Silencing expression of LIM kinase 1 inhibits invasion and migration of human glioma cells in vitro

Abstract

Objective To investigate the effect of silencing LIM kinase 1 (LIMK1) expresssion on invasion and migration of glioma cells, and investigate its possible mechanism. Methods Immunohistochemistry was used to detect the LIMK1 protein expression in normal brain tissues and glioblastoma tissues. Human glioma cells SNB19 were employed and assigned into three groups: siRNA-LIMK1 treated group (siRNA-LIMK1), siRNA-negative control group (siRNA-NC) and blank control group; siRNA-LIMK1 and non-sipencing siRNA were transfected into SNB19 cells by lipofectamine TM3000. The mRNA and protein expressions of LIMK1 were detected by real time-PCR and Western blotting, respectively. The migration and invasion of SNB19 glioma cells were evaluated by wound-healing assay and Transwell invasion assay. Immunofluorescence was used to localise the expression of LIMK1 in glioma cells and observe the lamellipodia morphology of glioma cells. The expressions of matrix metalloproteinase-2 (MMP-2), MMP-9, cofilin and phosphorylated-cofilin (p-cofilin) in SNB19 cells were determined by Western blotting. Results Immunohistochemistry showed that LIMK1 expressed in both normal brain tissues and glioblastoma tissues, but the LIMK1 expression was elevated in glioblastoma tissues as compared with that in normal brain tissues. The mRNA and protein expressions of LIMK1 in SNB19 cells of the siRNA-LIMK1 treated group were remarkably down-regulated as compared with those in the other two groups (P 0.05). Conclusions Reduction of LIMK1 expression effectively inhibits the invasion and migration abilities of SNB19 glioma cells in vitro, which suggests that LIMK1 might be a new therapeutic target for glioma. Key words: Glioma; Invasion; Migration; LIM kinase 1; RNA interference