抑制OCT4-pg5与OCT4B表达对膀胱癌顺铂敏感性的影响  

Suppression of OCT4-pg5 and OCT4B enhanced the sensitivity of bladder cancer cells T24 to cisplatin

作　　者：周五二 杨成林 王尉 聂海波 张小明 肖远松 王葵 曹智 Zhou Wu-Er;Yang Cheng-Lin;Wang Wei;Nie Hai-Bo;Zhang Xiao-Ming;Xiao Yuan-Song;Wang Kui;Cao Zhi(Department of Urology,General Hospital of Southern Theater Command,Guangzhou 510010,China)

机构地区：[1]南部战区总医院泌尿外科,广州510010

出　　处：《解放军医学杂志》2020年第2期176-181,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81372744).

摘　　要：目的探讨八聚体结合转录因子4假基因5(OCT4-pg5)与八聚体结合转录因子4B(OCT4B)的表达对人膀胱癌细胞顺铂敏感性的影响。方法采用反转录实时定量PCR(RT-qPCR)检测OCT4-pg5、OCT4B在膀胱癌细胞及组织中的表达;选择高表达细胞株,通过RNA干扰技术下调其OCT4-pg5、OCT4B表达,细胞借此分为si-OCT4-pg5组、si-OCT4B组、si-OCT4-pg5/si-OCT4B组及对照组;采用细胞计数试剂盒8(CCK-8)检测各组细胞顺铂半数抑制浓度(IC50);RT-qPCR及Western blotting检测细胞OCT4-pg5、OCT4B表达情况;流式细胞术检测细胞凋亡及周期分布情况。结果OCT4-pg5与OCT4B在膀胱癌T24细胞及膀胱癌组织中高表达,且两者在癌组织中的表达呈正相关关系(r=0.73,P<0.01)。干扰OCT4-pg5、OCT4B表达可显著降低T24细胞的顺铂IC50,二者呈现协同效应[(29.41±3.84)μmol/L vs.(85.00±7.63)μmol/L,P<0.01]。与对照组相比,si-OCT4-pg5转染可以明显降低T24细胞OCT4B的表达水平。OCT4-pg5、OCT4B下调可明显增强T24细胞对顺铂诱导凋亡的敏感性,使S期细胞分布减少,细胞阻滞在G0/G1期。结论抑制OCT4-pg5、OCT4B表达可增加人膀胱癌T24细胞的顺铂敏感性。Objective To investigate the effects of octamer-binding transcription factor 4 pseudogene 5(OCT4-pg5)and octamer-binding transcription factor 4B(OCT4B)expression on cisplatin sensitivity of bladder cancer T24 cells.Methods The expression of OCT4-pg5 and OCT4B in bladder cancer cells and tissues were quantified by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).To examine the effect of OCT4-pg5 and OCT4B on cisplatin sensitivity,cells were transfected with OCT4-pg5 si-RNA,OCT4B si-RNA and their control si-RNA by lipofectamine 2000 followed by a 48 h cisplatin treatment.CCK8 was used to assess the half-maximal inhibitory concentration(IC50)of cisplatin in T24 cells.OCT4-pg5 and OCT4B expression were detected by RT-qPCR or Western blot.Next,flow cytometry was used to examine the effects of OCT4-pg5 and OCT4B on T24 cells apoptosis and cell-cycle distribution.Results RT-qPCR results showed that OCT4-pg5 and OCT4B were highly expressed in bladder cancer cells and tissues.Moreover,OCT4-pg5 expression was positively correlated with OCT4B expression in 23 bladder cancer tissues.Cells transfected with OCT4-pg5 si-RNA or OCT4B si-RNA showed a much lower IC50 of cisplatin,compared with cells in the control group.RT-qPCR or Western blot results showed OCT4-pg5 or OCT4B expression was significantly down-regulated after being transfected with OCT4-pg5 si-RNA or OCT4B si-RNA in cisplatin treated T24 cells.The flow cytometry results showed that OCT4-pg5 suppression or OCT4B suppression of cisplatin treated T24 cells resulted in a significant increase of cell apoptosis and marked the transition from the S phase to G0/G1 phase.Conclusions Downregulating OCT4-pg5 and OCT4B could enhance the sensitivity of bladder cancer T24 cells to cisplatin.

关 键 词：膀胱癌 OCT4-pg5 OCT4B 顺铂 药物敏感性 

分 类 号：R3.53[医药卫生—基础医学]