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p38δ对宫颈癌HeLa细胞顺铂敏感性的影响
沈祥丽,张勇,柳怡,张勇武
0
(成都市锦江区妇幼保健院妇产科;西南医科大学临床医学院;绵阳市中心医院妇产科)
摘要:
【摘要】 目的 探讨p38δ对HeLa细胞顺铂敏感性的影响。方法 采用细胞转染方法在HeLa细胞中过表达p38δ蛋白;采用Annexin VFITC/PI双染色检测细胞凋亡水平;采用蛋白质印迹法检测蛋白表达水平;采用免疫沉淀法纯化p38δ蛋白,并采用蛋白质印迹法检测其磷酸化水平。结果 与转染对照质粒HeLa细胞相比,转染p38δ表达质粒HeLa细胞在正常培养条件下和顺铂处理条件下的细胞凋亡水平都显著上升(P均<001)。在顺铂处理条件下,与转染对照质粒HeLa细胞相比,转染p38δ表达质粒HeLa细胞Bcl2蛋白表达水平下降,Bax和p53蛋白表达水平上升(P均<001)。顺铂处理HeLa细胞中p38δ磷酸化水平显著上升(P<001)。与转染野生型p38δ(WT)HeLa细胞相比,转染激酶失活突变体p38δ(K54R)HeLa细胞在正常培养条件下和顺铂处理条件下的细胞凋亡水平都显著下降(P均<001)。结论 p38δ促进HeLa细胞凋亡并增强其对顺铂的敏感性,且p38δ对HeLa细胞凋亡的调控依赖于其蛋白激酶活性。
关键词:  HeLa细胞  p38δ  细胞凋亡  顺铂  蛋白激酶
DOI:
基金项目:
Effect of overexpression p38δ on cisplatin chemosensitivity in HeLa cells
SHEN Xiangli,ZHANG Yong,LIU Yi,ZHANG Yongwu
(Department of Gynaecology and Obstetrics, Jinjiang Maternal and Child Health Care Hospital;The Clinical Medicine of Southwest Medical University;The Clinical Medicine of Southwest Medical University; Department of Gynaecology and Obstetrics, Mianyang Central Hospital)
Abstract:
【Abstract】 Objective To investigate the effect of p38δ on HeLa apoptosis induced by cisplatin. Methods Liposomemediated DNA transfection was used to overexpress p38δ in HeLa cells. Annexin VFITC/PI double labeling was used to detect the apoptosis of HeLa cells induced by cisplatin or not. Western blot was used to detect the protein expression levels and the phosphorylation level of p38δ purified by immunoprecipitation. Results The apoptosis rate of HeLa cells transfected with p38δ plasmids were significantly higher than that of control HeLa cells both under normal conditions and cisplatin treatment. Under cisplatin treatment, both the protein expression levels of p53 and Bax in HeLa cells transfected with p38δ plasmids were significantly higher than those in control HeLa cells, while the expression level of Bcl2 protein in HeLa cells transfected with p38δ plasmids was lower than that in control HeLa cells. The phosphorylation level of p38δ in HeLa cells treated with cisplatin was significantly higher than that in controltreated cells. The apoptosis rate of HeLa cells transfected with Kinasedead mutant p38δ plasmids were sinificantly lower than that of transfected with wildtype p38δ plasmids both under normal conditions and cisplatin treatment. Conclusion p38δ promotes the sensitivity of HeLa cell to apoptosis induced by cisplatin, which is dependent on it’s kinase activity.
Key words:  HeLa cells  p38δ  Cell apoptosis  Cisplatin  Protein kinase

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