摘要
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0.5μg/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CCCHB-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。
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.
出处
《现代肿瘤医学》
CAS
2017年第18期2861-2865,共5页
Journal of Modern Oncology
基金
国家自然科学基金(编号:81301834
30901480)
关键词
膀胱尿路上皮癌
长链非编码RNA
GAS5
化疗耐药
多柔比星
bladder urinary tract epithelial carcinoma
long non-coding RNA
GAS5
chemotherapy resistance
doxorubicin
