CAJ | 학술논문

目的探讨真核起始因子2α (eIF2α)的磷酸化对顺铂诱导肝癌细胞凋亡的影响. 方法将肝癌细胞随机分为对照组和实验组,在用突变载体eIF2αS51A转染和小分子抑制剂salubrinal 改变实验组细胞在顺铂作用条件下eIF2α磷酸化的基础上,用流式细胞和Western blot分别检测细胞凋亡的百分率和细胞凋亡的分子标志物.多组间比较采用方差分析,两组间比较采用t检验. 结果顺铂诱导肝癌细胞eIF2 α发生51位丝氨酸磷酸化.在突变载体转染条件下,顺铂(10μg/m1)诱导对照组和实验组SMMC-7721细胞在24 h凋亡的平均百分率分别为21.7％±1.5％和50.7％±2.1％(t=19.454,P＜0.05)；顺铂诱导对照组和实验组HepG2细胞在24h凋亡的百分率分别为21.0％±1.0％和57.3％±2.1％ (t=27.250,P＜0.05).在抑制剂作用条件下,顺铂(15 μ g/ml)诱导对照组和实验组SMMC-7721细胞在36 h凋亡的百分率分别为50.3％±2.5％和16.3％±2.1％ (t=18.031,P＜0.05)；顺铂诱导对照组和实验组HepG2细胞在36 h凋亡的百分率分别为42.0％±2.6％和12.0％±2.0％ (t=15.667,P＜0.05).同时,Western blot检测显示eIF2α的磷酸化抑制顺铂诱导的凋亡蛋白抗多聚ADP-核糖聚合酶的剪切. 结论 eIF2α的磷酸化在肝癌细胞抵抗顺铂介导的凋亡中起重要作用.
목적 탐토진핵기시인자2α (eIF2α)적린산화대순박유도간암세포조망적영향. 방법 장간암세포수궤분위대조조화실험조,재용돌변재체eIF2αS51A전염화소분자억제제salubrinal 개변실험조세포재순박작용조건하eIF2α린산화적기출상,용류식세포화Western blot분별검측세포조망적백분솔화세포조망적분자표지물.다조간비교채용방차분석,량조간비교채용t검험. 결과 순박유도간암세포eIF2 α발생51위사안산린산화.재돌변재체전염조건하,순박(10μg/m1)유도대조조화실험조SMMC-7721세포재24 h조망적평균백분솔분별위21.7％±1.5％화50.7％±2.1％(t=19.454,P＜0.05)；순박유도대조조화실험조HepG2세포재24h조망적백분솔분별위21.0％±1.0％화57.3％±2.1％ (t=27.250,P＜0.05).재억제제작용조건하,순박(15 μ g/ml)유도대조조화실험조SMMC-7721세포재36 h조망적백분솔분별위50.3％±2.5％화16.3％±2.1％ (t=18.031,P＜0.05)；순박유도대조조화실험조HepG2세포재36 h조망적백분솔분별위42.0％±2.6％화12.0％±2.0％ (t=15.667,P＜0.05).동시,Western blot검측현시eIF2α적린산화억제순박유도적조망단백항다취ADP-핵당취합매적전절. 결론 eIF2α적린산화재간암세포저항순박개도적조망중기중요작용.
Objective To investigate whether the phosphorylation (functionally inhibitive) of eukaryotic initiation factor 2-alpha (eIF2-α) affects the molecular mechanism of cisplatin-induced cellular apoptosis in human hepatocellular carcinoma (HCC).Methods The human HCC cultured cell lines SMMC-7221 and HepG2 were treated with cisplatin alone (controls; 24 h) or in combination with pre-transfection of a dominant-negative eIF2-α mutant (eIF2αS51A) or pre-exposure to an eIF2-α-specific phosphatase inhibitor (salubrinal) to decrease or increase the phosphorylation level,respectively.Changes in expression of apoptosis markers were quantitatively and qualitatively assessed by flow cytometry and westem blot analysis.The significance of differences among groups was assessed by analysis of variance testing and of differences between groups was assessed by t-test.Results Cisplatin treatment induced the appropriate functional-inhibitive phosphorylation of eIF2-α on serine 51.Cisplatin treatment (10 mg/ml) induced significant apoptosis in the eIF2αS51A pre-transfected SMMC-7721 (control:21.7 ± 1.5％ vs.50.7 ± 2.1％,t =19.454,P ＜ 0.05) and HepG2 (21.0 ± 1.0％ vs.57.3 ± 2.1％,t =27.250,<P ＜ 0.05).Salubrinal pre-treatment significantly inhibited the cisplatin (15 mg/ml)-induced apoptosis in SMMC-7721 (control:50.3 ± 2.5％ vs.16.3 ± 2.1％,t =18.031,P ＜ 0.05) and HepG2 (42.0 ± 2.6％ vs.12.0 ± 2.0％,t=15.667,P ＜ 0.05).Conclusion Phosphorylation of eIF2-α may act to inhibit cisplatin-induced apoptosis of HCC.