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胰岛素样生长因子2引起Rh1肉瘤细胞mTOR信号的背景变化
黄文峰*
0
(三峡大学医学院形态学部,宜昌 443002)
摘要:
目的观察胰岛素样生长因子(IGF)2对Rh1肉瘤细胞生长活性和mTOR信号背景的影响。方法常规培养Rh1细胞,均用无血清培养液消除内源性因子影响后再用IGF-2(终浓度为10 ng/ml)刺激,72 h后用流式细胞仪检测细胞生长活性;蛋白质印迹方法观察IGF-2刺激细胞5、10、20、30、60 min后S6、Akt (s473)的动态变化。结果与对照组相比,IGF-2刺激可促进Rh1细胞存活,降低细胞凋亡率;IGF-2刺激细胞后可使S6磷酸化随着时间的延长逐渐增强;IGF-2亦导致Akt (s473)位点的磷酸化,其磷酸化状态相对稳定。结论IGF-2刺激Rh1 细胞时,mTOR信号通路中S6功能逐渐增强,Akt 功能则相对稳定。
关键词:  胰岛素样生长因子Ⅱ  Rh1肉瘤细胞; mTOR
DOI:10.3724/SP.J.1008.2011.0422
投稿时间:2010-07-21修订日期:2011-03-07
基金项目:湖北省卫生厅科研基金指导性项目(JX3C09).
Insulin-like growth factor-Ⅱ induces mTOR pathway change in Rh1 sarcoma cells
HUANG Wen-feng*
(Department of Morphology, Medical College, China Three Gorges University, Yichang 443002, Hubei, China)
Abstract:
ObjectiveTo observe the effect of insulin-like growth factor-Ⅱ (IGF-2) on the growth and the mTOR pathway of Rh1 sarcoma cells. MethodsRh1 cells were cultured routinely, and were treated with IGF-2 at a final concentration of 10 ng/ml after starving with pure RPMI 1640 medium. The growth of cells was analyzed by flow cytometry 72 h after IGF-2 treatment. The phosphorylation of S6 and Akt (s473) proteins were examined by Western blotting analysis at 5, 10, 20, 30, and 60 min after IGF-2 treatment. ResultsIGF-2 treatment promoted the survival and inhibited the apoptosis of Rh1 cells compared with the control group. IGF-2 also increased the phosphorylation of S6 in a time-dependent manner. However, the phosphorylation of Akt(s473) was relatively stable in Rh1 cells. ConclusionIGF-2 can gradually increase the function of S6 in the mTOR pathway, and the function of Akt (s473) is kept relatively stable.
Key words:  insulin like growth factor Ⅱ  Rh1 sarcoma cells  mTOR