Objective Long non-coding RNAs(lncRNAs)are critical in the pathogenesis of hematological malignancies,including acute myeloid leukemia(AML).However,the specific role and underlying mechanisms of the lncRNA chromosome ...Objective Long non-coding RNAs(lncRNAs)are critical in the pathogenesis of hematological malignancies,including acute myeloid leukemia(AML).However,the specific role and underlying mechanisms of the lncRNA chromosome 9 open reading frame 139(C9orf139)in AML remain unclear.This study aimed to investigate the role and molecular mechanism of C9orf139 in AML development.Methods AML-related sequencing and microarray data were retrieved from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases.Significant lncRNAs and mRNAs influencing AML progression were identified and analyzed.A competing endogenous RNA network involving lncRNA–microRNA(miRNA)3–mRNA interactions was subsequently constructed.The expression levels of C9orf139,miR-24-3p,and human TAO kinase 1(TAOK1)were assessed via real-time fluorescent quantitative polymerase chain reaction(PCR).Cell proliferation was evaluated via the Cell Counting Kit-8(CCK8)assay,whereas Transwell assays were used to assess cell invasion and migration.Apoptosis was measured by Annexin V Fluorescein Isothiocyanate(FITC)double staining.Tumor formation in nude mice was assessed to examine the effect of C9orf139 on in vivo tumor growth.The C9orf139-miR-24-3p-TAOK1 regulatory axis was validated via dual luciferase reporter assays and RNA-binding protein immunoprecipitation(RIP).Western blot assays were used to assess the expression and phosphorylation of key proteins in the mitogen-activated protein kinase(MAPK)signaling pathway.Results Bioinformatics analysis identified C9orf139 and TAOK1 as differentially expressed genes that play key roles in AML pathogenesis.The C9orf139-miR-24-3p-TAOK1 axis was tightly linked to AML development,as confirmed by clinical sample analysis.In vitro,C9orf139 downregulation resulted in reduced proliferation,invasion,and migration and enhanced the apoptosis of AML cells.In vivo,the inhibition of C9orf139 significantly impaired tumor growth in nude mice.The regulatory axis was further validated.C9orf139 knockdown reduced the phosphorylation levels of the key MAPK pathway proteins,including Raf,mitogen-activated protein kinase kinase(MEK),and extracellular regulated protein kinase(ERK).Conclusion C9orf139 regulates AML progression by activating the MAPK signaling pathway through the C9orf139-miR-24-3p-TAOK1 axis.展开更多
Mesenchymal stem cell （MSC）-based treatments have shown promise for improving tendon healing and repair. MSCs have the potential to differentiate into multiple lineages in response to select chemical and physical st...Mesenchymal stem cell （MSC）-based treatments have shown promise for improving tendon healing and repair. MSCs have the potential to differentiate into multiple lineages in response to select chemical and physical stimuli, including into tenocytes. Cell elongation and cytoskeletal tension have been shown to be instrumental to the process of MSC differentiation. Previous studies have shown that inhibition of stress fiber formation leads MSCs to default toward an adipogenic lineage, which suggests that stress fibers are required for MSCs to sense the environmental factors that can induce differentiation into tenocytes. As the Rho/ROCK signal transduction pathway plays a critical role in both stress fiber formation and in cell sensation, we examined whether the activation of this pathway was required when inducing MSC tendon differentiation using rope-like silk scaffolds. To accomplish this, we employed a loss-of-function approach by knocking out ROCK, actin and myosin （two other components of the pathway） using the specific inhibitors Y-27632, Latrunculin A and blebbistatin, respectively. We demonstrated that independently disrupting the cytoskeleton and the Rho/ ROCK pathway abolished the expression of tendon differentiation markers and led to a loss of spindle morphology. Together, these studies suggest that the tension that is generated by MSC elongation is essential for MSC teno-differentiation and that the Rho/ROCK pathway is a critical mediator of tendon differentiation on rope-like silk scaffolds.展开更多
Pancreatic Ductal Adenocarcinoma(PDAC)is one of the most lethal human malignancies with poor survival outcome.It is characterized by late diagnosis,rapid growth and limited therapeutic choices.Increasing evidence has ...Pancreatic Ductal Adenocarcinoma(PDAC)is one of the most lethal human malignancies with poor survival outcome.It is characterized by late diagnosis,rapid growth and limited therapeutic choices.Increasing evidence has suggested that gene expression may undergo extensive changes during the process of normal cells to malignant transformation which may play a critical role in the initiation and progression of human PDAC.In this study,we applied single-cell RNA sequencing(scRNA-seq)technology to comprehensively compared the Peripheral Blood Mononuclear Cells(PBMCs)and tissue samples from pancreatic ductal adenocarcinoma patients and normal individuals.We characterized transcriptional heterogeneity and identified differentially expressed genes as well as the changes of immune microenvironment between PDAC and normal individuals through integrative bioinformatic analysis.We believe that our results will provide new insights into the cellular and molecular characterization of PDAC and may provide potential implications for future therapeutic approaches and biomarker discoveries.展开更多
Protein interacting with C kinase 1(PICK1)interacts with a variety of membrane proteins and receptors involved in nervous system diseases and multiple cancers.However,the role of PICK1 in gastric cancer remains unclea...Protein interacting with C kinase 1(PICK1)interacts with a variety of membrane proteins and receptors involved in nervous system diseases and multiple cancers.However,the role of PICK1 in gastric cancer remains unclear.In the present work,we explored the expression and interactions of PICK 1 with Toll-like receptor 4(TLR4)in gastric cancer.Clinical data analysis showed that PICK1 expression decreases and is predictive of worse outcomes in patients with gastric cancer.High PICK1 levels attenuate the proliferation and migration of gastric cancer cells,which is dependent on the TLR4/myeloid differentiation primary response 88(MyD88)signaling pathway.Furthermore,in vitro experiments demonstrated that PICK1 affects the trafficking and degradation of TLR4 and promotes TLR4 degradation via autophagy in gastric cancer cells.Molecular dynamics simulations highlighted the binding strength and stability of the TLR4-PICK1 complex.Our study provides new insights into the cellular and pathological functions of PICK1 in gastric cancer.展开更多
Objective:To investigate whether electroacupuncture(EA)alleviates cognitive impairment by suppressing the toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)signaling pathway,which triggers immune-infl...Objective:To investigate whether electroacupuncture(EA)alleviates cognitive impairment by suppressing the toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)signaling pathway,which triggers immune-inflammatory responses in the hippocampus of rats with vascular dementia(VaD).Methods:The experiments were conducted in 3 parts and in total the Sprague-Dawley rats were randomly divided into 8 groups by a random number table,including sham,four-vessel occlusion(4-VO),4-VO+EA,4-VO+non-EA,sham+EA,4-VO+lipopolysaccharide(LPS),4-VO+LPS+EA,and 4-VO+TAK-242 groups.The VaD model was established by the 4-VO method.Seven days later,rats were treated with EA at 5 acupoints of Baihui(DV 20),Danzhong(RN 17),Geshu(BL 17),Qihai(RN 6)and Sanyinjiao(SP 6),once per day for 3 consecutive weeks.Lymphocyte subsets,lymphocyte transformation rates,and inflammatory cytokines interleukin-6(IL-6)and tumor necrosis factorα(TNF-α)were measured to assess immune function and inflammation in VaD rats.Transmission electron microscopy was used to observe the ultrastructure of nerve cells in the hippocampus.The levels of TLR4,MyD88,IL-6,and TNF-αwere detected after EA treatment.TLR4/MyD88 signaling and cognitive function were also assessed after intracerebroventricular injection of TLR4 antagonist TAK-242 or TLR4 agonist LPS with or without EA.Results:Compared with the 4-VO group,EA notably improved immune function of rats in the 4-VO+EA group,inhibited the protein and mRNA expressions of TLR4 and MyD88 in the hippocampus of rats,reduced the expressions of serum IL-6 and TNF-α(all P<0.05 or P<0.01),and led to neuronal repair in the hippocampus.There were no significant differences between the 4-VO+LPS+EA and 4-VO+EA groups,nor between the 4-VO+TAK-242 and 4-VO+EA groups(P>0.05).Conclusions:EA attenuated cognitive impairment associated with immune inflammation by inhibition of the TLR4/MyD88 signaling pathway.Thus,EA may be a promising alternative therapy for the treatment of VaD.展开更多
基金supported by the Youth Talent Science and Technology Project of Health Commission of Changzhou(QN202223)the Education Research Project of Nanjing Medical University(2023ZC086)the Changzhou Medical Center(CMCB202422).
文摘Objective Long non-coding RNAs(lncRNAs)are critical in the pathogenesis of hematological malignancies,including acute myeloid leukemia(AML).However,the specific role and underlying mechanisms of the lncRNA chromosome 9 open reading frame 139(C9orf139)in AML remain unclear.This study aimed to investigate the role and molecular mechanism of C9orf139 in AML development.Methods AML-related sequencing and microarray data were retrieved from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases.Significant lncRNAs and mRNAs influencing AML progression were identified and analyzed.A competing endogenous RNA network involving lncRNA–microRNA(miRNA)3–mRNA interactions was subsequently constructed.The expression levels of C9orf139,miR-24-3p,and human TAO kinase 1(TAOK1)were assessed via real-time fluorescent quantitative polymerase chain reaction(PCR).Cell proliferation was evaluated via the Cell Counting Kit-8(CCK8)assay,whereas Transwell assays were used to assess cell invasion and migration.Apoptosis was measured by Annexin V Fluorescein Isothiocyanate(FITC)double staining.Tumor formation in nude mice was assessed to examine the effect of C9orf139 on in vivo tumor growth.The C9orf139-miR-24-3p-TAOK1 regulatory axis was validated via dual luciferase reporter assays and RNA-binding protein immunoprecipitation(RIP).Western blot assays were used to assess the expression and phosphorylation of key proteins in the mitogen-activated protein kinase(MAPK)signaling pathway.Results Bioinformatics analysis identified C9orf139 and TAOK1 as differentially expressed genes that play key roles in AML pathogenesis.The C9orf139-miR-24-3p-TAOK1 axis was tightly linked to AML development,as confirmed by clinical sample analysis.In vitro,C9orf139 downregulation resulted in reduced proliferation,invasion,and migration and enhanced the apoptosis of AML cells.In vivo,the inhibition of C9orf139 significantly impaired tumor growth in nude mice.The regulatory axis was further validated.C9orf139 knockdown reduced the phosphorylation levels of the key MAPK pathway proteins,including Raf,mitogen-activated protein kinase kinase(MEK),and extracellular regulated protein kinase(ERK).Conclusion C9orf139 regulates AML progression by activating the MAPK signaling pathway through the C9orf139-miR-24-3p-TAOK1 axis.
文摘Mesenchymal stem cell （MSC）-based treatments have shown promise for improving tendon healing and repair. MSCs have the potential to differentiate into multiple lineages in response to select chemical and physical stimuli, including into tenocytes. Cell elongation and cytoskeletal tension have been shown to be instrumental to the process of MSC differentiation. Previous studies have shown that inhibition of stress fiber formation leads MSCs to default toward an adipogenic lineage, which suggests that stress fibers are required for MSCs to sense the environmental factors that can induce differentiation into tenocytes. As the Rho/ROCK signal transduction pathway plays a critical role in both stress fiber formation and in cell sensation, we examined whether the activation of this pathway was required when inducing MSC tendon differentiation using rope-like silk scaffolds. To accomplish this, we employed a loss-of-function approach by knocking out ROCK, actin and myosin （two other components of the pathway） using the specific inhibitors Y-27632, Latrunculin A and blebbistatin, respectively. We demonstrated that independently disrupting the cytoskeleton and the Rho/ ROCK pathway abolished the expression of tendon differentiation markers and led to a loss of spindle morphology. Together, these studies suggest that the tension that is generated by MSC elongation is essential for MSC teno-differentiation and that the Rho/ROCK pathway is a critical mediator of tendon differentiation on rope-like silk scaffolds.
文摘Pancreatic Ductal Adenocarcinoma(PDAC)is one of the most lethal human malignancies with poor survival outcome.It is characterized by late diagnosis,rapid growth and limited therapeutic choices.Increasing evidence has suggested that gene expression may undergo extensive changes during the process of normal cells to malignant transformation which may play a critical role in the initiation and progression of human PDAC.In this study,we applied single-cell RNA sequencing(scRNA-seq)technology to comprehensively compared the Peripheral Blood Mononuclear Cells(PBMCs)and tissue samples from pancreatic ductal adenocarcinoma patients and normal individuals.We characterized transcriptional heterogeneity and identified differentially expressed genes as well as the changes of immune microenvironment between PDAC and normal individuals through integrative bioinformatic analysis.We believe that our results will provide new insights into the cellular and molecular characterization of PDAC and may provide potential implications for future therapeutic approaches and biomarker discoveries.
基金supported by the National Natural Science Foundation of China(No.82172363)the Traditional Chinese Medicine Scientific Research Project of Zhejiang Province(No.2023ZL010)the Zhejiang Medical and Health Science and Technology Project(Nos.2022KY529 and 2024KY636),China.
文摘Protein interacting with C kinase 1(PICK1)interacts with a variety of membrane proteins and receptors involved in nervous system diseases and multiple cancers.However,the role of PICK1 in gastric cancer remains unclear.In the present work,we explored the expression and interactions of PICK 1 with Toll-like receptor 4(TLR4)in gastric cancer.Clinical data analysis showed that PICK1 expression decreases and is predictive of worse outcomes in patients with gastric cancer.High PICK1 levels attenuate the proliferation and migration of gastric cancer cells,which is dependent on the TLR4/myeloid differentiation primary response 88(MyD88)signaling pathway.Furthermore,in vitro experiments demonstrated that PICK1 affects the trafficking and degradation of TLR4 and promotes TLR4 degradation via autophagy in gastric cancer cells.Molecular dynamics simulations highlighted the binding strength and stability of the TLR4-PICK1 complex.Our study provides new insights into the cellular and pathological functions of PICK1 in gastric cancer.
基金the National Natural Science Foundation of China(No.81960811)the Major Research Project of Innovation Group of Guizhou Provincial Department of Education(No.2018KY023)。
文摘Objective:To investigate whether electroacupuncture(EA)alleviates cognitive impairment by suppressing the toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)signaling pathway,which triggers immune-inflammatory responses in the hippocampus of rats with vascular dementia(VaD).Methods:The experiments were conducted in 3 parts and in total the Sprague-Dawley rats were randomly divided into 8 groups by a random number table,including sham,four-vessel occlusion(4-VO),4-VO+EA,4-VO+non-EA,sham+EA,4-VO+lipopolysaccharide(LPS),4-VO+LPS+EA,and 4-VO+TAK-242 groups.The VaD model was established by the 4-VO method.Seven days later,rats were treated with EA at 5 acupoints of Baihui(DV 20),Danzhong(RN 17),Geshu(BL 17),Qihai(RN 6)and Sanyinjiao(SP 6),once per day for 3 consecutive weeks.Lymphocyte subsets,lymphocyte transformation rates,and inflammatory cytokines interleukin-6(IL-6)and tumor necrosis factorα(TNF-α)were measured to assess immune function and inflammation in VaD rats.Transmission electron microscopy was used to observe the ultrastructure of nerve cells in the hippocampus.The levels of TLR4,MyD88,IL-6,and TNF-αwere detected after EA treatment.TLR4/MyD88 signaling and cognitive function were also assessed after intracerebroventricular injection of TLR4 antagonist TAK-242 or TLR4 agonist LPS with or without EA.Results:Compared with the 4-VO group,EA notably improved immune function of rats in the 4-VO+EA group,inhibited the protein and mRNA expressions of TLR4 and MyD88 in the hippocampus of rats,reduced the expressions of serum IL-6 and TNF-α(all P<0.05 or P<0.01),and led to neuronal repair in the hippocampus.There were no significant differences between the 4-VO+LPS+EA and 4-VO+EA groups,nor between the 4-VO+TAK-242 and 4-VO+EA groups(P>0.05).Conclusions:EA attenuated cognitive impairment associated with immune inflammation by inhibition of the TLR4/MyD88 signaling pathway.Thus,EA may be a promising alternative therapy for the treatment of VaD.
