沉默FRAT1基因对结肠癌HT-29细胞增殖与凋亡的影响及其机制研究

目的 探讨沉默FRAT1基因对结肠癌HT-29细胞增殖、凋亡的影响及其可能分子机制.方法 将成功转染FRAT1基因RNA干扰序列的人结肠癌HT-29细胞扩大培养,MTT比色法检测细胞增殖,双染流式细胞术检测细胞凋亡率,单染流式细胞术分析细胞周期,免疫荧光法激光共聚焦显微镜下观察β-catenin蛋白变化.结果 转染组细胞较对照组细胞生长明显减缓（P＜0.01）,凋亡率明显增加（P＜0.01）,细胞周期出现G0/G1期阻滞,差异显著（P＜0.01）,胞质β-catenin蛋白表达明显下调.结论 FRAT1基因RNA干扰序列可以有效诱导结肠癌HT-29细胞凋亡和细胞周期阻滞,抑制细胞增殖,其机制可能通过下调β-catenin表达实现.

Effect of FRAT1 gene Silencing on proliferation and apoptosis of Colon cancer HT-29 cells and possible relevant mechanism

Objective To investigate the influence of FRAT1 gene on the proliferation and apoptosis of human colon cancer cell line HT-29 cells,and explore its underlying molecule mechanism. Methods Flow cytometry with Annexin V-FITC/PI double staining was employed to detect cell apoptosis,and the changes in the cell cycle and proliferation of the cells were examined with flow cytometry with PI staining and MTT colorimetric assay, respectively. β-catenin pro- tein localization was detected by indirect immunofluorescence. Results FRAT1 siRNA could restrain HT-29 cell prolif- eration,increase G0/G1 phase cell ratio,induce tumor cell apoptosis（P〈0.01）. The expression of β-catenin at the cyto- plasm was down regulated and inhibit the translocation into nucleus,Conclusion FRAT1 siRNA effectively inhibits the expression of HT-29 cells, and suppresses cell growth and improves cell apoptosis by up regulating the expressions of β- catenin.