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-3'UTR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。
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 assessed 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 increase 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 revealed 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 decreased 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.
作者
张慧明
乔庆东
李志慧
常海青
ZHANG Huiming;QIAO Qingdong;LI Zhihui(Xinxiang Medical College,Xinxiang Central Hospital,Xinxiang,453000)
出处
《实用癌症杂志》
2024年第7期1050-1053,共4页
The Practical Journal of Cancer
基金
河南省医学科技攻关计划项目(编号:LHGJ20220992)。
