Sox2对膀胱癌细胞化疗耐药的影响及其分子机制的探讨  

Effect of Sox2 on Chemoresistance of Bladder Cancer Cells and Its Molecular Mechanism

作　　者：张义[1] 段良斌 宋超[2] 廖文彪[2] 喻华[1] 周琦[1] ZHANG Yi1, DUAN Liang-bin1, SONG Chao2, et al.(1. Hanchuan People＇s Hospital,Hanchuan 431600,China;2. People＇s Hospital of Wuhan University, Wuhan 430060, Chin)

机构地区：[1]汉川市人民医院,湖北汉川431600 [2]武汉大学人民医院,湖北武汉430060

出　　处：《中国肿瘤》2018年第5期387-392,共6页China Cancer

基　　金：国家自然科学基金项目(31400835)

摘　　要：[目的]探讨Sox2对膀胱癌细胞化疗耐药的影响及其分子机制。[方法]选用T24人膀胱癌细胞系,采用浓度递增方式建立顺铂耐药细胞系T24/DDP,慢病毒载体转染对应细胞,进行Sox2基因过表达或敲除,得到T24-Sox2细胞和T24/DDP-sh Sox2细胞系。q RT-PCR和Western blot检测各组细胞中Sox2与多药耐药相关蛋白（MRP）表达水平。MTT实验检测各组细胞在不同浓度顺铂作用下的吸光度值并计算IC50,绘制耐药曲线。流式细胞术检测各组细胞凋亡率,并使用q RT-PCR和Western blot检测凋亡相关蛋白的表达水平。[结果]q RT-PCR和Western blot结果证实Sox2和MRP在膀胱癌顺铂耐药细胞系中高表达,耐药曲线提示Sox2与膀胱癌的顺铂耐药存在一定关联,耐药细胞系的IC50为（15.24±1.12）μg/ml,而非耐药细胞系的IC50为（4.88±0.72）μg/ml,P〈0.05。Sox2表达上调能够抑制膀胱癌细胞凋亡,凋亡率从74.62%下降到25.84%,P〈0.05。Bax和Caspase-3在Sox2高表达的细胞中呈低表达,而Bcl-2则呈高表达。[结论]Sox2基因在顺铂耐药膀胱癌细胞系中表达水平显著高于正常非耐药膀胱癌细胞,且Sox2可能是通过抑制细胞凋亡来实现对化疗敏感性的调控。[Purpose] To investigate the effect of Sox2 on chemoresistance of bladder cancer cells and its molecular mechanism. [Methods] The cisplatin-resistant T24/DDP cell line was derived from human bladder cancer T24 cells. T24 cells and T24/DDP cells were transfected with Sox2 gene or sh RNA to obtain T24-Sox2 cells and T24/DDP-sh Sox2 cells. The expression levels of Sox2 and MRP were detected by q RT-PCR and Western blot methods. The MTT assay was used to detect the cell viability and IC50 and drug resistance curve was calculated. Flow cytometry was used to detect the apoptosis rate,and the expressions of apoptosis related genes were detected by q RTPCR and Western blot. [Results] q RT-PCR and Western blot showed that Sox2 and MRP were highly expressed in cisplatin-resistant bladder cancer cell lines. The drug resistance curve suggested that there was a correlation between Sox2 and cisplatin resistance in bladder cancer. The IC50 of T24/DDP cells was（15.24±1.12）μg/ml,whereas that of T24 cells was（4.88±0.72）μg/ml（P〈0.05）. Upregulation of Sox2 inhibited the apoptosis of bladder cancer cells,and the apoptosis rate decreased from 74.62% to 25.84%（P 0.05）. The expressions of Bax and Caspase-3 were down-regulated in T24-Sox2 cells while Bcl-2 was over-expressed. [Conclusion] Sox2 expression in cisplatin-resistant bladder cancer cells is significantly higher than its parent bladder cancer cells,which is associated with the inhibition of apoptosis.

关 键 词：SOX2 膀胱癌 细胞凋亡 顺铂 化疗耐药 

分 类 号：R737.14[医药卫生—肿瘤]