γ-干扰素对膀胱癌T24细胞增殖影响及分子机制研究  

Effects of Proliferation of T24 Bladder Cancer Cells Under the Stimulation of Interferon-γ and its Molecular Mechanism

作　　者：徐斌[1] 吴小候[1] 罗春丽[2] 刘安全[1] 张俊[1] 刘琪[2] 

机构地区：[1]重庆医科大学附属第一医院泌尿外科,中国重庆400016 [2]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,中国重庆400016

出　　处：《生命科学研究》2011年第5期443-448,共6页Life Science Research

摘　　要：探讨γ-干扰素(interferon-γ,IFN-γ)对膀胱癌细胞株T24增殖影响及其分子机制.5种不同终浓度(125、250、500、1 000、2 000 U/mL)重组人细胞因子IFN-γ处理人膀胱癌T24细胞,分别在24、48、72 h采用MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)法检测T24增殖抑制率;以作用浓度为500 U/mL IFN-γ作用T24细胞,并设为实验组,未经IFN-γ处理设为对照组.流式细胞仪检测T24细胞周期各阶段变化;RT-PCR检测hepaCAM(hepatocyte cell adhesion molecule)基因表达;Western blot检测p21WAF1蛋白表达.结果表明:IFN-γ抑制T24增殖呈时间剂量依赖性(P<0.05);与对照组相比,实验组经流式细胞仪检测提示细胞G0/G1期比例增高(n=5,P<0.01);RT-PCR结果提示hepaCAM基因表达水平升高(P<0.05);Western blot结果提示p21WAF1蛋白表达水平增高,有统计学意义(P<0.01).新基因hepaCAM在IFN-γ对膀胱癌细胞株T24增殖调节中可能起作用,并可能通过上调p21WAF1蛋白表达阻滞T24细胞于G0/G1期,而抑制T24增殖.To explore the effect of proliferation of T24 bladder cancer cells lines under the stimulation of interferon-γ （IFN-γ） and its molecular mechanism. T24 bladder cancer cell lines were stimulated by IFN-γ at five different final concentrations （125, 250, 500, 1 000, 2 000 U/mL） and tested at 24, 48, 72 h, respectively, the proliferation inhibition ratio of T24 was detected by MTT （3-（4, 5-Dimethylthiazol-2-γl）-2, 5-diphenyltetrazolium bromide） colorimetric assay. The median concentration （500 U/mL IFN-γ） was then determined to stimulate the cells, and set as experiment groups, without IFN-γ processing set to the control group. Cell cycles were examined by flow cytometry. The expression of hepaCAM （hepatocyte cell adhesion molecule） mRNA was determined by RT-PCR. Western bolt was used to detected the protein expression of p21^WAF1. Results showed： IFN-γ inhibited the proliferation of T24 cells in time-dose dependent pattern （P〈0.05）. Compared with the control group, cell cycles were arrested at G0/G1 phase （P〈0.01）. The hepaCAM mRNA was re-expressed slightly under the stimulation of IFN-γ （P〈0.05）. The expression of p21^WAF1 was gradually increased. The comparison between the control group was remarkably discrepant （P〈0.01）. New gene hepaCAM may play a role in IFN-γ regulating the proliferation of T24 cells through p21^WAF1 depended way and arrest-ed the cells at G0/G1 phase, then inhibited T24 proliferation.

关 键 词：膀胱癌 干扰素Γ hepaCAM基因 P21WAF1蛋白 增殖 

分 类 号：R737.14[医药卫生—肿瘤]