敲降lncRNA UCA1降低人膀胱癌细胞系T24对吉西他滨耐药  

Knockdown of lncRNA UCA1 reduces gemcitabine resistance of human bladder cancer cell line T24

作　　者：周昌东[1] 林洋[1] 孙凯[1] 田玉新[1] ZHOU Changdong;LIN Yang;SUN Kai;TIAN Yuxin(Department of Urology,Jilin Province Cancer Hospital,Changchun 130012,China)

机构地区：[1]吉林省肿瘤医院泌尿外科,长春130012

出　　处：《基础医学与临床》2024年第8期1113-1119,共7页Basic and Clinical Medicine

基　　金：吉林省卫健委基金支持项目(2021LC126)。

摘　　要：目的探讨lncRNA UCA1对人膀胱癌细胞系T24体外对吉西他滨(GEM)耐药的影响及相关分子机制。方法用RT-qPCR检测T24细胞和T24/GEM细胞中UCA1 mRNA表达。用不同浓度GEM(0.1、1和10μmol/L)刺激T24/GEM细胞48 h,用LC3染色法观察自噬斑,Western blot检测自噬相关蛋白质表达。将UCA1-shRNA或UCA1-shRNA+pcDNA-Bcl-2转入T24/GEM细胞,用MTT法和流式细胞测量术评估细胞对GEM的敏感性;用Western blot检测p53、Bcl-2和自噬相关蛋白质的表达。结果T24/GEM细胞中UCA1的表达显著高于亲本T24细胞(P<0.05)。在0~10μmol/L浓度范围的GEM呈依赖性促进T24/GEM细胞自噬(P<0.05)。敲降UCA1后,T24/GEM细胞对GEM的敏感性增加(P<0.05),自噬能力减弱(P<0.05),p53和Bcl-2表达降低(P<0.05)。Bcl-2过表达能部分逆转UCA1-shRNA对T24/GEM细胞的GEM增敏和抑制自噬的作用(P<0.05)。结论敲降lncRNA UCA1通过抑制Bcl-2介导的自噬降低T24/GEM细胞对GEM的耐药性。Objective To investigate the in vitro effect of lncRNA UCA1 on gemcitabine(GEM)resistance of bladder cancer cell line T24 and its related molecular mechanism.Methods The mRNA expression of UCA1 in T24 cells and in T24/GEM cells was detected by RT-qPCR.The T24/GEM cells were incubated with varying concentrations(0.1,1,and 10μmol/L)of GEM for 48 hrs.LC3 staining microscopy was employed to visualize autophagic puncta,while the expression of autophagy-related proteins was assessed by Western blot.UCA1-shRNA and UCA1-shRNA+pcDNA-Bcl-2 were transferred into T24/GEM cells,the sensitivity of cells to GEM was evaluated by MTT method and flow cytometry;the expressions of p53 and Bcl-2 were detected by Western blot.Results The expression level of UCA1 in T24/GEM cells was significantly higher than that of parental T24 cells(P<0.05).The concentration of GEM in the range of 0-10μmol/L significantly induced dose-dependent autophagy in T24/GEM cells(P<0.05).Knockdown of UCA1 enhanced the sensitivity of T24/GEM cells to GEM(P<0.05),while reducing autophagy(P<0.05)and down-regulating the expression of p53 and Bcl-2(P<0.05).Over-expression of Bcl-2 partially reversed the GEM sensitization and autophagy inhibition of UCA1-shRNA in T24/GEM cells(P<0.05).Conclusions Knockdown of lncRNA UCA1 reduces GEM resistance of T24/GEM cells by inhibiting Bcl-2 mediated autophagy.

关 键 词：膀胱癌 lncRNA UCA1 BCL-2 自噬 吉西他滨 

分 类 号：R737.1[医药卫生—肿瘤]