[微小RNA-15a对乳腺癌MCF-7细胞凋亡诱导作用的影响]

[Effect of miR-15a on induction of apoptosis in breast cancer MCF-7 cells].

作者信息

Liu Yu-hua, Hong Li-quan, Yu Wang-qi, Li Xiang-yun, Zheng Xiao-yin

机构信息

Hangzhou Normal University, Hangzhou, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2011 Nov;33(11):827-30.

PMID:22335947

Abstract

OBJECTIVE

To investigate the effect and mechanism of miR-15a on the induction of apoptosis in breast cancer cells.

METHODS

To detect the expression level of miR-15a in breast cancer cell line MCF-7 cells and human mammary gland epithelial cell line MCF-10A cells by quantitative PCR. The target point of MCF-7 was predicted by software and was validated by luciferase report gene system. MiR-15a was transfected into MCF-7 cells with liposomes. The expression of Bcl-2 in MCF-7 cells was detected by Western blotting and the apoptosis rate of MCF-7 cells was detected by flow cytometry.

RESULTS

The expression level of miR-15a in MCF-7 cells was lower than that in the MCF-10A cells (0.253:1, P < 0.0001). The expression of MiR-15a was significantly inhibited by Bcl-2 (P < 0.05). Compared with the control, Bcl-2 expression was significantly decreased in the MCF-7 cells. The results of flow cytometry showed that the apoptosis rate was 13.4% in non-transfected MCF-7 cells, 15.9% in MCF-7 cells transfected with control RNA, and 31.5% in MCF-7 cells transfected with miR-15a (P < 0.05), indicating an evident induction of apoptosis in the MCF-7 cells.

CONCLUSION

miR-15a may have a potential application value in breast carcinoma biotherapy.

摘要

目的

探讨miR-15a对乳腺癌细胞凋亡诱导作用及其机制。

方法

采用定量PCR检测乳腺癌细胞系MCF-7细胞和人乳腺上皮细胞系MCF-10A细胞中miR-15a的表达水平。通过软件预测MCF-7的靶点，并经荧光素酶报告基因系统验证。用脂质体将miR-15a转染至MCF-7细胞。采用蛋白质免疫印迹法检测MCF-7细胞中Bcl-2的表达，采用流式细胞术检测MCF-7细胞的凋亡率。

结果

MCF-7细胞中miR-15a的表达水平低于MCF-10A细胞（0.253:1，P<0.0001）。Bcl-2可显著抑制miR-15a的表达（P<0.05）。与对照组相比，MCF-7细胞中Bcl-2的表达显著降低。流式细胞术结果显示，未转染的MCF-7细胞凋亡率为13.4%，转染对照RNA的MCF-7细胞凋亡率为15.9%，转染miR-15a的MCF-7细胞凋亡率为31.5%（P<0.05），表明miR-15a明显诱导了MCF-7细胞凋亡。