenesis,and the ability to modify immune system components.Furthermore,findings on other common cancers indicated that effective therapeutic strategies included the manipulation of CAF activation,targeting CAF-derived components,and depletion of CAFs by biomarkers.This review provides new insights and a theoretical basis for OS research.展开更多
AIM:To explore the effect of co-host non-coding RNA(ncRNA)MIR503HG/miR-503-5p on the angiogenesis of pterygium.METHODS:MIR503HG/miR-503-5p/fibroblast growth factor 2(FGF2)expression levels in pterygium tissues,control...AIM:To explore the effect of co-host non-coding RNA(ncRNA)MIR503HG/miR-503-5p on the angiogenesis of pterygium.METHODS:MIR503HG/miR-503-5p/fibroblast growth factor 2(FGF2)expression levels in pterygium tissues,control conjunctival tissues,and human pterygium fibroblasts(HPF)were examined by reverse transcription-polymerase chain reaction(qRT-PCR)and immunohistochemical methods.Effects of MIR503HG/miR-503-5p on low molecular weight FGF2(LWM FGF2),migration and angiogenesis of human retinal microvascular endothelial cells(HRMEC)were determined in an HPF and HRMEC co-culture model using Western blots,wound healing assay,Matrigel-based tube formation assay,and Transwell assay.RESULTS:MIR503HG/miR-503-5p/FGF2 pathway was actively increased in pterygium tissue and there was a negative correlation between the expression of the two ncRNAs.FGF2 expression level was positively correlated with MIR503HG and negatively correlated with miR-503-5p.Overexpressed MIR503HG/miR-503-5p did not affect the migration and angiogenesis of HRMECs cultured separately,but significantly affected migration and angiogenesis of HRMEC in HPF and HRMEC co-culture models.Western blotting revealed that MIR503HG/miR-503-5p overexpression significantly increased LMW FGF2 expression in HPF.CONCLUSION:MIR503HG/miR-503-5p inhibits HRMEC migration and angiogenic function by interfering with the interaction between HPF and endothelial cells via reducing LMW FGF2 in HPF.展开更多
[Objective] The aim of this study was to establish the in vitro culture system of chicken fibroblasts.[Method] Tissue explant method and enzymatic digestion method were used to separate and culture chicken skin fibrob...[Objective] The aim of this study was to establish the in vitro culture system of chicken fibroblasts.[Method] Tissue explant method and enzymatic digestion method were used to separate and culture chicken skin fibroblasts respectively.The rate of cell growth,cryopreservation and recovery were compared.[Result] The primary chicken fibroblasts prepared by enzymatic digestion grew faster and converged together to form monolayer on 5 d post preparation;the passage cells prepared by these 2 methods grew at similar speed and formed monolayer within 2-3 d;homogeneous fibroblasts could be obtained by trypsin digestion and repeated attachment for 3-4 passages;there were 75%-80% of cells survived after cryopreservation and recovery;the growth curves of embryonic fibroblasts and skin fibroblasts were all normal and the two kind of cells still retained the normal number of chromosomes even at the twelfth passage.[Conclusion] The feeder layer cells needed for establishing ES cell lines could be obtained by culturing chicken fibroblasts through both tissue explant method and enzymatic digestion method.This study provided a basis for the successful establishment of ES cell lines.展开更多
Recent studies have shown that fibrotic scar formation following cerebral ischemic injury has varying effects depending on the microenvironment.However,little is known about how fibrosis is induced and regulated after...Recent studies have shown that fibrotic scar formation following cerebral ischemic injury has varying effects depending on the microenvironment.However,little is known about how fibrosis is induced and regulated after cerebral ischemic injury.Sonic hedgehog signaling participates in fibrosis in the heart,liver,lung,and kidney.Whether Shh signaling modulates fibrotic scar formation after cerebral ischemic stroke and the underlying mechanisms are unclear.In this study,we found that Sonic Hedgehog expression was upregulated in patients with acute ischemic stroke and in a middle cerebral artery occlusion/reperfusion injury rat model.Both Sonic hedgehog and Mitofusin 2 showed increased expression in the middle cerebral artery occlusion rat model and in vitro fibrosis cell model induced by transforming growth factor-beta 1.Activation of the Sonic hedgehog signaling pathway enhanced the expression of phosphorylated Smad 3 and Mitofusin 2 proteins,promoted the formation of fibrotic scars,protected synapses or promoted synaptogenesis,alleviated neurological deficits following middle cerebral artery occlusion/reperfusion injury,reduced cell apoptosis,facilitated the transformation of meninges fibroblasts into myofibroblasts,and enhanced the proliferation and migration of meninges fibroblasts.The Smad3 phosphorylation inhibitor SIS3 reversed the effects induced by Sonic hedgehog signaling pathway activation.Bioinformatics analysis revealed significant correlations between Sonic hedgehog and Smad3,between Sonic hedgehog and Mitofusin 2,and between Smad3 and Mitofusin 2.These findings suggest that Sonic hedgehog signaling may influence Mitofusin 2 expression by regulating Smad3 phosphorylation,thereby modulating the formation of early fibrotic scars following cerebral ischemic stroke and affecting prognosis.The Sonic Hedgehog signaling pathway may serve as a new therapeutic target for stroke treatment.展开更多
AAV-PHP.eB is an artificial adeno-associated virus(AAV)that crosses the blood-brain barrier and targets neurons more efficiently than other AAVs when administered systematically.While AAV-PHP.eB has been used in vario...AAV-PHP.eB is an artificial adeno-associated virus(AAV)that crosses the blood-brain barrier and targets neurons more efficiently than other AAVs when administered systematically.While AAV-PHP.eB has been used in various disease models,its cellular tropism in cerebrovascular diseases remains unclear.In the present study,we aimed to elucidate the tropism of AAV-PHP.eB for different cell types in the brain in a mouse model of ischemic stroke and evaluate its effectiveness in mediating basic fibroblast growth factor(bFGF)gene therapy.Mice were injected intravenously with AAV-PHP.eB either 14 days prior to(pre-stroke)or 1 day following(post-stroke)transient middle cerebral artery occlusion.Notably,we observed a shift in tropism from neurons to endothelial cells with post-stroke administration of AAV-PHP.eB-mNeonGreen(mNG).This endothelial cell tropism correlated strongly with expression of the endothelial membrane receptor lymphocyte antigen 6 family member A(Ly6A).Furthermore,AAV-PHP.eB-mediated overexpression of bFGF markedly improved neurobehavioral outcomes and promoted long-term neurogenesis and angiogenesis post-ischemic stroke.Our findings underscore the significance of considering potential tropism shifts when utilizing AAV-PHP.eB-mediated gene therapy in neurological diseases and suggest a promising new strategy for bFGF gene therapy in stroke treatment.展开更多
AIM To investigate the inhibitory effect of astragaloside IV on the pathological functions of cancer-associated fibroblasts,and to explore the underlying mechanism.METHODS Paired gastric normal fibroblast(GNF) and gas...AIM To investigate the inhibitory effect of astragaloside IV on the pathological functions of cancer-associated fibroblasts,and to explore the underlying mechanism.METHODS Paired gastric normal fibroblast(GNF) and gastric cancer-associated fibroblast(GCAF) cultures were established from resected tissues. GCAFs were treated with vehicle control or different concentrations of astragaloside Ⅳ. Conditioned media were prepared from GNFs,GCAFs,control-treated GCAFs,and astragaloside Ⅳ-treated GCAFs,and used to culture BGC-823 human gastric cancer cells. Proliferation,migration and invasion capacities of BGC-823 cells were determined by MTT,wound healing,and Transwell invasion assays,respectively. The action mechanism of astragaloside Ⅳ was investigated by detecting the expression of micro RNAs and the expression and secretion of the oncogenic factor,macrophage colonystimulating factor(M-CSF),and the tumor suppressive factor,tissue inhibitor of metalloproteinase 2(TIMP2),in different groups of GCAFs. The expression of the oncogenic pluripotency factors SOX2 and NANOG in BGC-823 cells cultured with different conditioned media was also examined.RESULTS GCAFs displayed higher capacities to induce BGC-823 cell proliferation,migration,and invasion than GNFs(P < 0.01). Astragaloside Ⅳ treatment strongly inhibited the proliferation-,migration-and invasion-promoting capacities of GCAFs(P < 0.05 for 10 μmol/L,P < 0.01 for 20 μmol/L and 40 μmol/L). Compared with GNFs,GCAFs expressed a lower level of micro RNA-214(P < 0.01) and a higher level of micro RNA-301 a(P < 0.01). Astragaloside Ⅳ treatment significantly upregulated micro RNA-214 expression(P < 0.01) and down-regulated micro RNA-301 a expression(P < 0.01) in GCAFs. Reestablishing the micro RNA expression balance subsequently suppressed M-CSF production(P < 0.01) and secretion(P < 0.05),and elevated TIMP2 production(P < 0.01) and secretion(P < 0.05). Consequently,the ability of GCAFs to increase SOX2 and NANOG expression in BGC-823 cells was abolished by astragaloside Ⅳ.CONCLUSION Astragaloside Ⅳ can inhibit the pathological functions of GCAFs by correcting their dysregulation of micro RNA expression,and it is promisingly a potent therapeutic agent regulating tumor microenvironment.展开更多
Cancer-associated fibroblasts(CAFs) are important components of various types of tumors,including gastric cancer(GC).During tumorigenesis and progression,CAFs play critical roles in tumor invasion and metastasis via a...Cancer-associated fibroblasts(CAFs) are important components of various types of tumors,including gastric cancer(GC).During tumorigenesis and progression,CAFs play critical roles in tumor invasion and metastasis via a series of functions including extracellular matrix deposition,angiogenesis,metabolism reprogramming and chemoresistance.However,the mechanism of the interaction between gastric cancer cells and CAFs remains largely unknown.Micro RNAs(mi RNAs) are a class of non-coding small RNA molecules,and their expression in CAFs not only regulates the expression of a number of target genes but also plays an essential role in the communication between tumor cells and CAFs.In this review,we provide an overview of recent studies on CAF mi RNAs in GC and the relevant signaling pathways in gastrointestinal tumors.Focusing the attention on these signaling pathways may help us better understand their role in tumor invasion and metastasis and identify new molecular targets for therapeutic strategies.展开更多
AIM To study the mechanism of Fuzhenghuayu (FZHY) decoction on anti liver fibrosis. METHODS FZHY 10% decoction sera was incubated with rat normal subcultured hepatic stellate cells (HSC) and fibrotic primarily cul...AIM To study the mechanism of Fuzhenghuayu (FZHY) decoction on anti liver fibrosis. METHODS FZHY 10% decoction sera was incubated with rat normal subcultured hepatic stellate cells (HSC) and fibrotic primarily cultured HSC, normal and fibrotic hepatocytes and subcultured skin fibroblasts separately. Cell intracellular and extracellular collagen synthesis rates were measured by the method of Proline impulse and collagenase digestion. RESULTS For primarily cultured HSC and hepatocytes, both of intracellular and extracellular collagen synthesis rates decreased in the drug sera group. For the normal subcultured HSC and primarily cultured hepatocytes, the extracellular collagen secretion was decreased obviously by the drug sera, and intracellular collagen synthesis rates were inhibited to some extents. For fibroblasts, both intracellular and extracellular collagen synthesis rates were inhibited some what, but no significant differences were found. CONCLUSION The mechanism of FZHY decoction on anti liver fibrosis may be associated with inhibition of liver collagen production.展开更多
To examine the role of gap junctions in cell senescence,the changes of gap junctions in cisplatin-induced premature senescence of primary cultured fibroblasts were studied and compared with the replicative senescent h...To examine the role of gap junctions in cell senescence,the changes of gap junctions in cisplatin-induced premature senescence of primary cultured fibroblasts were studied and compared with the replicative senescent human fibroblasts.Dye transfer assay for gap junction function and immunofluorescent staining for connexin 43 protein distribution were done respectively. Furthermore,cytofluorimetry and DAPI fluorescence staining were performed for cell cycle and apoptosis analysis. p53 gene expression level was detected with indirect immunofluorescence. We found that cisplatin (10 mM) treatment could block cell growth cycle at G1 and induced premature senescence. The premature senescence changes included high frequency of apoptosis,elevation of p53 expression,loss of membranous gap junctions and reduction of dye-transfer capacity. These changes were comparable to the changes of replicative senescence of human fibroblasts. It was also concluded that cisplatin could induce premature senescence concomitant with inhibition of gap junctions in the fibroblasts. Loss of functional gap junctions from the cell membrane may account for the reduced intercellular communication in the premature senescent fibroblasts. The cell system we used may provide a model useful for the study of the gap junction thus promoting agents against premature senescence.展开更多
The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardi...The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardiac fibroblasts(CFs). These CFs were cultured and treated with different concentrations of osthole. Our results showed that the TGF-β1 expression in the CFs transfected with that the recombinant expression plasmids pc DNA3.1(+)-TGF-β1 was significantly enhanced. After the CFs were treated with 1.25-5 μg·m L^(-1) of osthole for 24 h, the m RNA and protein expression levels of collagensⅠand III were reduced. The collagen Ⅰ/III ratio was also reduced. The m RNA and protein expression levels of TGF-β1, TβRⅠ, Smad2/3, P-Smad2/3, Smad4, and α-SMA were decreased, whereas the expression level of Smad7 was increased. These effects suggested that osthole could inhibit collagen Ⅰ and III expression and reduce their ratio via the TGF-β/Smad signaling pathway in TGF-β1 overexpressed CFs. These effects of osthole may play beneficial roles in the prevention and treatment of myocardial fibrosis.展开更多
The hepatic stellate cells in the liver are stimulated sustainably by chronic injury of the hepatocytes, activating myofibroblasts, which produce abundant collagen. Myofibroblasts are the major source of extracellular...The hepatic stellate cells in the liver are stimulated sustainably by chronic injury of the hepatocytes, activating myofibroblasts, which produce abundant collagen. Myofibroblasts are the major source of extracellular proteins during fibrogenesis, and may directly, or secreted products, contribute to carcinogenesis and tumor progression. Cancer-associated fibroblasts(CAFs) are one of the components of the tumor microenvironment that promote the proliferation and invasion of cancer cells by secreting various growth factors and cytokines. CAFs crosstalk with cancer cells stimulates tumor progression by creating a favorable microenvironment for progression, invasion, and metastasis through the epithelial-mesenchymal transition. Basic studies on CAFs have advanced, and the role of CAFs in tumors has been elucidated. In particular, for hepatocellular carcinoma, carcinogenesis from cirrhosis is a known fact, and participation of CAFs in carcinogenesis is supported. In this review, we discuss the current literature on the role of CAFs and CAF-related signaling in carcinogenesis, crosstalk with cancer cells, immunosuppressive effects, angiogenesis, therapeutic targets, and resistance to chemotherapy. The role of CAFs is important in cancer initiation and progression. CAF-targeted therapy may be effective for suppression not only of fibrosis but also cancer progression.展开更多
基金Supported by the National Natural Science Foundation of China(61988102,62401113,92463308)the National Safety Academic Fund(U2130113)+2 种基金the Sichuan Science and Technology Program(2022JDJQ0065)the Chengdu Science and Technology Program(2024-YF05-01803-SN)the Sichuan Provincial Administration of Traditional Chinese Medicine(2024MS512)and the from Key Laboratory of THz Technology,Ministry of Education.
文摘Fibroblasts support a broad range of essential organ functions via microarchitectural,biomechanical,and biochemical cues.Despite great advances in fluorescence,photoacoustic conversion,and Raman scattering over the past decades,their invasiveness and limited spatial resolution hinder the characterization of fibroblasts in a single cell.Here,taking mouse embryonic fibroblasts(MEFs)as an example,we propose a novel noninvasive approach to investigate the compositional distribution of MEFs at the single-cell scale via terahertz(THz)nanos⁃copy.Compared to the topological morphology,THz nano-imaging enables the component-based visualization of MEFs,such as the membrane,cytoplasm,nucleus,and extracellular vesicles(EVs).Notably,we demonstrate the real-space observation of the influence of rapamycin treatment on the increase of EVs in MEFs.Moreover,the line-cut and area-statistical analysis establishes the relationship between the topological morphology and the THz near-field amplitudes for different cellular components of MEFs.This work provides a new pathway to char⁃acterize the effects of pharmaceutical treatments,with potential applications in disease diagnosis and drug devel⁃opment.
基金Supported by National Key Research and Development Program Project,No.2017YFC1700601Shaanxi Provincial Key Research and Development Program Project,No.2018SF-350Leading Talents in Scientific and Technological Innovation of the Shaanxi Province Special Support Plan,No.00518。
文摘Pancreatic ductal adenocarcinoma stands out as an exceptionally fatal cancer owing to the complexities associated with its treatment and diagnosis,leading to a notably low five-year survival rate.This study offers a detailed exploration of epidemiological trends in pancreatic cancer and key molecular drivers,such as mutations in CDKN2A,KRAS,SMAD4,and TP53,along with the influence of cancer-associated fibroblasts(CAFs)on disease progression.In particular,we focused on the pivotal roles of signaling pathways such as the transforming growth factor-βand Wnt/β-catenin pathways in the development of pancreatic cancer and investigated their application in emerging therapeutic strategies.This study provides new scientific perspectives on pancreatic cancer treatment,especially in the development of precision medicine and targeted therapeutic strategies,and demonstrates the importance of signaling pathway research in the development of effective therapeutic regimens.Future studies should explore the subtypes of CAFs and their specific roles in the tumor microenvironment to devise more effective therapeutic methods.
基金supported by the grants provided by Zhuhai People's Hospital,China(Grant No.:2021KYQD-03)the National Natural Science Foundation of China(Grant No.:22176016).
文摘Fibroblast activation protein(FAP)is overexpressed in cancer-associated fibroblasts across various cancer types.Numerous radiolabeled FAP inhibitors(FAPIs)(Fig.S1A)currently under clinical investigation have shown remarkable potential in cancer theranostics.
文摘Breast cancer is one of the most common malignancies worldwide and is a major cause of cancer-related mortality among women.Beyond tumor cells,the tumor microenvironment(TME)also plays an important role in cancer progression,therapy resistance,and metastasis.The TME is a complex ecosystem consisting of stromal and immune cells,extracellular matrix(ECM),and various signaling molecules that dynamically interact with tumor cells.Cancer-associated fibro-blasts remodel the ECM and secrete growth factors that promote tumor growth and invasion.Immune cells,such as tumor-associated macrophages,regulatory T cells,and myeloid-derived suppressor cells,often contribute to an immunosup-pressive environment that hinders anti-tumor immune responses.The ECM pro-vides structural support and acts as a reservoir for signaling molecules that in-fluence cancer cell behavior.These components evolve together with tumor cells,facilitating immune evasion,therapy resistance,and epithelial-to-mesenchymal transition,which promotes metastasis.Understanding these interactions is nece-ssary to develop novel therapeutic strategies that target both tumor and micro-environmental components.This minireview highlights the key stromal and immune elements within the breast cancer microenvironment,discussing their individual and collective roles in tumor progression and clinical outcomes,while emphasizing emerging therapeutic approaches aiming to reprogram the TME to improve treatment efficacy.
基金Supported by National Natural Science Foundation of China,No.82304151.
文摘Pancreatic cancer is a highly aggressive malignancy with a poor prognosis and limited therapeutic options.The tumor microenvironment(TME),including cancer-associated fibroblasts(CAFs),plays a pivotal role in tumor progression and therapy resistance.Senescent CAFs,which exhibit a senescence-associated secretory phenotype(SASP),further exacerbate cancer growth through inflammatory cytokine secretion.This editorial highlights a study by Jiang et al,which investigates the potential of resveratrol,a natural polyphenolic compound,in targeting senescent CAFs to inhibit pancreatic cancer progression.The study demonstrates that resveratrol reduces senescent CAFs and downregulates SASP factors,thereby disrupting the pro-tumorigenic activities of these cells.Resveratrol’s ability to modulate the TME,induce apoptosis in pancreatic cancer cells,and inhibit metastasis underscores its potential as an adjunctive therapy.This research offers promising insights into novel strategies for improving therapeutic outcomes in pancreatic cancer by targeting the TME and senescent CAFs.
文摘The resistance to cancer treatment is a major clinical obstacle,being strongly influenced by the tumor microen-vironment(TME).Cancer-associated fibroblasts(CAFs)are critical elements of the TME.CAFs are heterogeneous and are activated through diverse pathways.These CAFs engage in reciprocal interactions with tumor cells,driv-ing tumor progression and therapeutic resistance.In this review,we discuss the role of CAFs in the development of tumor resistance to chemotherapy,radiotherapy,targeted therapy,and immunotherapy.Besides,we sum-marize recent clinical trials in CAF-targeted therapies.The development of resistance involves physical barrier formation,metabolic reprogramming,exosome release,DNA repair,bypass pathway activation,multidrug resis-tance protein upregulation,and immune checkpoint inhibition.Challenges remain in addressing drug resistance despite the therapeutic potential of targeting CAFs:the cellular origins of CAFs need to be clarified,and their limited clinical applications need to be increased.Future studies should focus on elucidating the reasons for CAF heterogeneity,developing precise targeting strategies,and validating the clinical safety and efficacy of CAF-based therapies to overcome treatment resistance and improve patient outcomes.
文摘Objectives:The present study investigated whether Tripartite Motif-Containing Protein 32(TRIM32)contributes to the aberrant activation of keloid fibroblasts(KFs)via glycolysis.Methods:The expression levels of TRIM32,pyruvate dehydrogenase kinase 1(PDK1),hexokinase 2(HK2),and glucose transporter 1(GLUT1)in normal human skin fibroblasts(NFs)and KFs were analyzed using RT-qPCR analyses and western blotting.Cellular proliferation,invasion,and migration were evaluated using Transwell,wound healing,5-ethynyl-2′-deoxyuridine(EdU),and cell counting kit-8(CCK-8)assays.The extracellular acidification rate(ECAR)was measured using the XF96 Extracellular Flux Analyzer.Glucose uptake and ATP production were measured using specific assay kits.The expression ofα-smooth muscle actin(α-SMA)was determined by immunofluorescence assays.The expression levels of collagen I,α-smooth muscle actin(α-SMA),fibronectin(FN),and components of the phosphoinositide-3-kinase/protein kinase B(PI3K/AKT)signaling pathway were quantified by western blotting.Results:The expression of TRIM32 and glycolysis-related proteins was significantly elevated in KFs compared to that in NFs.TRIM32 overex-pression enhanced the proliferation,invasion,and migration of KFs,as well as extracellular matrix(ECM)deposition,glucose uptake,and ATP production,while TRIM32 silencing produced the opposite effects.The glycolysis inhibitor,2-deoxy-glucose(2-DG),significantly suppressed the biological functions of KFs;however,TRIM32 overexpression effectively counteracted the inhibitory effects of 2-DG.TRIM32 activated the PI3K/AKT signaling pathway in KFs.The PI3K inhibitor LY294002 decreased cellular glycolysis,with TRIM32 overexpression mitigated these inhibitory effects.Conclusion:This study demonstrated that TRIM32 enhances the viability of KFs by regulating glycolytic activity,potentially mediated via the PI3K/AKT signaling pathway,thereby suggesting novel therapeutic approaches for the treatment of keloids.
基金supported by the National Natural Science Foundation of China(82373037 and 82403486)the Natural Science Foundation of Jiangsu Province(BK20230054 and BK20230161)+1 种基金the China Postdoctoral Science Foundation(2023M741766)the Nanjing Medical Science and Technology Development Foundation,Nanjing Department of Health(YKK21182 and JQX23010).
文摘Perineural invasion(PNI)by tumor cells is a key phenotype of highly-invasive oral squamous cell carcinoma(OSCC).Since Schwann cells(SCs)and fibroblasts maintain the physiological homeostasis of the peripheral nervous system,and we have focused on cancer-associated fibroblasts(CAFs)for decades,it’s imperative to elucidate the impact of CAFs on SCs in PNI+OSCCs.We describe a disease progression-driven shift of PNI−towards PNI+during the progression of early-stage OSCC(31%,n=125)to late-stage OSCC(53%,n=97),characterized by abundant CAFs and nerve demyelination.CAFs inhibited SC proliferation/migration and reduced neurotrophic factors and myelin in vitro,and this involved up-regulated ER stress and decreased MAPK signals.Moreover,CAFs also aggravated the paralysis of the hind limb and PNI in vivo.Unexpectedly,leukemia inhibitory factor(LIF)was exclusively expressed on CAFs and up-regulated in metastatic OSCC.The LIF inhibitor EC330 restored CAF-induced SC inactivation.Thus,OSCC-derived CAFs inactivate SCs to aggravate nerve injury and PNI development.
基金supported by 1.3.5 project for disciplines of excellence,West China Hospital,Sichuan University(ZYJC21048)Foundation of Sichuan Provincial Science and Technology Program(2022JDR0091,2023NSFSC0004,2023NSFSC0639,2023NSFSC1742)+5 种基金Cooperation Project for Academician&Expert Workstation(HXYS20001)Sichuan University Education Foundation(0040206107011)National Natural Science Foundation of China(Nos.82371883,82402191)China Postdoctoral Science Foundation(2023M732456)Postdoctor Research Fund of West China Hospital(2024HXBH142)Sichuan University“From 0 to 1”Innovation Research Project(2023SCUH0056).
文摘Objectives:Recently,pre-/post-operative Local Estrogen Therapy(LET)has shown effectiveness in alleviating Pelvic Organ Prolapse(POP)symptoms in clinical therapy.However,there is a lack of scientific evidence to support these claims.Therefore,we aimed to explore the anti-senescence effects and mechanisms of 17β-estradiol(E2)on POP-derived fibroblasts.Methods:The primary fibroblast cells were isolated and cultured fromthe surgical samples of postmenopausal women clinically diagnosed with pelvic organ prolapse(POP)at stages III-IV(quantified using the POP-Q system)and without any other treatment within 6 months.(n=12,age 50–75).Colorimetric Cell Counting Kit(CCK-8)assay and Senescence-Associated-β-Galactosidase(SA-β-Gal)staining were used to test the cell proliferative capacity and the senescence rate.Western blotting(WB)was used to detect the expression of Collagen Type I(COL-I),Collagen Type III(COL-III),Cyclin-dependent kinase 4 inhibitor A(p16INK4a),Cyclin-dependent kinase inhibitor 1A(p21),Tumor Protein 53(p53),Sirtuin 1(SIRT-1)and Microtubule-associated protein 1A/1B-light chain 3-I/II(LC3-I/II)protein.A transmission ElectronMicroscope(TEM)was used to observe the ultrastructure of fibroblasts.Results:The results showed that E2 significantly promoted the proliferation of fibroblasts derived from POP and reduced the staining rate of SA-β-Gal.It markedly enhanced the extracellular matrix proteins COL-I and COL-III,accompanied by inhibition of the senescent maker p16INK4a.Additionally,our results improved the cells’autophagy and metabolic activity.Additionally,our results indicate the anti-senescence mechanism of E2 through the mediated SIRT-1/p53/p21 axis pathway.Conclusion:We provide preliminary evidence for the anti-aging effects and mechanisms of E2 on POP,hoping to provide a theoretical basis for estrogen against POP senescence and guide the clinical application and local administration of estrogen in POP treatment.
基金supported by the National Natural Science Foundation of China(grant number 81773285)Beijing Chao-Yang Hospital Golden Seeds Foundation(grant number CYJZ202341).
文摘Osteosarcoma(OS)is a prevalent primary bone malignancy with limited treatment options.Therefore,it is imperative to investigate and understand the mechanisms underlying OS pathogenesis.Cancer-associated fibroblasts(CAFs)are markedly abundant in tumor stromal cells and are essentially involved in the modulation of tumor occurrence and development.In recent years,CAFs have become a hotspot as researchers aim to elucidate CAF mechanisms that regulate tumor progression.However,most studies on CAFs are limited to a few common cancers,and their association with OS remains elusive.This review describes the role and current knowledge of CAFs in OS,focusing on their potential cellular origin,classification,and diverse functionality.It was found that CAFs influenced OS tumor cell signaling,proliferation,invasion,metastasis,epithelial-mesenchymal transition,stemness maintenance,angiogenesis,and the ability to modify immune system components.Furthermore,findings on other common cancers indicated that effective therapeutic strategies included the manipulation of CAF activation,targeting CAF-derived components,and depletion of CAFs by biomarkers.This review provides new insights and a theoretical basis for OS research.
基金Supported by the National Natural Science Foundation of China(No.81770898).
文摘AIM:To explore the effect of co-host non-coding RNA(ncRNA)MIR503HG/miR-503-5p on the angiogenesis of pterygium.METHODS:MIR503HG/miR-503-5p/fibroblast growth factor 2(FGF2)expression levels in pterygium tissues,control conjunctival tissues,and human pterygium fibroblasts(HPF)were examined by reverse transcription-polymerase chain reaction(qRT-PCR)and immunohistochemical methods.Effects of MIR503HG/miR-503-5p on low molecular weight FGF2(LWM FGF2),migration and angiogenesis of human retinal microvascular endothelial cells(HRMEC)were determined in an HPF and HRMEC co-culture model using Western blots,wound healing assay,Matrigel-based tube formation assay,and Transwell assay.RESULTS:MIR503HG/miR-503-5p/FGF2 pathway was actively increased in pterygium tissue and there was a negative correlation between the expression of the two ncRNAs.FGF2 expression level was positively correlated with MIR503HG and negatively correlated with miR-503-5p.Overexpressed MIR503HG/miR-503-5p did not affect the migration and angiogenesis of HRMECs cultured separately,but significantly affected migration and angiogenesis of HRMEC in HPF and HRMEC co-culture models.Western blotting revealed that MIR503HG/miR-503-5p overexpression significantly increased LMW FGF2 expression in HPF.CONCLUSION:MIR503HG/miR-503-5p inhibits HRMEC migration and angiogenic function by interfering with the interaction between HPF and endothelial cells via reducing LMW FGF2 in HPF.
基金Supported by National Natural Science Foundation of China(30801353)Shandong Education Department Foundation Project(G08LG53)~~
文摘[Objective] The aim of this study was to establish the in vitro culture system of chicken fibroblasts.[Method] Tissue explant method and enzymatic digestion method were used to separate and culture chicken skin fibroblasts respectively.The rate of cell growth,cryopreservation and recovery were compared.[Result] The primary chicken fibroblasts prepared by enzymatic digestion grew faster and converged together to form monolayer on 5 d post preparation;the passage cells prepared by these 2 methods grew at similar speed and formed monolayer within 2-3 d;homogeneous fibroblasts could be obtained by trypsin digestion and repeated attachment for 3-4 passages;there were 75%-80% of cells survived after cryopreservation and recovery;the growth curves of embryonic fibroblasts and skin fibroblasts were all normal and the two kind of cells still retained the normal number of chromosomes even at the twelfth passage.[Conclusion] The feeder layer cells needed for establishing ES cell lines could be obtained by culturing chicken fibroblasts through both tissue explant method and enzymatic digestion method.This study provided a basis for the successful establishment of ES cell lines.
基金supported by the National Natural Science Foundation of China,Nos.82171456(to QY)and 81971229(to QY)the Natural Science Foundation of Chongqing,Nos.CSTC2021JCYJ-MSXMX0263(to QY)and CSTB2023NSCQ-MSX1015(to XL)Doctoral Innovation Project of The First Affiliated Hospital of Chongqing Medical University,Nos.CYYY-BSYJSCXXM-202318(to JW)and CYYY-BSYJSCXXM-202327(to HT).
文摘Recent studies have shown that fibrotic scar formation following cerebral ischemic injury has varying effects depending on the microenvironment.However,little is known about how fibrosis is induced and regulated after cerebral ischemic injury.Sonic hedgehog signaling participates in fibrosis in the heart,liver,lung,and kidney.Whether Shh signaling modulates fibrotic scar formation after cerebral ischemic stroke and the underlying mechanisms are unclear.In this study,we found that Sonic Hedgehog expression was upregulated in patients with acute ischemic stroke and in a middle cerebral artery occlusion/reperfusion injury rat model.Both Sonic hedgehog and Mitofusin 2 showed increased expression in the middle cerebral artery occlusion rat model and in vitro fibrosis cell model induced by transforming growth factor-beta 1.Activation of the Sonic hedgehog signaling pathway enhanced the expression of phosphorylated Smad 3 and Mitofusin 2 proteins,promoted the formation of fibrotic scars,protected synapses or promoted synaptogenesis,alleviated neurological deficits following middle cerebral artery occlusion/reperfusion injury,reduced cell apoptosis,facilitated the transformation of meninges fibroblasts into myofibroblasts,and enhanced the proliferation and migration of meninges fibroblasts.The Smad3 phosphorylation inhibitor SIS3 reversed the effects induced by Sonic hedgehog signaling pathway activation.Bioinformatics analysis revealed significant correlations between Sonic hedgehog and Smad3,between Sonic hedgehog and Mitofusin 2,and between Smad3 and Mitofusin 2.These findings suggest that Sonic hedgehog signaling may influence Mitofusin 2 expression by regulating Smad3 phosphorylation,thereby modulating the formation of early fibrotic scars following cerebral ischemic stroke and affecting prognosis.The Sonic Hedgehog signaling pathway may serve as a new therapeutic target for stroke treatment.
基金supported by the National Natural Science Foundation of China,Nos.81870921(to YW),81974179(to ZZ),82271320(to ZZ),82071284(to YT)National Key R&D Program of China,No.2022YFA1603600(to ZZ),2019YFA0112000(to YT)+1 种基金Scientific Research and Innovation Program of Shanghai Education Commission,No.2019-01-07-00-02-E00064(to GYY)Scientific and Technological Innovation Act Program of Shanghai Science and Technology Commission,No.20JC1411900(to GYY).
文摘AAV-PHP.eB is an artificial adeno-associated virus(AAV)that crosses the blood-brain barrier and targets neurons more efficiently than other AAVs when administered systematically.While AAV-PHP.eB has been used in various disease models,its cellular tropism in cerebrovascular diseases remains unclear.In the present study,we aimed to elucidate the tropism of AAV-PHP.eB for different cell types in the brain in a mouse model of ischemic stroke and evaluate its effectiveness in mediating basic fibroblast growth factor(bFGF)gene therapy.Mice were injected intravenously with AAV-PHP.eB either 14 days prior to(pre-stroke)or 1 day following(post-stroke)transient middle cerebral artery occlusion.Notably,we observed a shift in tropism from neurons to endothelial cells with post-stroke administration of AAV-PHP.eB-mNeonGreen(mNG).This endothelial cell tropism correlated strongly with expression of the endothelial membrane receptor lymphocyte antigen 6 family member A(Ly6A).Furthermore,AAV-PHP.eB-mediated overexpression of bFGF markedly improved neurobehavioral outcomes and promoted long-term neurogenesis and angiogenesis post-ischemic stroke.Our findings underscore the significance of considering potential tropism shifts when utilizing AAV-PHP.eB-mediated gene therapy in neurological diseases and suggest a promising new strategy for bFGF gene therapy in stroke treatment.
基金Supported by the National Natural Science Foundation of China,No.81760552the Program of the Inner Mongolia Natural Science Foundation,No.2016MS0824 and No.2015MS0896+1 种基金the Program of“Keji Baiwan Gongcheng”of Inner Mongolia Medical University,No.YKD2015KJBW008the Supporting Program for Outstanding Youth in Science and Technology of Inner Mongolia Autonomous Region,No.NJYT-17-B30
文摘AIM To investigate the inhibitory effect of astragaloside IV on the pathological functions of cancer-associated fibroblasts,and to explore the underlying mechanism.METHODS Paired gastric normal fibroblast(GNF) and gastric cancer-associated fibroblast(GCAF) cultures were established from resected tissues. GCAFs were treated with vehicle control or different concentrations of astragaloside Ⅳ. Conditioned media were prepared from GNFs,GCAFs,control-treated GCAFs,and astragaloside Ⅳ-treated GCAFs,and used to culture BGC-823 human gastric cancer cells. Proliferation,migration and invasion capacities of BGC-823 cells were determined by MTT,wound healing,and Transwell invasion assays,respectively. The action mechanism of astragaloside Ⅳ was investigated by detecting the expression of micro RNAs and the expression and secretion of the oncogenic factor,macrophage colonystimulating factor(M-CSF),and the tumor suppressive factor,tissue inhibitor of metalloproteinase 2(TIMP2),in different groups of GCAFs. The expression of the oncogenic pluripotency factors SOX2 and NANOG in BGC-823 cells cultured with different conditioned media was also examined.RESULTS GCAFs displayed higher capacities to induce BGC-823 cell proliferation,migration,and invasion than GNFs(P < 0.01). Astragaloside Ⅳ treatment strongly inhibited the proliferation-,migration-and invasion-promoting capacities of GCAFs(P < 0.05 for 10 μmol/L,P < 0.01 for 20 μmol/L and 40 μmol/L). Compared with GNFs,GCAFs expressed a lower level of micro RNA-214(P < 0.01) and a higher level of micro RNA-301 a(P < 0.01). Astragaloside Ⅳ treatment significantly upregulated micro RNA-214 expression(P < 0.01) and down-regulated micro RNA-301 a expression(P < 0.01) in GCAFs. Reestablishing the micro RNA expression balance subsequently suppressed M-CSF production(P < 0.01) and secretion(P < 0.05),and elevated TIMP2 production(P < 0.01) and secretion(P < 0.05). Consequently,the ability of GCAFs to increase SOX2 and NANOG expression in BGC-823 cells was abolished by astragaloside Ⅳ.CONCLUSION Astragaloside Ⅳ can inhibit the pathological functions of GCAFs by correcting their dysregulation of micro RNA expression,and it is promisingly a potent therapeutic agent regulating tumor microenvironment.
文摘Cancer-associated fibroblasts(CAFs) are important components of various types of tumors,including gastric cancer(GC).During tumorigenesis and progression,CAFs play critical roles in tumor invasion and metastasis via a series of functions including extracellular matrix deposition,angiogenesis,metabolism reprogramming and chemoresistance.However,the mechanism of the interaction between gastric cancer cells and CAFs remains largely unknown.Micro RNAs(mi RNAs) are a class of non-coding small RNA molecules,and their expression in CAFs not only regulates the expression of a number of target genes but also plays an essential role in the communication between tumor cells and CAFs.In this review,we provide an overview of recent studies on CAF mi RNAs in GC and the relevant signaling pathways in gastrointestinal tumors.Focusing the attention on these signaling pathways may help us better understand their role in tumor invasion and metastasis and identify new molecular targets for therapeutic strategies.
文摘AIM To study the mechanism of Fuzhenghuayu (FZHY) decoction on anti liver fibrosis. METHODS FZHY 10% decoction sera was incubated with rat normal subcultured hepatic stellate cells (HSC) and fibrotic primarily cultured HSC, normal and fibrotic hepatocytes and subcultured skin fibroblasts separately. Cell intracellular and extracellular collagen synthesis rates were measured by the method of Proline impulse and collagenase digestion. RESULTS For primarily cultured HSC and hepatocytes, both of intracellular and extracellular collagen synthesis rates decreased in the drug sera group. For the normal subcultured HSC and primarily cultured hepatocytes, the extracellular collagen secretion was decreased obviously by the drug sera, and intracellular collagen synthesis rates were inhibited to some extents. For fibroblasts, both intracellular and extracellular collagen synthesis rates were inhibited some what, but no significant differences were found. CONCLUSION The mechanism of FZHY decoction on anti liver fibrosis may be associated with inhibition of liver collagen production.
基金supported by grants from The National Key Program for Basic Research,Project No.G2000057002National Natural Science Foudation of China,Project No.30270685
文摘To examine the role of gap junctions in cell senescence,the changes of gap junctions in cisplatin-induced premature senescence of primary cultured fibroblasts were studied and compared with the replicative senescent human fibroblasts.Dye transfer assay for gap junction function and immunofluorescent staining for connexin 43 protein distribution were done respectively. Furthermore,cytofluorimetry and DAPI fluorescence staining were performed for cell cycle and apoptosis analysis. p53 gene expression level was detected with indirect immunofluorescence. We found that cisplatin (10 mM) treatment could block cell growth cycle at G1 and induced premature senescence. The premature senescence changes included high frequency of apoptosis,elevation of p53 expression,loss of membranous gap junctions and reduction of dye-transfer capacity. These changes were comparable to the changes of replicative senescence of human fibroblasts. It was also concluded that cisplatin could induce premature senescence concomitant with inhibition of gap junctions in the fibroblasts. Loss of functional gap junctions from the cell membrane may account for the reduced intercellular communication in the premature senescent fibroblasts. The cell system we used may provide a model useful for the study of the gap junction thus promoting agents against premature senescence.
基金supported by the National Nature Science Foundation of China(No.81302772)Jiangsu Provincial Medical Talent Project(No.QNRC2016716)
文摘The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardiac fibroblasts(CFs). These CFs were cultured and treated with different concentrations of osthole. Our results showed that the TGF-β1 expression in the CFs transfected with that the recombinant expression plasmids pc DNA3.1(+)-TGF-β1 was significantly enhanced. After the CFs were treated with 1.25-5 μg·m L^(-1) of osthole for 24 h, the m RNA and protein expression levels of collagensⅠand III were reduced. The collagen Ⅰ/III ratio was also reduced. The m RNA and protein expression levels of TGF-β1, TβRⅠ, Smad2/3, P-Smad2/3, Smad4, and α-SMA were decreased, whereas the expression level of Smad7 was increased. These effects suggested that osthole could inhibit collagen Ⅰ and III expression and reduce their ratio via the TGF-β/Smad signaling pathway in TGF-β1 overexpressed CFs. These effects of osthole may play beneficial roles in the prevention and treatment of myocardial fibrosis.
基金Supported by Research Program on Hepatitis from Japan Agency for Medical Research and development,AMED
文摘The hepatic stellate cells in the liver are stimulated sustainably by chronic injury of the hepatocytes, activating myofibroblasts, which produce abundant collagen. Myofibroblasts are the major source of extracellular proteins during fibrogenesis, and may directly, or secreted products, contribute to carcinogenesis and tumor progression. Cancer-associated fibroblasts(CAFs) are one of the components of the tumor microenvironment that promote the proliferation and invasion of cancer cells by secreting various growth factors and cytokines. CAFs crosstalk with cancer cells stimulates tumor progression by creating a favorable microenvironment for progression, invasion, and metastasis through the epithelial-mesenchymal transition. Basic studies on CAFs have advanced, and the role of CAFs in tumors has been elucidated. In particular, for hepatocellular carcinoma, carcinogenesis from cirrhosis is a known fact, and participation of CAFs in carcinogenesis is supported. In this review, we discuss the current literature on the role of CAFs and CAF-related signaling in carcinogenesis, crosstalk with cancer cells, immunosuppressive effects, angiogenesis, therapeutic targets, and resistance to chemotherapy. The role of CAFs is important in cancer initiation and progression. CAF-targeted therapy may be effective for suppression not only of fibrosis but also cancer progression.
