摘要
目的探讨miR-15a在诱导乳腺癌细胞凋亡中的作用及机制。方法采用定量聚合酶链反应检测人乳腺上皮细胞株MCF-10A和乳腺癌细胞株MCF-7中miR-15a的表达水平。采用软件预测miR.15a的靶点,并通过荧光素酶报告基因系统验证。将miR-15a经脂质体法转染MCF-7细胞,采用Westernblot法检测Bcl-2蛋白的表达,并采用流式细胞术检测MCF-7细胞的凋亡率。结果miR-15a在乳腺癌细胞株MCF-7中的表达明显低于乳腺上皮细胞株MCF-10A(0.253:1,P〈0.0001)。miR-15a可显著抑制抗凋亡基因Bcl-23'-UTR荧光素酶报告基因的表达(P〈0.05)。与未转染组(对照组)相比,miR-15a转染组MCF-7细胞中Bcl-2的表达水平显著下降,凋亡明显增加(P〈0.05)。结论miR-15a作为一个潜在的抑癌基因,可通过靶向于Bcl-2诱导乳腺癌细胞MCF-7发生凋亡,其可为乳腺癌的临床治疗提供新的靶点和理论依据。
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 Bel-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-IOA cells ( 0. 253 : 1, P 〈 O. 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 transfeeted 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.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2011年第11期827-830,共4页
Chinese Journal of Oncology
基金
杭州市科技发展计划项目(20090833808)
杭州市医药卫生科技计划项目(20108010)