重组腺病毒介导的FOXO/MDA-7对乳腺癌MDA-MB-231细胞凋亡的影响及其机制

Effect of adenovirus-mediated FOXO/MDA-7 on apoptosis of breast cancer MDA-MB-231 cells and relevant mechanism

目的探讨重组腺病毒介导的FOXO/MDA-7对乳腺癌MDA-MB-231细胞凋亡的影响及其机制。方法将MDA-MB-231细胞分为Ad-FOXO/MDA-7组、Ad-FOXO组、Ad-MDA-7组和空白对照组,采用Real-time PCR、q RTPCR和Western blot法检测各组细胞中FOXO和MDA-7基因的转录及蛋白表达水平;MTT法检测细胞的增殖活力;流式细胞术检测细胞的凋亡情况;Transwell法检测细胞的侵袭能力;Western blot法检测细胞中Ras、Raf、MEK和ERK蛋白的表达情况。结果与其他各组相比,转染Ad-FOXO/MDA-7后,MDA-MB-231细胞中FOXO和MDA-7的表达明显增强,细胞的增殖活力和侵袭能力明显被抑制(P<0.05),凋亡率明显升高,细胞中Ras、Raf、MEK和ERK蛋白的表达也明显被抑制。结论重组腺病毒Ad-FOXO/MDA-7能明显促进乳腺癌细胞的凋亡,为乳腺癌的基因治疗提供了新的思路。

Objective To investigate the effect of adenovirus-mediated FOXO / MDA-7（Ad-FOXO / MDA-7）on apoptosis of breast cancer MDA-MB-231 cells as well as the relevant mechanism. Methods MDA-MB-231 cells were divided into four groups. The cells in blank control group were untreated, with those in three test groups were treated with AdFOXO / MDA-7, Ad-FOXO and Ad-MDA-7 respectively. The gene transcription and protein expression levels of FOXO and MDA-7 in cells of various groups were determined by real-time PCR, q RT-PCR and Western blot, while the proliferation activity by MTT assay, the apoptosis by flow cytometry, the invasion ability by Transwell assay, and the expressions of Ras, Raf, MEK and ERK proteins by Western blot. Results Both the expression levels of FOXO and MDA-7 in Ad-FOXO /MDA-7 group increased significantly as compared with those in other groups, while the proliferation activity and invasion ability of cells were inhibited significantly（P 〈0. 05）, and the apoptosis rate increased significantly. However, the expressions of Ras, Raf, MEK and ERK were also inhibited significantly. Conclusion Ad-FOXO / MDA-7 significantly promoted the apoptosis of MDA-MB-231 cells, which provided a novel route for the gene therapy of breast cancer.