Akt通过磷酸化p27诱导胆管癌细胞生长  

Akt induces cell growth through phosphorylation p27 in cholangiocarcinoma cell

作　　者：王锋[1] 胡启立[2] 李健[2] 赵严[1] 刘华[1] 汤茂春[1] 夏小俊[1] 孟宏波 宋振顺[2] 徐晓蓉[1] 

机构地区：[1]同济大学附属上海市第十人民医院消化内科,上海200072 [2]同济大学附属上海市第十人民医院普外科,上海200072

出　　处：《复旦学报（医学版）》2016年第3期268-273,共6页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(81472578;81570578);上海市自然科学基金(15ZR1432800)~~

摘　　要：目的探讨Akt和p27蛋白控制胆管癌细胞生长的分子机制。方法利用突变载体、突变失活载体以及位点突变基因的转染技术,流式细胞仪检测PI3K、转染各类载体对胆管癌细胞周期的影响,免疫印迹、免疫荧光检测PI3K抑制剂、转染各类载体对P27蛋白表达及分布的影响。结果 PI3K抑制剂LY294002引起胆管癌细胞株G1期细胞周期阻滞,并引起p27蛋白入核;Akt能通过诱导p27蛋白出细胞核而解除LY294002的G1期细胞周期阻滞;Akt通过磷酸化野生型p27蛋白第157位苏氨酸逆转细胞周期阻滞。结论 Akt通过磷酸化p27第157位苏氨酸来诱导p27细胞内重定位于细胞质,促进胆管癌细胞的生长。Objective To investigate the molecular mechanisms for Akt and p27 in control of cholangiocarcinoma cell growth. Methods Mutationally activated Akt,dominate negative Akt,empty vector and phosphorylation deficient p27 were used in our study.Cholangiocarcinoma HCCC-9810 cells were treated with the above vectors,proteins or PI3 K inhibitor.The cell cycle was measured by flow cytometry.The protein level and distribution of p27 were detected by Western blot and immunofluorescence. Results LY294002,an inhibitor of PI3 K,induced G1 phase arrest in cholangiocarcinoma cell lines and increased p27 into the nucleus.Akt released LY294002-induced G1 phase arrest through increasing cytoplasmic p27 protein level.Akt reversed cells from the cell cycle arrest through wild type p27 phosphorylation at Thr157. Conclusions Akt induced p27 phosphorylation at Thr157 and p27 relocalization to an extranuclear subcellular distribution in cholangiocarcinoma cells.

关 键 词：胆管癌 P27 AKT 磷酸化 细胞周期 

分 类 号：R753.8[医药卫生—皮肤病学与性病学]