紫杉醇对PTEN不同状态的子宫内膜癌细胞株的作用及其机制探讨

The effect of taxol on endometrial cancer cells with different PTEN status

目的研究紫杉醇对不同PTEN状态子宫内膜癌细胞株中的作用及其相关机制。方法利用慢病毒载体系统,分别转染PTEN野生型的HEC-1A细胞和PTEN突变型的Ish-ikawa细胞,建立PTEN基因敲除(HEC-1-RNAi)细胞及过表达(Ishikawa-PTEN)的细胞模型。采用MTT法检测细胞增殖抑制率,流式细胞术检测细胞凋亡率,Western blot观察磷酸化AKT蛋白和Caspase-3蛋白的表达。结果紫杉醇对不同PTEN状态的细胞株均有明显的生长抑制作用,抑制率与时间、剂量成正相关。紫杉醇处理不同PTEN状态的细胞24h后,PTEN阳性细胞的IC50和磷酸化AKT蛋白水平明显低于PTEN阴性细胞,PTEN阳性细胞的凋亡率、Caspase-3的蛋白水平明显高于PTEN阴性细胞。结论 PTEN基因能增加子宫内膜癌细胞株对紫杉醇的敏感性,其可能与抑制PI3K/AKT信号通路,增强Caspase-3蛋白表达有关。

Aim To investigate the effect of taxol on endometrial cancer cells with different PTEN status and to explore the mechanisms involved in it.Methods PTEN-knockout and PTEN over-expression cell models were successfully built by lentiviral vectors.Cell viability was measured by MTT assay.Cell apoptosis was evaluated by flow cytometry analysis.Western blot was performed to detect the protein level of P-AKT and Caspase-3.Results Taxol time-dependently and dose-dependently inhibited the proliferation of endometrial cancer cells.Compared with PTEN negative cells,after taxol treating for 24h,the IC50 of taxol in PTEN positive cells was obviously lower and the apoptotic rate was significantly higher.The Western blot results showed that the P-AKT protein was downregulated and the Caspase-3 protein was upregulated in PTEN positive cells（P0.05） compared with PTEN negative cells.Conclusion PTEN can enhance the sensitivity of endometrial cancer cells to taxol through inhibiting the PI3K/AKT pathway and increasing the Caspase-3 expression.