ABSTRACT
Objective:To observe the localization of chicken infectious anemia virus VP3 gene in normal cells and breast cancer cells in different times and its apoptosis-inducing effect.Methods: The fundamental cloning method,inserting the VP3 gene of chicken anemia virus into the eukaryotic expression vector pEGFP-C1 was used.Then,the positive recombinant containing VP3 gene pEGFP-C1-VP3 was transfected into human breast cancer cell line MCF-7 and mouse fibroblasts L929 by FuGENE(R)6 transfection reagent in vitro respectively.After 24 hours,48 hours and 72 hours,fluorescence microscope was used to observe the distribution of VP3 in cells and the rate of apoptosis was studied on the treated MCF-7 cells by FCM(Flow Cytometry).Results: The recombinant plasmid pEGFP-C1-VP3 could be localized in the nuclei of breast cancer cells,which showed the typical nuclear changes in different stages of apoptosis.In L929 cells,pEGFP-C1-VP3 underwent a process of migration from the nucleus to the cytoplasm,which didn′t induce apoptosis of L929 cells.Conclusion: VP3 located in the nucleus of MCF-7 breast cancer cells,which can led to cancer cell death by inducing apoptosis,and the apoptosis rate was higher than the control group with time dependence.VP3 located in the cytoplasm of normal cells,and didn′t induce apoptosis.