二烯丙基二硫下调LIMK1抑制人结肠癌SW480细胞迁移与侵袭

摘要

Aim To investigate the effects of DADS on expression of LIMK1 and the influences on migration and invasion in human colon carcinoma SW480 cells.And to elucidate the molecular mechanism of the inhibitive effect of Diallyl disulfide on migration and invasion in cancer cells.Methods The effects of DADS on expression of LIMK1 in SW480 cells was detected by Immunocytochemistry, RT-PCR and Western blot.Migration potentials were examined by Wound Healing assay.Invasion potentials were determined by transwell membrane assays.Results Compared with the negative control group,the expression of LIMK1 mRNA was inhibited significantly in SW480 cells after treated with DADS examined by RT-PCR.LIMK1 protein was overexpressed in human Colon cancer SW480 cell lines.detected by Immunocytochemistry.And its expression was remarkably downregulated after treatment with DADS.Moreover.Western bolt showed that DADS reduced expressions of LIMK1 protein in a time-dose dependent manner.Wound Healing assays showed that as the increase of concentration of DADS.the scratch healing distances were broadening.The results indicated that DADS can inhibit the capability of migration in SW480 cell lines in a dose-dependent manner.Transwell invasion assays revealed that with the increase of quantities of DADS.the numbers of invaded cells were obviously decreased.Its results exhibited that DADS can inhibit the capability of invasion in SW480 cell lines in a dosedependent manner.Conclusions After treatment with DADS , we can see significant inhibitory expression levels of LIMK1 , suppression of migration and invasion capability in SW480 cell lines.The potential molecule mechanisms may be that DADS can downregulate expression levels of LIMK1.%目的 研究二烯丙基二硫(diallyl disulfide,DADS)对人结肠癌SW480细胞LIMK1表达及迁移、侵袭能力的影响,阐明其抑制肿瘤细胞迁移和侵袭的分子机制.方法 RT-PCR、Western blot、免疫细胞化学检测DADS对SW480细胞LIMK1表达的作用;划痕愈合和侵袭实验检测DADS对SW480细胞迁移、侵袭能力的影响.结果 RT-PCR结果显示,与对照组相比,DADS处理SW480细胞后其LIMK1 mRNA表达明显下调.免疫细胞化学检测发现LIMK1蛋白在人结肠癌SW480细胞中高表达,DADS作用后LIMK1蛋白表达明显下调.Western blot检测显示,DADS呈时间浓度依赖性下调SW480细胞LIMK1蛋白表达.划痕愈合实验结果显示,随着DADS浓度增高,细胞划痕距离增宽,表明DADS呈浓度依赖性抑制SW480细胞迁移能力.Transwell侵袭实验结果表明,随着DADS浓度增高,细胞穿膜数逐渐减少,表明DADS呈浓度依赖性抑制SW480细胞侵袭能力.结论 DADS下调LIMK1表达可抑制人结肠癌SW480细胞迁移、侵袭,DADS抑制人结肠癌SW480细胞迁移、侵袭的分子机制可能与降低LIMK1蛋白表达水平有关.