Construction of A Human Cervical Cancer Cell Line Containing the Main Capsid Protein VP5 of Stable Oncolytic Herpes Simplex Virus Type Ⅱ
This paper aims to establish a human cervical cancer HeLa VP5 cell line stably expressing the main capsid protein(VP5)of oncolytic herpes simplex virus type Ⅱ,and to lay a foundation for exploring whether VP5 protein regulates the cytotoxicity of natural killer cells(NK).To this end,pPBDP-UL19 and pSPBT plasmids were transferred into HeLa cells by using piggyBac transposition system and lipo-fectamine 3000 TM liposome transfection method;Purinomycin(Puro)was used to screen stable cell lines;Monoclonal cells were obtained by finite dilution method;Western blot was used to identify the ex-pression of VP5 protein in HeLa VP5 cell line;The expression of GFP in HeLa VP5 was detected by flow cytometry;Live cell real time monitoring method was used to compare the growth activity changes of cell lines before and after transformation.The results show that three HeLa VP5 monoclone cells were ob-tained by purinomycin resistance screening and limited dilution selection;Western blot showed that VP5 protein was expressed in three HeLa VP5 cell lines;The positive rate of GFP was 99％by flow cytometry;The live cell real time monitoring method showed that the HeLa VP5 cell line had the same growth activity as the parent cell HeLa cells.The HeLa VP5 stably transfected cell line has been successfully constructed as a target cell to evaluate whether VP5 protein regulates the cytotoxicity of natural killer cells(NK),pro-viding a theoretical basis for revealing the mechanism of oHSV2's anti-tumor effect.
oncolytic herpes simplex virus type ⅡMain capsid proteinStable cell line
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维普
