肝细胞生长因子对肝细胞癌细胞系SMMC-7721黏着斑激酶的影响  

Effects of hepatocyte growth factor on the expression and phosphorylation of focal adhesion kinase in SMMC-7721 cells

作　　者：苏荣健[1] 李贞[2] 李宏丹[1] 宋慧娟[1] 程留芳[2] 

机构地区：[1]辽宁医学院科学实验中心辽宁省高校分子细胞生物学和新药开发重点实验室,辽宁省锦州市121001 [2]中国人民解放军总医院临床部消化科,北京市100853

出　　处：《世界华人消化杂志》2009年第20期2070-2073,共4页World Chinese Journal of Digestology

基　　金：辽宁省博士启动基金资助项目;No.20061074;辽宁省教育厅重点实验室基金资助项目;No.2008S142~~

摘　　要：目的:探讨肝细胞生长因子(HGF)对黏着斑激酶(FAK)、Src表达及活性的影响以及PI3K在该过程中的作用.方法:LY294002预处理阻断PI3K的活性、HGF处理SMMC-7721后检测FAK和Src的表达、磷酸化及在细胞内的分布.应用免疫印迹技术检测FAK、Src的表达及磷酸化,免疫荧光技术检测FAK在SMMC-7721细胞中的分布.结果:HGF(50μg/L)可以促进FAKY397位点的磷酸化,对FAK的表达没有影响.应用PI3K抑制物LY294002处理后,FAKY397的磷酸化水平显著降低.HGF处理后,FAK主要位于细胞边缘,呈簇状分布,应用PI3K抑制物,FAK在细胞内弥散分布.HGF处理后,Src激酶和SrcY416的磷酸化水平明显增加,而经LY294002处理后,Src激酶与SrcY416的磷酸化水平明显降低.结论:肝细胞癌中,HGF以PI3K依赖性的方式促进FAK和Src的活化.AIM： To investigate the effects of hepatocyte growth factor （HGF） on the expression and phosphorylation of focal adhesion kinase （FAK） and Src kinase, and explore the role of phosphatidylinositol 3-kinase （PI3K） in this process. METHODS： After hepatocellular carcinoma cells （SMMC-7721） were pretreated with LY294002 and stimulated with HGF, the phosphorylation status of FAK at Y397 and Src at Y416 were ana- lyzed by Western blot, and the distribution of FAK was observed by immunofluorescence.RESULTS： HGF （50 μg/L） was able to promote the phosphorylation of FAK at Y397, but had no effect on the expression of FAK. Pretreatment of cells with LY294002 （an inhibitor of PI3K） significantly decreased the phosphorylation of FAK at Y397. After HGF treatment, FAK was mainly clustered in the periphery of hepatocellular carcinoma cells. In contrast, FAK was diffused throughout the cells after pretreatment with PI3K inhibitor. HGF treatment could significantly raise the phosphorylation levels of Src kinase and Src-Y416 while pretreatment with LY294002 could significantly decrease the phosphorylation levels of Src kinase and Src-Y416. CONCLUSION： HGF is able to promote the activation of FAK and Src in a PI3K-dependent manner.

关 键 词：肝细胞癌 肝细胞生长因子 黏着斑激酶 PI3K 

分 类 号：R735.7[医药卫生—肿瘤]