长链非编码RNA GAS5对膀胱尿路上皮癌细胞的多柔比星耐药性研究

摘要

Objective:To research the regulation effects of long non-coding RNA(lncRNA) Growth arrest-specific 5(GAS5) to doxorubicin resistant bladder urothelial carcinoma(BUC) cells.Methods:The expression of GAS5 in BUC cells was detected by quantitive real-time PCR(qRT-PCR).The expression vector for GAS5 was transfected into T24/DOX cells,and the silenced effects were detected by qRT-PCR.The chemotherapy resistance of T24/DOX cells to doxorubicin was detected by MTT assay.Flow cytometry was applied to assess the apoptosis rate,and Western blot was used to detect the expression of Bcl2 protein.Results:The expression of GAS5 in T24/DOX cells was decreased significantly than that in J82 and T24 cells when compared with CCC-HB-2 cells.The expression vector for GAS5 could up-regulate GAS5 expression in T24/DOX cells.GAS5 over-expression inhibited resistance of T24/DOX cells to doxorubicin.Meanwhile,GAS5 could promote doxorubicin induced apoptosis in T24/DOX cells,and depress the expression of anti-apoptosis protein Bcl-2.Conclusion:Decreased lncRNA-GAS5 expression relating to the chemotherapy resistance,could inhibite doxorubicin resistance of BUC cells.%目的:研究长链非编码RNA-Growth arrest-specific 5(GAS5)对膀胱尿路上皮癌(bladder urothelial carcinoma,BUC)细胞的多柔比星耐药性的调控作用.方法:实时定量PCR法检测GAS5基因在BUC组织及细胞中的表达.将GAS5基因的表达载体转染多柔比星耐药的BUC细胞T24/DOX,实时定量PCR验证转染效果.MTT 法检测T24/DOX细胞对多柔比星的耐药性.应用0.5 μg/ml多柔比星处理T24/DOX细胞,流式细胞仪检测T24/DOX细胞的凋亡率,Western blot法检测T24/DOX细胞中Bcl2蛋白的表达.结果:与CCC-HB-2细胞相比,GAS5基因在J82和T24细胞中的表达显著下调,而其在T24/DOX细胞中表达下调更为显著.GAS5基因的表达载体能够显著上调T24/DOX细胞中GAS5基因的表达.GAS5高表达能够抑制T24/DOX细胞对多柔比星的耐药性.GAS5高表达能够显著促进多柔比星诱导的T24/DOX细胞的凋亡,并抑制抗凋亡蛋白Bcl2的表达.结论:长链非编码RNA GAS5在BUC中低表达,且与BUC的化疗耐药相关,其过表达能够抑制BUC细胞的多柔比星耐药性.