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GRB2相关结合蛋白(GRB2-associated binding protein 2,GAB2)是Gabs家族重要成员,在细胞增殖、分化及迁移等过程中发挥重要作用。已有研究证明,GAB2与肿瘤侵袭转移相关,但目前GAB2在乳腺癌细胞侵袭中的研究少见报道。本研究证明,GAB2可提高乳腺癌MCF-7细胞迁移能力。Western印迹结果显示,GAB2蛋白在高转移细胞株MDA-MB-231中高表达;相反,在低转移细胞株MCF-7中低表达。瞬时转染结合Western印迹、酶联免疫吸附实验(ELISA)揭示,过表达GAB2明显增加MMP-2及MMP-9蛋白表达。Transwell检测显示,过表达GAB2明显增强MCF-7细胞的迁移能力。与未转染细胞比较,采用10 ng/m L表皮细胞生长因子(EGF)刺激转染细胞5 min,即可明显增强磷酸化的Akt、ARK5表达。上述结果提示,GAB2可通过参与Akt-ARK5信号通路,增加MMP-2、MMP-9蛋白表达,增强肿瘤细胞的迁移能力,这可能是GAB2促进肿瘤侵袭的部分机制。
GRB2-associated binding protein 2 (GAB2) is an important member of Gabs family and plays an important role in cell proliferation, differentiation and migration. It has been reported that GAB2 is involved in tumor invasion and metastasis, but few studies have reported GAB2 in invasion of breast cancer cells. This study demonstrates that GAB2 can enhance the migration of breast cancer MCF-7 cells. Western blot results showed that GAB2 protein was highly expressed in the highly metastatic cell line MDA-MB-231, but low in the low-metastatic cell line MCF-7. Transient transfection combined with Western blotting revealed that overexpression of GAB2 significantly increased MMP-2 and MMP-9 protein expression by ELISA. Transwell assay showed that overexpression of GAB2 significantly enhanced the ability of MCF-7 cells to migrate. Compared with untransfected cells, the expression of phosphorylated Akt and ARK5 was significantly enhanced by stimulating transfected cells with 10 ng / mL EGF for 5 min. The above results suggest that GAB2 may be involved in the Akt-ARK5 signaling pathway to increase the expression of MMP-2 and MMP-9 protein and enhance the migration of tumor cells, which may be part of the mechanism by which GAB2 promotes tumor invasion.