胰岛素样生长因子2引起Rh1肉瘤细胞mTOR信号的背景变化

Insulin-like growth factor-Ⅱ induces mTOR pathway change in Rh1 sarcoma cells

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。

Objective To observe the effect of insulinlike growth factor-Ⅱ（IGF-2） on the growth and the mTOR pathway of Rh1 sarcoma cells.Methods Rh1 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.Results IGF-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.Conclusion IGF-2 can gradually increase the function of S6 in the mTOR pathway,and the function of Akt（s473） is kept relatively stable.