RNAi抑制PI3Kp85α蛋白活性对人乳腺癌细胞系MCF-7生长抑制的体外研究  

Inhibition of Cell Proliferation by siRNA Targeting PI3Kp85α in Breast Cancer MCF-7 Cell Line

作　　者：梅玫[1] 任玉[2] 周旋[3] 苏征[2] 祁艳斌[2] 张灏[2] 姚智[4] 蒋伶活[1] 

机构地区：[1]天津大学药物科学与技术学院,天津市300072 [2]天津医科大学基础医学研究中心 [3]天津医科大学附属肿瘤医院颈一科 [4]天津医科大学基础医学院免疫教研室

出　　处：《中国肿瘤临床》2009年第23期1360-1364,共5页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30670802);天津市应用基础与前沿技术重点项目资助(09JCZDJCI19700)~~

摘　　要：目的:探讨敲低P13Kp85α表达对人乳腺癌细胞系MCF-7细胞生长的影响和机制。方法:用靶向P13Kp85α的siRNA转染人乳腺癌细胞系MCF-7,使用Real-time PCR法鉴定转染P13Kp85α表达水平;MTT法评价P13Kp85α siRNA对乳腺癌细胞系MCF-7生长的影响;流式细胞术检测转染后细胞周期分布和凋亡;采用免疫荧光染色及western blot方法观察IA型P13K/AKT通路主要成员的表达。结果:Real-time PCR结果显示P13Kp85αsiRNA转染导致P13Kp85α表达下调;MTF结果显示P13Kp85αsiRNA转染抑制肿瘤细胞生长;流式细胞术检测可见P13Kp85α siRNA转染组细胞周期存在G_0/G_1期阻滞而且凋亡率显著高于对照组与空载体组(F=19.255,P=0.002)。结论:应用P13Kp85α siRNA转染人乳腺癌细胞系MCF-7细胞,可抑制其增殖和诱导细胞凋亡,因此P13Kp85α可以作为人乳腺癌基因治疗的候选靶点。Objective： To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods： Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85α siRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results： The expression of PI3Kp85α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion： The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

关 键 词：PI3Kp85α RNA干扰 乳腺癌 基因治疗 

分 类 号：R737.9[医药卫生—肿瘤] R979.1[医药卫生—临床医学]