DADS downregulates the Rac1-ROCK1/PAK1-LIMK1-ADF/cofilin signaling pathway, inhibiting cell migration and invasion.
DADS downregulates the Rac1-ROCK1/PAK1-LIMK1-ADF/cofilin signaling pathway, inhibiting cell migration and invasion.
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DOI：
10.3892/or.2012.2168
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发表时间：
2013-02
期刊：
影响因子：
4.2
通讯作者：
Yujuan Zhou;Jian Su;Ling Shi;Q. Liao;Q. Su
中科院分区：
文献类型：
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作者：
Yujuan Zhou;Jian Su;Ling Shi;Q. Liao;Q. Su
本研究旨在探讨二烯丙基二硫化物(DADS)下调LIM激酶-1(LIMK1)从而抑制人结直肠癌细胞迁移和侵袭的分子机制。采用RNA干扰技术建立稳定的LIMK1-miRNA/SW480细胞系。通过划痕愈合实验和Transwell迁移实验观察DADS和LIMK1 RNA干扰对SW480细胞迁移和侵袭的影响。采用RT-PCR和免疫印迹方法检测DADS对SW480细胞中Rho-Rho激酶(ROCK)1/p21-激活的激酶(PAK)-1-Lim激酶(LIMK)-1-肌动蛋白解聚因子(ADF)/cofilin通路信号分子的影响。DADS(45 mg/L)可显著降低SW480细胞的愈合和迁移速度，并显著抑制细胞的穿透能力(P<；0.05)。免疫组织化学和Western印迹分析结果显示，DADS显著下调LIMK1蛋白表达，抑制LIMK1蛋白磷酸化。RT-PCR和Western印迹分析结果表明，DADS抑制了Rac1、ROCK1、PAK1、LIMK1和destrin的mRNA和蛋白表达，并抑制了LIMK1和cofilin 1的蛋白磷酸化。结果表明，LIMK1的表达与SW480细胞的迁移和侵袭能力呈正相关。DADS下调rac1-ROCK1/PAK1-LIMK1-ADF/cofilin信号通路，抑制SW480细胞的迁移和侵袭。
The aim of this study was to explore the molecular mechanisms of the diallyl disulfide (DADS)-mediated downregulation of LIM kinase-1 (LIMK1) and the consequent inhibition of the migration and invasion of human colorectal cancer cells. RNA interference technology was used to establish stable LIMK1-miRNA/SW480 cell lines. The effects of DADS and LIMK1 RNA interference on the migration and invasion of SW480 cells were observed by scratch wound healing assay and Transwell migration assay. The effects of DADS on signaling molecules of the Rac1-Rho kinase (ROCK)1/p21-activated kinase (PAK)1-LIM kinase (LIMK)1-actin depolymerizing factor (ADF)/cofilin pathway in SW480 cells were examined by RT-PCR and western blot analysis. The healing and migration rate of the SW480 cells was significantly reduced and the cell penetrating ability was significantly suppressed (P<0.05) following treatment with DADS (45 mg/l). The immunohistochemistry and western blot analysis results showed that DADS significantly downregulated LIMK1 protein expression and suppressed LIMK1 protein phosphorylation. Furthermore, the RT-PCR and western blot analysis results revealed that DADS suppressed Rac1, ROCK1, PAK1, LIMK1 and destrin mRNA and protein expression, as well as the protein phosphorylation of LIMK1 and cofilin 1. The data demonstrate that LIMK1 expression positively correlates with the SW480 cell migration and invasion ability. DADS downregulates the Rac1-ROCK1/PAK1-LIMK1-ADF/cofilin signaling pathway, suppressing SW480 cell migration and invasion.