The Research On The Identification Of Rock2Phosphorylation Sites And Its Effect On The Biological Activity Of Hepatoma Cells In UV Radiation-induced DNA Damage

Objective:To study the effect of Rock2on the biological activity of hepatoma cells throughidentification of its phosphorylation sites, and clarify the role of Rock2in theoccurrence and development process of liver cancer.Methods:1. Analyze the structure of Rock2by phosphorylation sites predict software andby bioinformatics analysis, and design phosphorylated polypeptides of the sites whichmay possibly occur phosphorylation.2. HepG2cells and MHCC97H cells are subjected to UV radiation, respectively,and then analyze the possible Rock2phosphorylation sites by Western Blot analysis.3. Apply colony and apoptosis experiments, to observe the effect of Rock2T814phosphorylation on the biological activity of hepatoma cells after DNA damage wasinduced by UV radiation.Result:1. The sites of Rock2which may occur phosphorylation include:(1) S442:C-SRKNESSQEIQ;(2) S771: C-DLKQSQQKINE;(3) T814: C-NDLKMQTQQVN.2. After UV radiation, the T814site of Rock2occurs phosphorylation.3. The cloning efficiency of MT-Rock2group is lower than WT-Rock2groupafter UV radiation.4. The apoptosis rate of MT-Rock2group is higher than WT-Rock2group afterUV radiation.Conclusion:1. The T814site of Rock2occurs phosphorylation after suffered UV radiation. 2. Rock2can repair the UV radiation-induced DNA damage after the T814phosphorylation.