中国医科大学学报2025,Vol.54Issue(5):448-454,7.DOI:10.12007/j.issn.0258-4646.2025.05.012
miR-10a-5p通过调控Apaf1逆转膀胱癌T24细胞顺铂耐药
miR-10a-5p reverses cisplatin resistance in bladder cancer T24 cells via regulating Apaf1
摘要
Abstract
Objective To investigate the effect of miR-10a-5p downregulation on cisplatin(DDP)resistance in bladder cancer T24 cells and elucidate the underlying molecular mechanism.Methods Quantitative real-time PCR quantified the expression of miR-10a-5p in the cisplatin-resistant bladder cancer cell line T24/DDP and its parental cell line T24.T24/DDP cells were divided into control,inhibitor NC,miR-10a-5p inhibitor,miR-10a-5p inhibitor+si-NC,and miR-10a-5p inhibitor+si-Apaf1 groups.Different concentrations of DDP were administered for 24 h.Cell proliferative activity was detected using the MTT assay,and the drug resistance index was calculated.Apop-tosis was analyzed using flow cytometry.The protein expression levels of Apaf1,cleaved caspase-9,cleaved caspase-3,and cytochrome C(Cyt C)were analyzed using Western blotting.A dual-luciferase reporter gene assay confirmed the target binding relationship between miR-10a-5p and Apaf1.Results The expression of miR-10a-5p was significantly higher in drug-resistant T24/DDP cells than in parental T24 cells(P<0.05).Compared with the inhibitor NC group,the miR-10a-5p inhibitor group exhibited increased T24/DDP cells sensitivity to DDP,decreased drug resistance index,and elevated apoptosis levels.The protein expression levels of Apaf1,cleaved caspase-9,cleaved caspase-3,and cytoplasmic Cyt C proteins were upregulated in the miR-10a-5p inhibitor group compared with the inhibitor NC group(all P<0.05).The DDP sensitivity of T24/DDP cells was reduced in the miR-10a-5p inhibitor+si-Apaf1 group compared to the miR-10a-5p inhibitor+si-NC group,accompanied by an increase in the drug resistance index and a decrease in apoptosis(all P<0.05).The double-lu-ciferase reporter gene assay results confirmed that Apaf1 was a downstream target gene regulated by miR-10a-5p.Conclusion miR-10a-5p knockdown targeting the upregulation of Apaf1reversed DDP resistance in T24/DDP cells.关键词
膀胱癌/miR-10a-5p/Apaf1/顺铂/耐药Key words
bladder cancer/miR-10a-5p/Apaf1/cisplatin/drug resistance分类
医药卫生引用本文复制引用
张鹰,李鹏,欧阳慧,刘嵩..miR-10a-5p通过调控Apaf1逆转膀胱癌T24细胞顺铂耐药[J].中国医科大学学报,2025,54(5):448-454,7.基金项目
湖南省卫生健康委卫生科研课题(D202304058812) (D202304058812)