摘要
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 a key member of the Gabs family that plays important roles for cell proliferation,differentiation and migration. GAB2 has been shown to influence tumor invasion and metastasis. We demonstrated that GAB2 enhanced the migration of MCF-7 cells.Western blot showed that GAB2 was over-expressed in high metastatic MDA-MB-231 cells than in low metastatic MCF-7 cells. When MCF-7 was transfected with GAB2-expressing plasmid,increased the levels of MMP-2 and MMP-9 proteins were detected by Western blot and ELISA. Transwell assays showed that GAB2 over-expression enhanced the migration of MCF-7 cells. Western blot showed that the phosphorylation of Akt and ARK5 was significantly increased following 5 minutes exposure to 10 ng / m L EGF. These results suggested that GAB2 could be important for the migration of tumor cells,involving activated Akt-ARK5 signal pathway and up-regulation of MMP-2 and MMP-9.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2016年第1期80-84,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金青年基金项目(No.81402389)
山东省自然科学基金(No.ZR2014HL077)
山东省高等学校科技计划(No.J12LK03,J13LK03)
关键词
乳腺癌
迁移
基质金属蛋白酶-2
基质金属蛋白酶-9
信号通路
breast cancer
migration
matrix metalloproteinases-2（MMP-2）
matrix metalloproteinases 9（MMP-9）
signaling pathway
