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膜联蛋白A2是一种参与调节多种病毒增殖的重要宿主蛋白。本研究通过构建过表达膜联蛋白A2的BHKAnxa2细胞系及瞬时敲低膜联蛋白A2,探讨膜联蛋白A2对脑心肌炎病毒在BHK-21细胞中增殖的影响。通过RT-PCR从BHK-21细胞中扩增膜联蛋白A2全长cDNA,测序正确后克隆入pcDNA3.1整合表达载体中;用重组载体pcDNA3.1-Anxa2转染BHK-21细胞,经G418筛选获得抗性克隆,qRT-PCR和Western blot试验证明BHKAnxa2过表达膜联蛋白A2;EMCV感染试验证明BHK-Anxa2细胞中病毒滴度高于对照组。通过siRNA降低BHK-21细胞中膜联蛋白A2表达,EMCV感染试验证明膜联蛋白A2表达下降后EMCV增殖也随之下降。以上试验结果表明鼠膜联蛋白A2表达改变可导致病毒在BHK-21细胞中的增殖发生变化,提示膜联蛋白A2与EMCV在BHK-21细胞中的增殖相关。
Annexin A2 is an important host protein involved in the regulation of multiplication of multiple viruses. In this study, we investigated the effect of annexin A2 on the proliferation of encephalomyocarditis virus (BHK-21) cells in BHKAnxa2 cell line that overexpresses Annexin A2 and transient knockdown of Annexin A2. The full length cDNA of Annexin A2 was amplified from BHK-21 cells by RT-PCR and cloned into the pcDNA3.1 expression vector. The recombinant plasmid pcDNA3.1-Anxa2 was transfected into BHK-21 cells and transfected by G418 The clones were screened by qRT-PCR and Western blot to confirm that BHKAnxa2 overexpressed Annexin A2. EMCV infection test demonstrated that the virus titer in BHK-Anxa2 cells was higher than that in control group. The expression of annexin A2 in BHK-21 cells was reduced by siRNA, and the EMCV infection test demonstrated that the proliferation of EMCV also decreased after the expression of annexin A2 was decreased. The above experimental results show that the change of the expression of Annexin A2 can lead to the change of the virus in BHK-21 cells proliferation, suggesting Annexin A2 and EMCV proliferation in BHK-21 cells.