Comparing the regulation effects on expression and phosphorylation of nuclear factor NF-κB p65 by HPV18E6 protein and protein kinase R

Abstract

Objective To Compare the regulation effects on expression and phosphorylation of nuclear factor NF-κB p65 by human papilloma virus 18 subtype E6 (HPV18E6) protein and protein kinase R(PKR). Methods Constructed two recombinant plasmid vectors which contained shRNA interfering sequence aiming at the targets of HPV18E6 oncogene and PKR gene, and transfected them respectively into HeLa cell, the expression of mRNA of HPV18E6, PKR and NF-κB p65 were determined with real-time quantitative PCR, the expression of protein HPV18E6, PKR, NF-κB p65 and phosphating PKR(P-PKR), phosphating NF-κB p65(P-NF-κB p65)were determined with Western Blot. The expression differences of these indicators among every group were compared with one-factor analysis of variance and Newman-keuls-q test of statistical software of SPSS15.0 edition. Results Based on interfering the expression of HPV18E6 oncogene, the expression of PKR mRNA and protein and phosphating protein in pSilencer-RNAi-E6 transfection group were higher than those in pSilencer-NC transfection group and blank control group(P 0.05). Conclusions HPV18E6 protein was the main regulator of expression and phosphorylation of nuclear factor NF-κB p65 in HeLa cell. The expression and phosphorylation of PKR were both suppressed, so PKR regulated NF-κB p65 weakly in HeLa cell. Key words: Nuclear factor; Papilloma virus, human; Protein kinases; HeLa cells