黏着斑激酶在Hep G2肝癌细胞黏附和迁移中的作用  

Role of Focal Adhesion Kinase in Adhesion and Migration of Hep G2 Cells

作　　者：马云龙[1] 沈阳[1] 任弘毅[1] 孙亨[1] 余泓池[1] 刘肖珩[1] 

机构地区：[1]四川大学华西基础医学与法医学院生物医学工程研究室,成都610041

出　　处：《生物医学工程学杂志》2013年第2期342-346,354,共6页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(10972148;11172198);国家基础科学人才培养基金能力提高资助项目(J1103604)

摘　　要：黏着斑激酶(FAK)在细胞的黏附和迁移中起着关键的作用,它通过调节下游小G蛋白实现对细胞黏附和迁移的调控。本研究采用FAK抑制剂阻断FAK在Y397位点的磷酸化;细胞损伤模型测定不同浓度(0～250nmol/mL)FAK抑制剂在0、2、4、8、24h各时间点对Hep G2细胞迁移的影响;免疫荧光结合Western blot测定50nmol/mL的FAK抑制剂处理Hep G2细胞120min后,细胞骨架的变化及小G蛋白Rac1、RhoA和Cdc42表达的影响。结果表明,FAK抑制剂对肝癌细胞迁移的抑制作用呈浓度和时间依赖性,抑制Hep G2细胞FAK磷酸化120min后,细胞骨架发生明显改变,而对小G蛋白的表达在不同时间段的抑制作用呈现出不同的时间效应,抑制FAK磷酸化可以下调下游小G蛋白的表达,从而抑制肝癌细胞的黏附和迁移行为。以上结果揭示FAK抑制剂通过阻止FAK的磷酸化抑制肿瘤细胞的迁移。该研究结果提示FAK抑制剂可能会抑制肿瘤细胞从原发灶向转移灶的迁移,从而在恶性肿瘤的预防和治疗中具有潜在的应用价值。Focal adhesion kinase （FAK） plays a critical role in the process of cell adhesion and migration by regulating the expression of downstream small G proteins. A kind of focal adhesion kinase （FAK） inhibitor was used to inhibit the phosphorylation of Y397 site of FAK, and scratch wound migration assay was used to examine the effect of FAK inhibitor with different concentrations （0-250 nmol/mL） on the migration of hepatomal cells （Hep G2 ceils） at 0, 2, 4, 8 and 24h. Immunofluorescenee analysis and Western blot analysis were performed to detect the expression of F- actin and small G proteins Roe1, RhoA and Cdc42 in Hep G2 cells treated with FAK inhibitor for 120min. The re- sults indicated that the FAK inhibitor can inhibit the migration of Hep G2 cells with a dose- and time-dependent man- ner, F-actin was down-regulated in Hep G2 cells treated with FAK inhibitor for 120mJn, and expression of small G proteins were inhibited at different durations. The inhibition of FAK phosphorylation could inhibit cell adhesion and migration by down-regulating small G proteins. These results suggested that FAK inhibitor can inhibit the migration of tumor cells by blocking FAK phosphorylation. This means that FAK inhibitor can block the metastasis of tumor cells to surrounding tissues. It may be a potential application in the prevention and treatment of cancer.

关 键 词：黏着斑激酶 小G蛋白 肝癌细胞 细胞黏附 细胞迁移 

分 类 号：R735.7[医药卫生—肿瘤]