PI3K/AKT/MTOR信号通路对肾癌A498细胞增殖、迁移及侵袭的影响研究

目的 探讨PI3K/AKT/MTOR信号通路对肾癌A498细胞增殖、迁移及侵袭的影响.方法 选用PI3K/MTOR双重阻断剂NVP-BEZ235体外处理肾癌A498细胞,浓度分别为0、100、250、500 nmol/L.48 h后采用蛋白质印迹法(Western blot)检测肾癌A498细胞中AKT和MTOR蛋白磷酸化情况;MTT法检测细胞增殖情况;细胞划痕实验检测细胞迁移能力;Transwell小室检测细胞侵袭能力;Western blot法检测细胞中E-Cadherin和PTEN蛋白表达变化.结果 肾癌A498细胞中p-AKT、MTOR、p-MTOR、E-Cadherin、PTEN的表达及细胞增殖率、迁移率、穿膜细胞数量各自在不同NVP-BEZ235浓度下比较,差异均有统计学意义(P﹤0.01).与0 nmol/L组比较,100、250、500 nmol/L的NVP-BEZ235处理肾癌A498细胞后,细胞中磷酸化蛋白p-AKT和p-MTOR的表达均下调,细胞增殖率下降,细胞迁移率下降,穿膜细胞数量减少,细胞中E-Cadherin和PTEN蛋白表达均上调,差异均有统计学意义(P﹤0.05).结论 通过阻断PI3K/AKT/MTOR信号通路可以抑制肾癌A498细胞的增殖、迁移和侵袭,可能与上调细胞中E-Cadherin和PTEN蛋白表达有关.

Effects of PI3K/AKT/MTOR signaling pathway on the proliferation,migration and invasion of human renal carcinoma cell line A498

Objective To study the influence of PI3K/AKT/MTOR signaling pathway on the proliferation, migration and invasion of human renal carcinoma cell line A498. Method Renal carcinoma cell line A498 cell were treated with PI3K/MTOR dual inhibitor NVP-BEZ235 in vitro at the concentration of 0, 100, 250, 500 nmol/L respectively. After 48 h, the phosphorylation of AKT and MTOR protein in renal carcinoma cell line A498 were detected by Western blot;the cell proliferation was measured by MTT assay;the cell migration was detected with cell scratch test;cell invasion was evaluated with Transwell assay;and the protein expression of E-Cadherin and PTEN were determined by Western blot. Result After treatment of NVP-BEZ235 at various concentrations, the expression of p-AKT, MTOR, p-MTOR, E-Cad-herin, and PTEN as well as the proliferation rate, migration rate, and the number of transmembrane cells were observed with statistically significant difference (P<0.01). Compared with the 0 nmol/L group, after renal carcinoma cell line A498 cell were treated with NVP-BEZ235 at the concentration of 100, 250, and 500 nmol/L, the phosphorylated protein expres-sion of p-AKT and p-MTOR were significantly down-regulated, and cell proliferation rate decreased, cell migration rate went down, the number of transmembrane cells was significantly reduced, while E-Cadherin and PTEN protein expres-sions were significantly up-regulated, all were of significant difference (P<0.05). Conclusion Inhibition of PI3K/AKT/MTOR signaling pathway can contain the proliferation, migration and invasion of renal carcinoma cell line A498, which may be related to the up regulation of the expression of E-Cadherin and PTEN protein.