Study on the influence of HPV16 E6-295T/G and E6-350T/G variation sites on the protein expression of IFNκ、MHC-Ⅰ、STAT1 and IRF9
Objective To explore the effect of variation sites E6-295T/G and E6-350T/G on the protein expression of IFNκ、MHC-Ⅰ、STAT1 and IRF9.Methods Cervical cancer cells C33A were transfected with eukaryotic expression vectors 295G/350G-GV230,295T/350G-GV230,295T/350T-GV230 as the experimental group and NC-GV230 as the control group.Immunohistochemistry and Western blot methods were used to detect the effects of HPV16 E6 295T/G and 350T/G variation sites on IFNκ、MHC-Ⅰ、STAT1 and IRF9.IBM SPSS Statistics 26 software was used for statistical analysis of the experimental results.P<0.05 was considered statistically significant.Immunohistochemical method was used to quantitatively count positive cells by image J software.Results The results of Western blot showed that the expression of IFNκ was decreased in the HPV16 E6-295G/350G variant group compared with the HPV16 E6-295T/350T prototype group and HPV16 E6-295T/350G(P<0.05,P<0.01).The expression of MHC-I in the HPV16 E6-295G/350G variants was significantly lower than that of HPV16 E6-295T/350T prototype group and HPV16 E6-295T/350G(P<0.05).The expression of STAT1 was significantly lower in HPV16 E6-295G/350G variant group than in HPV16 E6-295T/350T pro-totype group and HPV16 E6-295T/350G(P<0.01,P<0.05).IRF9 in HPV16 E6-295G/350G was significantly decreased than that of HPV16 E6-295T/350T prototype group and HPV16 E6-295T/350G(P<0.05,P<0.01).Conclusion HPV16 E6 protein can re-duce the expression of IFNκ,MHC-Ⅰ,STAT1 and IRF9,and E6 gene 295G/350G variation has the strongest effect on reducing the expression of these molecules.
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