布洛芬通过PI3K/Akt/mTOR信号通路调控肝癌QGY-7703细胞迁移和侵袭的机制

南振华; 潘灵辉

doi:10.3971/j.issn.1000-8578.2016.12.005

布洛芬通过PI3K/Akt/mTOR信号通路调控肝癌QGY-7703细胞迁移和侵袭的机制

Mechanism of Ibuprofen Regulating Migration and Invasion of Liver Cancer QGY-7703 Cells Through PI3K/Akt/mTOR Signaling Pathway

摘要

摘要:
目的探讨布洛芬通过PI3K/Akt/mTOR信号通路对肝癌QGY-7703细胞迁移和侵袭的调控机制。
方法取对数生长期人肝癌细胞QGY-7703，采用随机法分为两组，对照组（C组）:加入等容量的RPMI l640培养液；实验组（B组）:按照不同布洛芬浓度分为三个亚组:B1组250 μmol/L，B2组500 μmol/L、B3组1 000 μmol/L。各组分别作用24、48和72 h后，利用Transwell小室检测各组细胞的侵袭和迁移能力。各组分别作用48 h后，采用Real-time PCR检测各组细胞中PI3K、PTEN和MMP-9基因表达的变化；用Western blot检测各组细胞中PTEN、Akt、磷酸化Akt（p-Akt）、mTOR、磷酸化mTOR（p-mTOR）和MMP-9蛋白表达情况。
结果与C组比较，B1、B2、B3三组细胞迁移和侵袭能力均降低，具有浓度和时间的依赖性，差异有统计学意义（P<0.01）；与C组比较，B1、B2、B3三组细胞PTEN mRNA和PTEN蛋白表达明显升高，且随着布洛芬作用浓度的增加而升高，具有浓度依赖性，差异有统计学意义（P<0.05）；与C组比较，B1、B2、B3三组细胞PI3K mRNA、Akt和mTOR蛋白的表达量差异均无统计学意义（P>0.05）；与C组比较，B1、B2、B3三组细胞MMP-9 mRNA和蛋白的表达以及p-Akt、p-mTOR蛋白的表达均显著下降，且随着布洛芬作用浓度的增加而降低，具有良好的浓度依赖性，差异有统计学意义（P<0.05）。
结论布洛芬可以抑制QGY-7703细胞的迁移和侵袭能力，与布洛芬对细胞PI3K/Akt/mTOR信号通路的调控有关。

Abstract:
Objective To investigate the mechanism of ibuprofen regulating the migration and invasion of liver cancer QGY-7703 cells via PI3K/Akt/mTOR signaling pathway.
Methods The liver cancer QGY-7703 cells were divided randomly into control group(C group) and experimental group(B group). Then the experimental group was divided into three subgroups cultured by ibuprofen at different concentrations (B1-3 groups). Group B1 was exposed to ibuprofen with the final concentrations of 250 μmol/L, Group B2 with 500 μmol/L ibuprofen, and Group B3 with 1 000 μmol/L ibuprofen. The control group was exposed to normovolemic RPMIl640 nutrient solution. The migration and invasion of cells with diverse incubating time in different groups were determined by Transwell after 24, 48 or 72h. Real-time PCR was used to measure the expression levels of PI3K, PTEN, MMP-9 mRNA; Western blot was used to evaluate the expression levels of PTEN, Akt, p-Akt, mTOR, p-mTOR, MMP-9 protein after being incubated for 48h.
Results Compared with C group, the abilities of migration and invasion in the B1-3 groups were significantly decreased in concentration- and time-dependent manner(P<0.01). Compared with C group, the expression levels of PTEN mRNA and protein in B1-3 groups were up-regulated in a concentration-dependent manner with statistical differences(P<0.05). Compared with C group, the expression levels of Akt, mTOR protein and PI3K mRNA had no significant difference(P>0.05). Compared with C group, the expression levels of p-Akt, p-mTOR, MMP-9 protein and MMP-9 mRNA were down-regulated in B1-3 groups, with negative correlation with ibuprofen-treated concentration(P<0.05).
Conclusion Ibuprofen could inhibit the migration and invasion of liver cancer QGY-7703 cells, which might be related to PI3K/Akt/mTOR signal pathway.

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