摘要
目的 E2F3在浸润性膀胱癌的发生、发展中起着重要作用,但是其具体调控分子机制尚不明确,本研究旨在探讨E2F3对浸润性膀胱癌细胞调控的分子机制。方法采用RNAi技术使E2F3在浸润性膀胱癌细胞系中低表达,通过蛋白质印迹法、PCR进行检测及后续实验,通过CHIP实验等观察E2F3在HT1376和TCCSUP细胞系中与信号转导及转录激活因子3(STAT3)和Ets1之间及其相关侵袭细胞因子的调节关系。结果 E2F3、STAT3和Ets1基因在高表达E2F3细胞系(HT1376和TCCSUP)中成功被沉默,CHIP测序显示,E2F3与Ets1和STAT3基因启动子结合增多只出现在过表达膀胱癌细胞系中;免疫组化和蛋白质印迹法检测结果显示,E2F3过表达细胞癌中,Ets1和STAT3基因过表达,二者呈正相关,r=0.421,P=0.023;E2F3高表达同时相关细胞因子也增加(IL-1、IL-8、TNF-α和VEGFA等),Ets1和STAT3被沉默后,上述细胞因子也相对降低,CHIP实验显示,与低表达E2F3膀胱癌相比,在高表达E2F3膀胱癌中,Ets1和STAT3启动子活性增强。结论 E2F3过表达在人膀胱癌细胞系中通过Ets1和STAT3调节免疫相关基因的表达,E2F3的过表达促进Ets1和STAT3的表达。
OBJECTIVE E2F3 plays an important role in bladder cancer, but the mechanisms of the regulation is not clear. This article is to investigate the mechanisms of E2F3 in invasive bladder cancer. METHODS E2F3 was knock- down in invasive bladder cancer ceils lines by RNAi. By the western, PCR and CHIP experiments to observe the regulation between the E2F3 and STAT3, Etsl as well as invasion related factors. RESULTS E2F3, STAT3 and Etsl were knockdown in E2F3 high-expressing cell lines （HT1376 and TCCSUP）, ChIP assays confirmed increased binding of E2F3 to the Etsl and STAT3 promoters in E2F3-overexpressing cell lines related to normal E2F3-expressing cell lines. Immunohistochemistry and Western.. Etsl and STAT3 proeins were increased in high E2F3 bladder cancer, r= 0. 421, P= 0. 023. The cytokines （IL-I,IL-8,TNF-α,VEGFA etc. ） express were increased in overexpression of E2F3 cells, knockdown of Etsl and STAT3 in these cells led to a marked decrease in the expression of most cytokines. CONCLUSIONS E2F3 overexpression in human bladder cancer cell lines modulates the expression of immune related genes through Etsl and STAT3.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2016年第12期768-774,793,共8页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(31272391)
陕西省科学技术研究发展计划(2008k09-04)
关键词
膀胱癌
E2F3
转录激活因子3
RNAI
免疫组织化学
bladder cancer
E2F3
signal transducers and activatos of transcription-3
RNAi
immunohistochemistry
