Insulin Stimulates Goose Liver Cell Growth by Activating PI3K-AKT-mTOR Signal Pathway
Insulin Stimulates Goose Liver Cell Growth by Activating PI3K-AKT-mTOR Signal Pathway
复制标题
胰岛素通过激活PI 3 K-AKT-mTOR信号通路促进鹅肝细胞生长
DOI：
10.1159/000438650
复制
发表时间：
2016-01-01
影响因子：
--
通讯作者：
Zeng, Xianyin
中科院分区：
文献类型：
--
作者：
Han, Chunchun;Wei, Shouhai;Zeng, Xianyin
背景/目的：最近的研究表明PI3K-Akt-mTOR信号通路在调节细胞增殖中起着重要作用，因此我们推测胰岛素可能通过PI3K-Akt-mTOR信号通路影响鹅肝细胞的生长。针对肝细胞在体外和体内的生理状态不同的特点，建立了简化的体外细胞模型。方法：分离鹅原代肝细胞，分别加入不添加胰岛素、胰岛素或PI3K-Akt-mTOR途径抑制剂或与葡萄糖、PI3K-Akt-mTOR途径抑制剂共同培养，采用BrdU掺入法和流式细胞仪检测细胞DNA合成和细胞周期分析，实时荧光定量RT-PCR、ELISA法和Western印迹法检测细胞周期相关因子的mRNA和蛋白表达。结果：首次发现胰岛素能明显增加鹅原代肝细胞DNA合成、细胞增殖相关因子的mRNA水平和蛋白质含量。同时，胰岛素能明显增加PI3K-Akt-mTOR通路相关因子的mRNA水平和蛋白含量。而PI3K-Akt-mTOR通路阻断剂LY294002、雷帕霉素或NVP-BEZ235可显著降低胰岛素对细胞增殖的上调作用。结论：PI3K-Akt-mTOR通路在胰岛素调节的鹅肝细胞增殖中起重要作用。(C)2016年作者(S)，S.Karger AG，巴塞尔出版
Background/Aims: Recent studies have suggested a crucial role for PI3K-Akt-mTOR pathway in regulating cell proliferation, so we hypothesize that insulin acts goose hepatocellular growth by PI3K-Akt-mTOR signal pathway. Because the physiological status of liver cells in vitro is different from that in vivo, a simplified cell model in vitro was established. Methods: Goose primary hepatocytes were isolated and incubated in either no addition as a control or insulin or PI3K-Akt-mTOR pathway inhibitors or co-treatment with glucose and PI3K-Akt-mTOR pathway inhibitors; Then, cell DNA synthesis and cell cycle analysis were detected by BrdU-incorporation Assay and Flow cytometric analysis; the mRNA expression and protein expression of factors involved in the cell cycle were determined by Real-Time RT-PCR, ELISA, and western blot. Results: Here we first showed that insulin evidently increased the cell DNA synthesis, the mRNA level and protein content of factors involved in the cell proliferation of goose primary hepatocytes. Meanwhile, insulin evidently increased the mRNA level and protein content of factors involved in PI3K-Akt-mTOR pathway. However, the up-regulation of insulin on cell proliferation was decreased significantly by the inhibitors of PI3K-Akt-mTOR pathway, LY294002, rapamycin or NVP-BEZ235. Conclusion: These findings suggest that PI3K-Akt-mTOR pathway plays an essential role in insulin-regulated cell proliferation of goose hepatocyte. (C) 2016 The Author(s) Published by S. Karger AG, Basel