lncRNA CCAT1通过miR-490-3p对膀胱癌细胞化疗耐药的影响
Effect of lncRNA CCAT1 via miR-490-3p on Chemoresistance of Bladder Cancer Cells
张慧明 1乔庆东 1李志慧 1常海青1
作者信息
- 1. 453000 河南省新乡市中心医院,新乡医学院第四临床学院
- 折叠
摘要
目的 探讨lncRNA CCAT1通过miR-490-3p对膀胱癌细胞化疗耐药的影响.方法 构建膀胱癌细胞T24耐顺铂细胞株.荧光素酶报告实验检测lncRNA CCAT1与miR-490-3p的结合、miR-490-3p与KDM7A mRNA-UTR的结合.敲低lncRNA CCAT1或敲低KDM7A或过表达miR-490-3p后,检测耐顺铂细胞株的凋亡水平.结果 顺铂处理后,相对于对照膀胱癌细胞系T24,膀胱癌顺铂耐药细胞系T24的凋亡水平显著下降(P＜0.05).敲低LncRNA CCAT1后顺铂使膀胱癌顺铂耐药细胞系T24的凋亡水平显著上升(P＜0.05).过表达miR-490-3p后,相比于LncRNA CCAT1突变型,荧光素酶报告实验发现LncRNA CCAT1野生型的荧光素酶活性下降(P＜0.05).过表达miR-490-3p后,顺铂使膀胱癌顺铂耐药细胞系T24的凋亡水平显著上升(P＜0.05).过表达miR-490-3p后,相比于KDM7A-3 UTR突变型,荧光素酶报告实验发现KDM7A-3UTR野生型的荧光素酶活性下降(P＜0.05).过表达miR-490-3p后,膀胱癌顺铂耐药细胞系T24中KDM7A的mRNA和蛋白表达水平均下降(P＜0.05);敲低miR-490-3p后,膀胱癌顺铂耐药细胞系T24中KDM7A的mRNA和蛋白表达水平均上升(P＜0.05).敲低KDM7A后,顺铂使膀胱癌顺铂耐药细胞系T24的凋亡水平显著上升(P＜0.05).结论 lncRNA CCAT1/miR-490-3p/KDM7A促进了膀胱癌细胞对化疗药物的抵抗.
Abstract
Objective To explore the impact of the long non-coding RNA(lncRNA)CCAT1 through miR-490-3p on chemotherapy resistance in bladder cancer cells.Methods A cisplatin-resistant cell line derived from bladder cancer cells(T24)was established.A luciferase reporter assay was conducted to investigate the interaction between lncRNA CCAT1 and miR-490-3p,as well as miR-490-3p and KDM7A mRNA-UTR.Subsequently,the apoptosis levels in cisplatin-resistant cell lines were as-sessed following the knockdown of lncRNA CCAT1,KDM7A,or overexpression of miR-490-3p.Results The apoptosis level in the cisplatin-resistant bladder cancer cell line T24 exhibited a significant decrease after cisplatin treatment compared to the control bladder cancer cell line T24(P＜0.05).Following cisplatin treatment,the knockdown of LncRNA CCAT1 led to a notable in-crease in the apoptosis level in the cisplatin-resistant bladder cancer cell line T24(P＜0.05).Upon overexpression of miR-490-3p,a luciferase reporter assay demonstrated a decrease in luciferase activity in LncRNA CCAT1 wild type compared to LncRNA CCAT1 mutant(P＜0.05).Furthermore,overexpression of miR-490-3p resulted in a significant increase in the apoptosis level of the cisplatin-resistant bladder cancer cell line T24 after cisplatin treatment(P＜0.05).In addition,a luciferase reporter assay re-vealed reduced luciferase activity in KDM7A-3 UTR wild type compared to KDM7A-3 UTR mutant after overexpression of miR-490-3p(P＜0.05).The mRNA and protein expression levels of KDM7A in the cisplatin-resistant bladder cancer cell line T24 de-creased significantly after overexpression of miR-490-3p(P＜0.05),while they increased after knockdown of miR-490-3p(P＜0.05).Subsequent to KDM7A knockdown,cisplatin treatment significantly increased the apoptosis level in the cisplatin-resistant bladder cancer cell line T24(P＜0.05).Conclusion The lncRNA CCAT1/miR-490-3p/KDM7A axis facilitated the resistance of bladder cancer cells to chemotherapeutic drugs.
关键词
lncRNA/CCAT1/miR-490-3p/KDM7A/膀胱癌/耐药Key words
lncRNA CCAT1/miR-490-3p/KDM7A/Bladder cancer/Drug resistance引用本文复制引用
基金项目
河南省医学科技攻关计划项目(LHGJ20220992)
出版年
2024
NETL
NSTL
万方数据