The Effect Of P38δ On Hela Cell Apoptosis

Objective:The main aim of this study is to investigate the expression level of p38δ in cervical cancer cells and Hela,the effect of p38δ on Hela apoptosis induced by hypoxia and cisplatin and the mechanism of this process,and to provide theoretical basis and new idea for the further understand and clinical application of cervical cancer.Methods : 1 Western blot analysis was used to detect the expression level of p38δ in cervical cancer cells and Hela cell line;2 Plasmid transfection was used to overly express p38δ in Hela;3 The apoptosis of Hela tansfected or not p38δ were detected by annexin V-FITC/PI double labeling;4 Hela cell line stably expressing p38δ was established by lentivirus;5 Hela cell line stably expressing p38δ were confirmed by western blot and immunofluorescence staining;6 Annexin V-FITC/PI double labeling were used to detect the apoptosis of Hela stably expressing p38δ or not induced by hypoxia and cisplatin;7 The phosphorylation levels of p38δ were detected by immunoprecipitation and western blot;8 The effect of p38δkinase-dead mutant on Hela apoptosis was detected by transient transfection p38δ(WT or K54R)and annexin V-FITC/PI double labeling;9 Western blot were used to detect the expression levels of apoptosis related proteins in Hela cell line stably expressing p38δ or not induced by hypoxia and cisplatin.Result:1 The expression level of p38δ was significantly down-regulated in cervical cancer tissue and Hela,compared to normal cervical tissue and nontumor cell lines,respectively(all p<0.05);2 Overexpression ofp38δ in Hela induced apoptosis(p<0.05);3 Compared to control Hela cell line,stably expressing p38δ Hela cell line were more sensitive to apoptosis induced by hypoxia and cisplatin(all p<0.05);4 Both hypoxia and cisplatin induced the phosphorylation of p38δ in Hela cell(all p<0.05);5 Kinase-dead mutant of p38δ decreased the level of apoptosis in Hela cell compared to wild-type p38δ(p<0.05);6 The expression levels of p53 and Bax in Hela cell line stably expressing p38δinduced by hypoxia and cisplatin were significantly higher than those in control Hela cell line,while the expression level of Bcl-2 in the former was lower than that in the later(all p<0.05).Conclusion:1 p38δ are down-regulated in cervical cell tissue and Hela cell;2 p38δ promotes the sensitivity of Hela cell to apoptosis induced by hypoxia and cisplatin,which is dependent on kinase activity of p38δ and the protein-expression regulation of Bcl-2、Bax and p53.