整合蛋白α5,β1亚基的过表达对肝癌细胞粘附和迁移的影响

Effect of integrin α5,β1 gene over-expression on cell adhesion and migration in hepatocellular carcinoma cell line

目的 :研究整合蛋白α5 ,β1亚基的过表达与肝癌细胞粘附能力、迁移能力、集落形成能力、以及与肝癌细胞凋亡的关系。方法 :用已建立的单独转染α5 ,β1整合蛋白亚基的细胞株α5 3 772 1,β1 6 772 1和共转α5和 β1整合蛋白亚基的细胞株α5 β1 6 772 1;采用结晶紫染色法测定细胞的粘附能力、细胞的迁移能力 ;应用荧光染色法和流式细胞仪法测定细胞凋亡速度。结果 :转染后细胞与主要胞外基质蛋白 纤连蛋白的粘附分别为对照细胞的 1.39,1.34,1.7倍 ;与层粘连蛋白 (Ln)的粘附为对细胞的 1.3、2 .6、1.98倍 ;α5 ,β1整合蛋白转染株细胞在Fn上的迁移能力分别增加 1.2 2、0 .78、1.38倍。集落形成能力分别下降 38% ,35 %和 5 2 %。在无血清培养时 ,α5 ,β1整合蛋白转染株细胞的凋亡百分率分别提高 2 .4、0 .5 3、1.79倍。结论 :将α5 β1整合蛋白导入人肝癌细胞引起α5 β1整合蛋白过表达 ,可显著改变细胞的生长 ,粘附和迁移能力 ,从而导致体外集落形成能力明显下降 。

Purpose:To study the correlation between the over expression of α5,β1 integrin subunit and the ability of liver cancer cells for adhesion, migration, colony forming, and carcinoma cell apoptosis. Methods:Using the established α5/3 7721,β1/6 7721 cell strains transfected with α5 or β1 integrin subunit separately and α5,β1 subunits at the same time, we determined the ability of adhesion migration by crystal purple dying and the speed of cell apoptosis flow cytometry and fluorescent staining. Results:Adhesion to fibronectin of transfectants were 1.39,1.34,1.7 fold of control cells, while adhesion to laminin were stronger by 1.3,2.6,1.98 fold as compared with control cells. Migration to fibronectin increased 1.22,0.78,1.36 fold as compared with control cells. Colony forming ability decreased 38%, 35% and 52%. When cells were grown in serum free medium, the percentage of apoptosis cells in over express α5 and β1 integrin of hepatocellular carcinoma cells increased 2.4,0.53 and 1.79 fold separately . Conclusions:These results indicate that over expression of α5 β1 integrin may obviously change proliferation, adhesion, and migration of 7221 cells, and result in decrease of colony forming ability. It shows that α5,β1 integrin is one of the important molecules affecting malignant phenotypes of cells.