微小RNA-145 影响乳腺癌细胞系MCF-7增殖和凋亡的机制

doi:10.16098/j.issn.0529-1356.2020.01.010

摘要/Abstract

摘要：

目的探讨微小RNA-145（miR-145）参与乳腺癌MCF-7细胞增殖与凋亡的机制。方法体外培养永生化乳腺癌细胞系MCF-7细胞，并转染miR-145。分别应用Real-time PCR与Western blotting检测mRNA和蛋白水平。细胞计数试剂盒8（CCK-8）检测各组细胞增殖水平，流式细胞术检测不同处理组MCF-7细胞的凋亡水平。结果 Real-time PCR结果表明，高表达miR-145对MCF-7细胞Caspase-3、增殖细胞核抗原（PCNA）和Bcl-2的mRNA水平并无影响；而Western blotting实验结果表明，与对照组相比，转染miR-145 96 h可显著提高Caspase-3的表达，并抑制PCNA与Bcl-2的表达。CCK-8实验结果表明，过表达miR-145 72 h、96 h 后MCF-7细胞增殖率降低，差异具有统计学意义(P<0.05)。流式细胞术结果表明，过表达组MCF-7细胞的凋亡率显著高于对照组，差异具有统计学意义(P<0.05)。结论 miR-145可通过下调PCNA与Bcl-2蛋白、上调Caspase-3蛋白的表达，从而抑制MCF-7细胞增殖和促进MCF-7细胞凋亡，有望成为乳腺癌治疗的新靶点。

关键词: 乳腺癌, MCF-7细胞系, 微小RNA-145, 细胞增殖, 免疫印迹法, 人

Abstract:

Objective To explore the mechanism of microRNA-145 (miR-145) involved in proliferation and apoptosis of breast cancer MCF-7 cells. Methods The immortalized breast cancer cell line MCF-7 cells were cultured in vitro and transfected with miR-145. Real-time PCR and Western blotting were used to detect mRNA and protein levels, respectively. The proliferation level of each group was detected by cell counting kit-8 (CCK-8) method , and the apoptosis level of MCF-7 cells in different treatment groups was detected by flow cytometer. Results Real-time PCR result showed that miR-145 did not affect Caspase-3, proliferating cell nuclear antigen (PCNA) and B cell lymphoma factor 2 (Bcl-2) mRNA levels in MCF-7 cells. Western blotting analysis showed that compared with the control group, transfection of miR-145 for 96 hours significantly increased the expression of Caspase-3 and inhibited the expression of PCNA and Bcl-2. The result of CCK-8 assay showed that the proliferation rate of MCF-7 cells was decreased after overexpression of miR-145 for 72 hours and 96 hours (P<0.05). The result of flow cytometer showed that the apoptosis rate of MCF-7 cells in overexpression group was significantly higher than that in the control group (P<0.05). Conclusion MiR-145 can inhibit the proliferation and promote the apoptosis of MCF-7 cells by down-regulating PCNA and Bcl-2 and up-regulating the expression of Caspase-3, which may be a new target for breast cancer treatment.

Key words: Breast cancer, MCF-7 cell line, MicroRNA-145, Cell proliferation, Western blotting, Human

中图分类号:

R736.8

齐玉林韩慧芳张海新. 微小RNA-145 影响乳腺癌细胞系MCF-7增殖和凋亡的机制[J]. 解剖学报, 2020, 51(1): 58-61.

QI Yu-lin HAN Hui-fang ZHANG Hai-xin. Mechanism of microRAN-145 on proliferation and apoptosis of breast cancer cell line MCF-7#br#[J]. Acta Anatomica Sinica, 2020, 51(1): 58-61.

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