Objectives:Glioblastoma is a prevalent malignant brain tumor,and the actions of the long non-coding RNA HOXA10-AS in its invasion and migration remain unclear.Here,the function of HOXA10-AS in glioblastoma cell invasi...Objectives:Glioblastoma is a prevalent malignant brain tumor,and the actions of the long non-coding RNA HOXA10-AS in its invasion and migration remain unclear.Here,the function of HOXA10-AS in glioblastoma cell invasion and migration and associated mechanisms were investigated.Methods:HOXA10-AS was knocked down in glioblastoma cells,and Transwell and wound healing assays were conducted to elucidate its impacts on cell invasion and migration.Western blotting and quantitative reverse transcription polymerase chain reaction(qRTPCR)assessed HOXA10-AS’s impact on the epithelial-mesenchymal transition(EMT).Microarray analysis identified differentially expressed genes,complemented by bioinformatics approaches to explore potentialmolecular participants and pathways.Rescue experiments validated our findings.Results:HOXA10-AS knockdown significantly inhibits glioblastoma cell migration,invasion,and the EMT process.Specifically,HOXA10-AS siRNA transfection significantly reduced the migratory capacity of A172 cells by 50.5%and U251 cells by 61.4%,as well as their invasive capacities by 33.8%and 58.5%,respectively(all p<0.05).HOXA10-AS acts as anmiR-99a-3p sponge,and pathway analysis identified processes linked to tumorigenesis andmetastasis,alongwith nine hub genes.HOXA10-AS upregulates the expression of integrin subunit beta 5(ITGB5)through a competing endogenous RNAmechanism.Thereduced tumorigenic behavior of glioblastoma cells due toHOXA10-AS knockdown can be rescued by ITGB5 overexpression ormiR-99a-3p inhibitor.Conclusion:These results indicate thatHOXA10-AS promotes tumorigenic behavior in glioblastoma cells by regulating the EMT-like process and functioning as an miR-99a-3p sponge to modulate ITGB5 levels,providing insights into glioblastoma development and potential therapeutic targets.展开更多
BACKGROUND Degenerative disc disease(DDD)is characterized by the loss of nucleus pulposus cells(NPCs).Inducing differentiation of bone marrow mesenchymal stem cells(MSCs)into NPCs has emerged as a novel therapeutic st...BACKGROUND Degenerative disc disease(DDD)is characterized by the loss of nucleus pulposus cells(NPCs).Inducing differentiation of bone marrow mesenchymal stem cells(MSCs)into NPCs has emerged as a novel therapeutic strategy for DDD.However,the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.AIM To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.METHODS In vitro,the effects of miR-365 on MSC proliferation,apoptosis,and differentiation were assessed by cell counting kit-8 assay,flow cytometry,and quantitative realtime polymerase chain reaction(qRT-PCR).In vivo,the expression levels of miR-365,HIF-1α,Sox9,Kdm6a,and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.RESULTS In vitro,miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis.The expression levels of glycosaminoglycans,proteoglycan,and type 2 collagen were significantly increased with miR-365 ectopic expression.In vivo,the expression levels of miR-365,HIF-1α,Sox9,and Kdm6a were significantly increased,whereas HOXA9 was remarkably decreased.Mechanically,miR-365 inhibited HOXA9 expression by directly binding to its 3’untranslated region.HOXA9 could inhibit HIF-1αexpression by binding to the Hif-1αpromoter,thereby affecting the expression levels of Sox9 and Kdm6a.Moreover,HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.CONCLUSION miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1αand may serve as a potential target for DDD treatment.展开更多
基金supported by the National Natural Science Foundation of China(No.82001243).
文摘Objectives:Glioblastoma is a prevalent malignant brain tumor,and the actions of the long non-coding RNA HOXA10-AS in its invasion and migration remain unclear.Here,the function of HOXA10-AS in glioblastoma cell invasion and migration and associated mechanisms were investigated.Methods:HOXA10-AS was knocked down in glioblastoma cells,and Transwell and wound healing assays were conducted to elucidate its impacts on cell invasion and migration.Western blotting and quantitative reverse transcription polymerase chain reaction(qRTPCR)assessed HOXA10-AS’s impact on the epithelial-mesenchymal transition(EMT).Microarray analysis identified differentially expressed genes,complemented by bioinformatics approaches to explore potentialmolecular participants and pathways.Rescue experiments validated our findings.Results:HOXA10-AS knockdown significantly inhibits glioblastoma cell migration,invasion,and the EMT process.Specifically,HOXA10-AS siRNA transfection significantly reduced the migratory capacity of A172 cells by 50.5%and U251 cells by 61.4%,as well as their invasive capacities by 33.8%and 58.5%,respectively(all p<0.05).HOXA10-AS acts as anmiR-99a-3p sponge,and pathway analysis identified processes linked to tumorigenesis andmetastasis,alongwith nine hub genes.HOXA10-AS upregulates the expression of integrin subunit beta 5(ITGB5)through a competing endogenous RNAmechanism.Thereduced tumorigenic behavior of glioblastoma cells due toHOXA10-AS knockdown can be rescued by ITGB5 overexpression ormiR-99a-3p inhibitor.Conclusion:These results indicate thatHOXA10-AS promotes tumorigenic behavior in glioblastoma cells by regulating the EMT-like process and functioning as an miR-99a-3p sponge to modulate ITGB5 levels,providing insights into glioblastoma development and potential therapeutic targets.
基金Supported by Academic Subject Boosting Plan in Shanghai Fourth People's Hospital Affiliated to Tongji University School of Medicine,No.SY-XKZT-2019-3002Academic Project from the Health Commission of Hongkou District,Shanghai,No.2202-25Clinical Key Specialty Construction Unit from The Health Commission of Hongkou District,Shanghai,No.HKLCZD2024B03.
文摘BACKGROUND Degenerative disc disease(DDD)is characterized by the loss of nucleus pulposus cells(NPCs).Inducing differentiation of bone marrow mesenchymal stem cells(MSCs)into NPCs has emerged as a novel therapeutic strategy for DDD.However,the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.AIM To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.METHODS In vitro,the effects of miR-365 on MSC proliferation,apoptosis,and differentiation were assessed by cell counting kit-8 assay,flow cytometry,and quantitative realtime polymerase chain reaction(qRT-PCR).In vivo,the expression levels of miR-365,HIF-1α,Sox9,Kdm6a,and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.RESULTS In vitro,miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis.The expression levels of glycosaminoglycans,proteoglycan,and type 2 collagen were significantly increased with miR-365 ectopic expression.In vivo,the expression levels of miR-365,HIF-1α,Sox9,and Kdm6a were significantly increased,whereas HOXA9 was remarkably decreased.Mechanically,miR-365 inhibited HOXA9 expression by directly binding to its 3’untranslated region.HOXA9 could inhibit HIF-1αexpression by binding to the Hif-1αpromoter,thereby affecting the expression levels of Sox9 and Kdm6a.Moreover,HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.CONCLUSION miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1αand may serve as a potential target for DDD treatment.
