黏着斑激酶表达下调对肝癌细胞黏附迁移侵袭行为的影响

Effect of down-regulation of focal adhesion kinase on adhesion, migration and invasion of liver cancer cells

目的研究下调黏着斑激酶(FAK)表达对人肝癌细胞HCC-LM3黏附迁移侵袭行为的影响及可能涉及的机制。方法根据处理方法不同,将人肝癌细胞HCC-LM3分为未处理组(肿瘤细胞未经处理)、对照组(肿瘤细胞转染空载体,FAK表达未改变)和FAK-sh RNA组(肿瘤细胞稳定低表达FAK)。分别采用细胞黏附实验、划痕实验、Transwell实验检测3组肝癌细胞的黏附、迁移、侵袭能力,Western blotting检测细胞黏附侵袭相关蛋白paxillin、p130Cas以及基质金属蛋白酶MMP-2与MMP-9蛋白的表达及活化情况。结果 FAK表达下调后,细胞黏附能力显著受到抑制,细胞迁移能力和侵袭能力均明显下降;细胞黏附分子p130Cas、paxillin的蛋白总量表达无明显改变,而磷酸化水平明显降低,其活化形式p-paxillin和p-p130Cas表达则明显受到抑制;MMP-2、MMP-9蛋白表达水平则在下调FAK表达后明显降低(P<0.01)。结论在人肝癌细胞HCC-LM3中,下调FAK表达可以影响肝癌细胞黏附迁移侵袭能力,其机制可能是通过调节相关细胞黏附分子的表达或活化来实现。

Objective To study the effect and mechanism of down-regulation of focal adhesion on cell adhesion, migration and invasion of hepatocellular carcinoma cells. Methods According to the different treatments, human HCC- LM3 cells were divided into untreated group（tumor cells were not treated）, control group（tumor cells were transfected with the empty vector） and FAK- sh RNA group（expression of FAK was stably low）. Cell adhesion, migration and invasion were tested by using cell adhesion assay, wound healing assay and Transwell assay, respectively. The expression and activation of cell adhesion and invasion related protein paxillin, p130 Cas, matrix metalloproteinase（MMP）-2 and MMP-9were detected by using Western blotting in HCC- LM3 cells（untreated group, control group and FAKsh RNA group）. Results Down-regulation of FAK could significantly reduce the ability of cell adhesion,migration and invasion. The activation of paxillin and p130 Cas were not influenced, while p-paxillin and p-p130 Cas were inhibited, and the expression of MMP-2 and MMP-9 were significantly decreased（P〈0.01）. Conclusion Down- regulation of FAK can affect cell adhesion, migration and invasion by regulating the expression or activation of cell adhesion molecules in human HCC-LM3 cells.