γ-干扰素对膀胱癌T24细胞增殖影响及分子机制研究

摘要

探讨γ-干扰素(interferon-γ),IFN对对膀胱癌细胞株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；RTPCR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-y (IFN-y) and its molecular mechanism. T24 bladder cancer cell lines were stimulated by IFN-y 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-yl)-2, 5-diphenyl-tetrazolium bromide) colorimetric assay. The median concentration (500 U/mL IFN-y) was then determined to stimulate the cells, and set as experiment groups, without IFN-y processing set to the control group. Cell cycles were examined by flow cytometry. The expression of hepaCAM (hepatocyte cell adhesion molecule) mR-NA was determined by RT-PCR. Western bolt was used to detected the protein expression of p21WAFI. Results showed: IFN-y inhibited the proliferation of T24 cells in time-dose dependent pattern (P<0.05). Compared with the control group, cell cycles were arrested at G