关键词: 重组质粒, 膜联蛋白A5, SiHa细胞, 增殖, 四甲基偶氮噻唑蓝法, CCK-8

Abstract: Objective To construct the recombinant plasmid pcDNA3.1-ANXA5 so as to study the affection of the overexpressed annexin A5(ANXA5) protein to the proliferation of uterine cervical carcinomal SiHa cells. Methods The ANXA5 cDNA fragment was cloned into pcDNA3.1 vector to construct the recombinant plasmid pcDNA3.1-ANXA5; The pcDNA3.1-ANXA5 plasmid and pcDNA3.1 vector were transfected into the uterine cervical carcinoma SiHa cells respectively and screened by G418. Western blotting was used to identify cell clones which overexpress ANXA5 and immunohistochemistry was applied to identify cells transfected with pcDNA3.1 vector which has a His-tag. Cells overexpressed ANXA5, cells transfected with pcDNA3.1 vector and cells without any treatment were tested by MTT and CCK-8 method to examine whether overexpressed ANXA5 protein can affect the proliferation of the SiHa cells. Results The recombinant plasmid pcDNA3.1-ANXA5 was successfully constructed and the ANXA5 gene sequence was proved correct; The proliferation of the SiHa cells without treatment and those transfected with pcDNA3.1 vector was fast and there was no difference（P>0.05）between the two kinds of cells；while the proliferation of the SiHa cells overexpressed ANXA5 was very slow. The difference from the other two kinds of cells was significant (P<0.05). Conclusion The ANXA5 protein maybe affect the

Key words: Recombinant plasmid, Annexin A5, SiHa cell, Proliferation, MTT, CCK-8