中文摘要：

目的：构建以慢病毒质粒pCDH为载体的人Annexin A2（Anxa2）基因及其不同活性状态的突变体的重组质粒，筛选并验证稳定表达目的基因的人乳腺癌细胞MCF-7，观察不同表达水平及不同的酪氨酸磷酸化状态下Anxa2对乳腺癌细胞MCF-7增殖及侵袭能力的影响。方法：以质粒pEGFP—N3-Anxa2为模板，利用PCR定点突变技术获得Anxa2基因的不同活性状态的突变体-pEGFP—N3-Anxa2Y23A和pEGFP—N3-Anxa2Y230D。以慢病毒质粒pCDH和pEGFP—N3-Anxa2及其突变体为模板，通过亚克隆技术获得重组质粒pCDH—EGFP—Anxa2WT、pCDH—EGFP—Anxa2Y23A及pCDH—EGFP—Anxa2Y23D。利用慢病毒感染获得稳定表达Anxa2及其突变体的MCF-7细胞。分别采用MTT和克隆形成、划痕和transwell法测定MCF-7的增殖、迁移和侵袭能力。结果：表达Anxa2-WT及Anxa2-Y23D蛋白的MCF-7细胞的增殖、迁移及侵袭能力明显提高。结论：Anxa2的表达上调及其23位酪氨酸的磷酸化促进乳腺癌细胞MCF-7的增殖及侵袭。

英文摘要：

Objective： To establish the pCDH lentivirus vector carrying human Annexin A2 （Anxa2） gene and its mutants that representing its different activity patterns, screen and check the MCF-7 cells that could express the target gene consistently, and observe the influence of Anxa2 and its tyrosine 23 phosphorylation on the proliferation and invasion of breast cancer MCF-7 cells. Methods： Taking plasmid of pEGFP-N3-Anxa2 as the template, the point mutations polymerase chain reaction （PCR） was performed and the mutants： pCDH-EGFP-Anxa2TM and pCDH-EGFP-Anxa2Y23D were obtained. The plasmid and its mutants were digested by two kinds of restriction enzyme, and then directionally inserted into pCDH. After packing the lentivirus, they were infected with MCF-7 cells. The MCF-7 cells which consistently expressing target gene were screened, arid then series of experiments such as MTT, wound healing assay, cell migration and invasion assay were performed to check the change of MCF-7 cells in its proliferation and invasion. Results： It was proved that the MCF-7 cells expressing Anxa2-WT and Anxa2-Y23D had higher proliferation and invasion abilities compared with the MCF-7 and Anxa2-Y23A cells. Conclusion： Up-regulation of Anxa2 gene and its tyrosine 23 phosphorylation promotes proliferation and invasion of breast cancer MCF-7 cells.