摘要
目的研究信号转导通路PI3K/AKT/PTEN/mTOR中抑癌基因PTEN在体外对胆管癌QBC 9 3 9细胞生长的抑制作用,及对下游mTOR表达的影响。方法用携带有野生型PTEN和突变型PTEN的真核表达载体及空载质粒转染QBC 9 3 9;用W estern blot检测转染后PTEN蛋白的表达及蛋白激酶B磷酸化的变化;流式细胞技术检测细胞周期和细胞凋亡情况,及对下游mTOR表达的影响。结果野生型PTEN转染成功后的QBC 9 3 9细胞中PTEN明显上升,磷酸化AKT明显下降,mTOR表达也明显下调;而突变型PTEN转染后的细胞中PTEN上升,但磷酸化AKT的变化不明显,mTOR的表达亦无明显变化。结论PTEN通过磷酸化AKT使之活化,进一步使下游的mTOR同步下调,继而调节肿瘤细胞周期和细胞凋亡。在PI3K/AKT/PTEN/mTOR信号转导途径中,PTEN与mTOR通过AKT的磷酸化有密切关系。
Objective To investigate the effects of the tumor suppressor gene PTEN in growing inhibition and down-regulating mTOR in cholangiocarcinoma QBC939 cells in vitro. Methods QBC939 cells were transfected with plasmids wild-type PTEN and C124S-PTEN in vitro. After transfection, the expression of the PTEN and phosphorylation of AKT and mTOR was detected by Western blot. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells. Results Compared with the control, the expression of phosphorylation AKT was decreased and mTOR were down-regulated respectively when transfected with the wild-type PTEN. However, after transfection with mutation-type PTEN, the level of PTEN in the cells by increased, but phosphorylation AKT level and mTOR expression had no significant change. Conclusions PTEN can be actived by phosphorylated AKT. Actived AKT decreased the mTOR which led to tumor cells apoptosis and regulation of the tumor cell cycle. In the pathway of signal transmission of PI3K/AKT/PTEN/ mTOR, PTEN and roTOR are closely related through phosphorylation of AKT.
出处
《中国普通外科杂志》
CAS
CSCD
2006年第3期181-184,共4页
China Journal of General Surgery
基金
国家高技术研究发展计划(863计划)资助项目(2002AA214061)
