Li-Bing Wang1, Liang Feng1, Jing He1, Bo Liu1, Jian-Guang Sun2. 1. Department of Thyroid and Breast Surgery, The First Hospital of Shijiazhuang, Shijiazhuang 050011, Hebei, China. 2. Department of General Surgery, People's Hospital of Haiyan, Jiaxing 314300, Zhejiang, China.
Abstract
Objective: To investigate whether miR-125a-5p can inhibit the proliferation and invasion of breast cancer cells and induce apoptosis by targeting GAB2. Methods: 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. Results: 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. Conclusion: miR-125a-5p inhibits the proliferation and invasion of breast cancer cells and induces their apoptosis by negatively regulating GAB2.
Objective: To investigate whether miR-125a-5p can inhibit the proliferation and invasion of breast cancer cells and induce apoptosis by targeting GAB2. Methods: 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. Results: 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. Conclusion:miR-125a-5p inhibits the proliferation and invasion of breast cancer cells and induces their apoptosis by negatively regulating GAB2.
Entities:
Keywords:
Breast cancer ; GAB2 ; apoptosis ; invasion ; miR-125a-5p ; proliferation