RunX2的真核表达载体的构建及其对乳腺癌细胞增殖和迁移的影响

Construction of Run X2 eukaryotic expression vector and study the proliferations and migrations effective of Run X2 on breast cancer cells

目的构建人Run X2真核表达载体,观察Run X2对乳腺癌细胞增殖和迁移的影响。方法运用逆转录PCR(RT-PCR)扩增、酶切、连接等基因重组技术将人Run X2基因插入pc DNA3.1真核表达载体中;并经酶切、PCR、测序鉴定;将构建的Run X2真核表达载体瞬时转染MCF-7细胞,利用Western blot法检测Run X2蛋白表达,用MTT实验和细胞划痕实验检测其对乳腺癌细胞的生物学影响。结果构建的Run X2真核表达载体在MCF-7细胞中高表达Run X2蛋白;发现高表达Run X2的MCF-7细胞活力增加,移动能力增强。结论构建的Run X2真核表达载体能促进乳腺癌细胞的增殖和迁移。

Objective To investigate the biology effect of Run X2 gene after constructing the eukaryotic expression vector of human Run X2 in breast cancer cells. Methods By the recombinant techniques such as PCR amplification,digestion,ligation,the human Run X2 gene was inserted into the eukaryotic expression vector of pc DNA3. 1,and then it was identified by restriction enzyme digestion,RT-PCR,sequencing. Eukaryotic expression vector of human Run X2 gene was transiently transfected into MCF-7 cells. Western blot analysis was applied to detect the expression of Run X2 protein in breast cancer cells MCF-7; MTT assay and wound healing assay were used to detect the cells proliferation and invasion of MCF-7. Results Our result showed that the eukaryotic expression vector of Run X2 was highly expressed Run X2 protein in MCF-7 cells,the cell proliferation and migration abilities were enhanced in MCF-7 cells with high expression of Run X2 protein. Conclusion Run X2 constructs eukaryotic expression vector and promotes malignant behavior of breast cancer cells.