LIMK1高表达通过LIMK1/cofilin1通路促进人结肠癌SW480细胞增殖、迁移与侵袭

LIMK1 overexpression promotes the proliferation,migration and invasion through the LIMK1/cofilin1 pathway in human colon cancer SW480 cells

目的探究LIM激酶1(LIMK1)高表达通过LIMK1/cofilin1通路对人结肠癌SW480细胞生物学特性的影响。方法构建高表达LIMK1基因SW480细胞。实验分为空载体组、SW480组、LIMK1高表达组(LIMK1/SW480组)。RT-PCR、Western blotting分别检测LIMK1、Rac1、Pak1、p-LIMK1、cofilin1与p-cofilin1表达。MTT、流式细胞术、划痕实验和侵袭实验检测LIMK1高表达对SW480细胞增殖、细胞周期、迁移和侵袭能力的影响。结果与空载体组和SW480组比较,LIMK1/SW480组LIMK1 mRNA和蛋白表达、细胞增殖活性、穿膜细胞数升高(P<0.05),G2/M期细胞百分率、瘢痕距离减少(P<0.05)。LIMK1/SW480组LIMK1、p-LIMK1、p-cofilin1蛋白较空载体组和SW480细胞组升高(P<0.05)。各组Rac1、Pak1、cofilin1表达差异无统计学意义(P>0.05)。结论LIMK1高表达可能通过抑制LIMK1/cofilin1通路促进SW480细胞增殖、迁移、侵袭。

Aim To investigate the effect of LIMK1 overexpression on biological characteristics of human colon cancer SW480 cells through LIMK1/Cofilin1 pathway.Methods SW480 cells with highexpression of LIMK1 gene were constructed.The experiment was divided into empty vector group,SW480 group and LIMK1 high expression group(LIMK1/SW480 group).The expressions of LIMK1,Rac1,Pak1,p-LIMK1,CofilIn1 and p-CofilIn1 were detected by RT-PCR and Western blotting,respectively.MTT,flow cytometry,scratch assay and invasion assay were used to detect the effects of LIMK1 highexpression on proliferation,cell cycle,migration and invasion ability of SW480 cells.Results Compared with the empty vector group and SW480 group,the expression of LIMK1 mRNA and protein,cell proliferation activity and the number of transmembrane cells in LIMK1/SW480 group increased(P<0.05),while the percentage of G2/M cells and scar distance decreased(P<0.05).The expression of LIMK1,p-LIMK1 and p-cofilin1 in LIMK1/SW480 group were significantly higher than empty vector group and SW480 cell group(P<0.05).There was no significant difference in the expression of Rac1,Pak1 and cofilin1 in each group(P>0.05).Conclusion Overexpression of LIMK1 can promote the proliferation,migration and invasion of SW480 cells by inhibiting the LIMK1/Cofilin1 pathway.