FRK通过FAK信号通路抑制脑胶质瘤细胞迁移与侵袭作用的研究

目的研究酪氨酸相关激酶(fyn-related kinase,FRK)是否通过粘着斑激酶(focal adhesion kinase,FAK)信号通路抑制脑胶质瘤细胞的迁移和侵袭。方法采用Western blot(WB)检测FRK质粒的转染效果,细胞划痕和Transwell侵袭实验检测过表达FRK及下调FAK对胶质瘤U251细胞迁移和侵袭能力的影响;WB检测过表达FRK对FAK、P-FAK蛋白水平的影响以及下调FAK对FRK蛋白水平的影响。结果FRK质粒转染成功。过表达FRK后胶质瘤U251细胞的迁移与侵袭能力下降。FAK siRNA干扰效率达到70%,下调FAK后胶质瘤U251细胞的迁移与侵袭能力下降。过表达FRK后P-FAK的蛋白水平明显下降,而FAK蛋白水平无明显变化,下调FAK对FRK的蛋白水平无明显影响。结论 FRK可能通过抑制FAK的磷酸化来调控胶质瘤的侵袭与迁移。

FRK inhibits human glioma cells invasion and migration through FAK regulation

Objective To explore the mechanism of the inhibitory effect of fyn-related kinase ( FRK) on the glioma cells migration and invasion through focal adhesion kinase ( FAK) signaling pathways .Methods Western blot was used to examine the transfection efficiency of FRK plasmid . The effect of FRK over-expression or FAK down-regulation on the glioma U251 cells migration and invasion ability was examined by wound healing assay and transwell invasion assay respectively .The effect of FRK over-expression on the protein level of FAK , P-FAK, and the influence of FAK down-regulation on FRK protein level were analyzed by WB .Results The FRK plasmid was transfected into glioma U251 cells successfully , and FRK over-expression decreased the glioma cells migration and invasion .The efficiency of FAK reached to about 70%, and FAK down-regulation decreased the migratory and invasive ability of glioma U 251 cells.FRK over-expression decreased the protein level of P-FAK significantly , but had no effect on the total protein levels of FAK .In addition, the protein levels of FRK had no change after FAK down-regulation .Conclusion FRK may regulate the migration and invasion of the glioma cells by dephosphorylating the focal adhesion kinase .