体外转染PTEN抑制胆管癌QBC939细胞生长及下调mTOR表达的研究
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刘民锋

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Investigation of the inhibition of the cell growth and down-regulation of mTOR in the cholangiocarcinoma QBC939 cells transfected with plasmid PTEN in vitro
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    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QBC939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QBC939;用Western 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QBC939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。

    Abstract:

    Abstract: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 mTOR are closely related through phosphorylation of AKT.

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刘民锋,徐立宁,左石,罗剑,郭伟,董泾青,邹声泉.体外转染PTEN抑制胆管癌QBC939细胞生长及下调mTOR表达的研究[J].中国普通外科杂志,2006,15(3):7-184.
DOI:10.7659/j. issn.1005-6947.2006.03.007

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  • 在线发布日期: 2006-03-25