PI3K/Akt通路在肝癌细胞迁移和侵袭中的作用

Role of PI3K/Akt in migration and invasion of hepatocellular carcinoma cells

目的探讨PI3K/Akt信号通路中3-磷酸肌醇-依赖性激酶1(PDK1)、10号染色体上的磷酸酶和张力蛋白同源蛋白(PTEN)、蛋白激酶B(Akt)表达对肝癌细胞迁移和侵袭的作用。方法常规培养肝癌SMMC-7721细胞,将细胞分为对照组、PDK1抑制剂组、Akt激动剂组、PTEN抑制剂组。对照组加入RPMI-1640培养基,PDK1抑制剂组加入PDK1的特异性抑制剂OSU03012,Akt激动剂组加入Akt特异性激动剂SC79,PTEN抑制剂组加入PTEN特异性抑制剂VO-Ohpic。采用细胞划痕实验检测各组细胞迁移能力,Transwell小室检测细胞侵袭能力,Western blotting法检测PDK1基因相关蛋白PDK1、p-PDK1、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)及PTEN、Akt蛋白表达。结果与对照组比较,PDK1抑制剂组细胞迁移率减少,Akt激动剂组和PTEN抑制剂组细胞迁移率增加(P<0.05或<0.01)。与对照组比较,PDK1抑制剂组穿膜细胞数减少,Akt激动剂组和PTEN抑制剂组穿膜细胞数增加(P均<0.01)。与对照组比较,PDK1抑制剂组PDK1、p-PDK1表达降低,E-cadherin表达升高,Vimentin表达降低;Akt激动剂组Akt表达升高,PTEN表达降低;PTEN抑制剂组PTEN表达降低,Akt表达升高(P<0.05或<0.01)。结论抑制PI3K/Akt信号通路中PDK1表达可以抑制肝癌细胞迁移和侵袭,促进Akt和抑制PTEN表达可以促进肝癌细胞迁移和侵袭。

Objective To investigate the effects of 3-phosphoinositide-dependent protein kinase-1（ PDK1）,phosphatase and tensin homolog deleted on chromosome 10（ PTEN）,and protein kinase B（ Akt） expression in the PI3 K/Akt signaling pathway on the migration and invasion of hepatoma cells. Methods Liver cancer SMMC-7721 cells were routinely cultured and divided into the control group,PDK1 inhibitor group,Akt agonist group,and PTEN inhibitor group. The cells in the control group were cultured in RPMI-1640 medium,the specific inhibitor of PDK1 OSU03012 was added into the PDK1 inhibitor group,the Akt specific agonist SC79 was added to the Akt agonist group,and the PTEN specific inhibitor VO-Ohpic was added to the PTEN inhibitor group. Cell scratch assay was used to detect the migration ability of cells in each group. Transwell was used to detect the cell invasion ability. Western blotting was used to detect expression of proteins PDK1,E-cadherin,Vimentin,PTEN,and Akt. Results Compared with the control group,the mobility rate of PDK1 inhibitor group decreased,but the mobility rates of the Akt agonist group and PTEN inhibitor group increased（ P〈0. 05 or P〈0. 01）. The number of transmembrane cells in the PDK1 inhibitor group decreased（ P〈0. 01）,the number of transmembrane cells in the Akt agonist group increased,and the number of transmembrane cells in the PTEN inhibitor group increased（ all P〈0. 01）. Compared with the control group,the protein expression of PDK1 and p-PDK1 decreased,E-cadherin expression increased,and Vimentin expression decreased in the PDK1 inhibitor group; the expression of Akt increased and PTEN expression decreased in the Akt agonist group; PTEN expression decreased and the Akt expression increased in the PTEN inhibitor group（ P〈0. 05 or P〈0. 01）. Conclusion Inhibiting the expression of PDK1 in the PI3 K/Akt signaling pathway can inhibit the migration and invasion of liver cancer cells,and promoting expression of Akt and inhibiting expression of PTEN can promote the migration and invasion of liver cancer cells.