miR-10a-5p reverses cisplatin resistance in bladder cancer T24 cells via regulating Apaf1
10.12007/j.issn.0258-4646.2025.05.012
- VernacularTitle:miR-10a-5p通过调控Apaf1逆转膀胱癌T24细胞顺铂耐药
- Author:
Ying ZHANG
1
;
Peng LI
1
;
Hui OUYANG
1
;
Song LIU
1
Author Information
1. 南华大学附属第二医院重症医学科,湖南 衡阳 421001;南华大学衡阳医学院,湖南 衡阳 421001
- Publication Type:Journal Article
- Keywords:
bladder cancer;
miR-10a-5p;
Apaf1;
cisplatin;
drug resistance
- From:
Journal of China Medical University
2025;54(5):448-454
- CountryChina
- Language:Chinese
-
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.
