Effect Of UHRF2 In DNA Damage Response Induced By Hydroxyurea

Objective To investigate the relationship between UHRF2 and DNA damage induced by hydroxyurea. To determine the association of UHRF2 with p53,p21 and acetylated H3K9 in DNA damage response, further to explore the function of UHRF2.Methods HEK293 cells cultured in vitro treated with hydroxyurea for different time to construct cell models of DNA damage; p CMV-Flag-NIRF and UHRF2-siRNA were transfected into HEK293 cells, RT-PCR and western blot were performed to determine the expression of UHRF2; comet assay was used to test the association between UHRF2 and DNA damage; The expression of UHRF2 in DNA damage response induced by hydroxyurea was detected by western blot; The effects of UHRF2 on p53, p21 and H3K9 acetylation in DNA damage response were detected by western blot; CCK8 and flow cytometry were used to evaluate cell proliferation and cycle.Results compared with control group, HEK293 cells of experiment group treated with 2.5mmol/L hydroxyurea 12 h later, obvious DNA damage was detected, and no significant differences compared with 24 h treatment group;After pCMV-FLAG-UHRF2 and UHRF2-siRNA were transfected into HEK293 cells, the expression of UHRF2 changed Significantly(p<0.01); the level of UHRF2 increased in DNA damage response induced by hydroxyurea; after UHRF2 was up-regulated, the tail length of comet and cell proliferation dramatically decreased(p<0.01). p53, p21, H3K9 acetylation and cells in G1 stage increased(P<0.05); knockdown of UHRF2 decreased the expression of p53, p21 and H3K9 acetylation in DNA damage response induced by hydroxyurea(p<0.05);compared with control group, cell proliferation increased and no significant changes in cell counts in G1 stage.Conclusion UHRF2 can promote DNA damage repair induced by hydroxyurea and the expression of p53, p21, acetylation of H3K9, and more significant influence on P21. UHRF2 can inhibit cell proliferation and elicit G1/S arrest in DNA damage response of HEK293 cells induced by hydroxyurea. The molecular mechanism of UHRF2 regulating DNA damage response need further studying。...