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miR-125a-5p 通过靶向 GAB2 抑制乳腺癌细胞的增殖和侵袭并诱导细胞凋亡。

MiR-125a-5p inhibits the proliferation and invasion of breast cancer cells and induces apoptosis by targeting GAB2.

机构信息

Department of Thyroid and Breast Surgery, The First Hospital of Shijiazhuang, Shijiazhuang 050011, Hebei, China.

Department of General Surgery, People's Hospital of Haiyan, Jiaxing 314300, Zhejiang, China.

出版信息

Math Biosci Eng. 2019 Jul 29;16(6):6923-6933. doi: 10.3934/mbe.2019347.

Abstract

To investigate whether miR-125a-5p can inhibit the proliferation and invasion of breast cancer cells and induce apoptosis by targeting GAB2. qRT-PCR was used to detect the expression of miR-125a-5p in normal mammary epithelial cells and breast cancer cell lines; The miR-125a-5p overexpression plasmid was transiently transfected into MDA-MB-157 cells, and the proliferation, invasion and apoptosis of breast cancer cells were detected by CCK8 kit, Transwell chamber and flow cytometry, respectively; Gene silencing was used to knock down GAB2 gene in MDA-MB-157 cells, and the changes of proliferation, invasion, apoptosis and apoptosis-related proteins in breast cancer cells were detected by CCK8 kit, Transwell chamber, flow cytometry and western blot, respectively; The direct interaction between miR-125a-5p and GAB2 was detected by dual-luciferase reporter assay. The miR-125a-5p overexpression plasmid was transiently transfected into MDA-MB-157 cells, and the expression levels of GAB2 and apoptosis-related proteins were detected by western blot. The expression of miR-125a-5p in breast cancer cell lines, MDA-MB-157 cells, MDA-MB-361 cells and MDA-MB-415 cells, was significantly lower than that in normal breast epithelial cells, MCF-10A cells; The proliferation and invasion ability of MDA-MB-157 cells transfected with miR-125a-5p were significantly inhibited, and the apoptosis rate was significantly increased; Since GAB2 knocked down, the proliferation and invasion ability of MDA-MB-157 cells were significantly inhibited, while the apoptosis rate was significantly increased, the Bax protein expression was significantly down-regulated, and the Bcl-2 protein expression was significantly up-regulated; The dual-luciferase reporter assay demonstrated that miR-125a-5p can specifically target GAB2. Transfected with miR-125a-5p, the GAB2 protein expression and Bax protein expression were significantly down-regulated, but the Bcl-2 protein expression was significantly up-regulated. miR-125a-5p inhibits the proliferation and invasion of breast cancer cells and induces their apoptosis by negatively regulating GAB2.

摘要

为了研究 miR-125a-5p 是否可以通过靶向 GAB2 抑制乳腺癌细胞的增殖和侵袭并诱导细胞凋亡。qRT-PCR 用于检测正常乳腺上皮细胞和乳腺癌细胞系中 miR-125a-5p 的表达;瞬时转染 miR-125a-5p 过表达质粒至 MDA-MB-157 细胞,分别通过 CCK8 试剂盒、Transwell 室和流式细胞术检测乳腺癌细胞的增殖、侵袭和凋亡;使用基因沉默技术敲低 MDA-MB-157 细胞中的 GAB2 基因,分别通过 CCK8 试剂盒、Transwell 室、流式细胞术和 Western blot 检测乳腺癌细胞的增殖、侵袭、凋亡和凋亡相关蛋白的变化;通过双荧光素酶报告基因实验检测 miR-125a-5p 与 GAB2 之间的直接相互作用。瞬时转染 miR-125a-5p 过表达质粒至 MDA-MB-157 细胞,通过 Western blot 检测 GAB2 和凋亡相关蛋白的表达水平。miR-125a-5p 在乳腺癌细胞系 MDA-MB-157 细胞、MDA-MB-361 细胞和 MDA-MB-415 细胞中的表达水平明显低于正常乳腺上皮细胞 MCF-10A 细胞;转染 miR-125a-5p 的 MDA-MB-157 细胞的增殖和侵袭能力明显受到抑制,凋亡率明显升高;敲低 GAB2 后,MDA-MB-157 细胞的增殖和侵袭能力明显受到抑制,凋亡率明显升高,Bax 蛋白表达明显下调,Bcl-2 蛋白表达明显上调;双荧光素酶报告基因实验证实 miR-125a-5p 可以特异性靶向 GAB2。转染 miR-125a-5p 后,GAB2 蛋白表达和 Bax 蛋白表达明显下调,但 Bcl-2 蛋白表达明显上调。miR-125a-5p 通过负调控 GAB2 抑制乳腺癌细胞的增殖和侵袭并诱导其凋亡。

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