摘要
目的研究二烯丙基二硫(diallyl disulfide,DADS)对人胃癌MGC803细胞迁移侵袭及LIMK1表达的影响。方法MTT、划痕愈合和侵袭实验分别检测DADS对MGC803细胞增殖、迁移与侵袭能力的作用;RT-PCR、Western blot与免疫细胞化学检测LIMK1表达。结果 MTT显示,30 mg.L-1DADS作用MGC803细胞24、48、72、96 h后,增殖抑制率分别为31.8%、66.1%、83.6%、89.2%,呈明显的时间依赖关系(P<0.05)。划痕实验显示,10、20、30、40、50 mg.L-1DADS分别作用MGC803细胞,划痕愈合明显慢于对照组,呈剂量依赖关系(P<0.05)。侵袭实验显示,10、20、30、40、50 mg.L-1DADS作用MGC803细胞24 h后,穿过基质胶的细胞数分别为(35.8±3.74)、(34.1±2.02)、(31.7±4.81)、(17.2±3.08)、(13.2±3.36)个,较对照组(39.5±2.99)个数明显减少,呈剂量依赖性(P<0.05)。RT-PCR显示,30 mg.L-1DADS与MGC803细胞作用48 h后,LIMIK1mRNA明显下调(P<0.05)。Western blot与免疫细胞化学检测显示,30 mg.L-1DADS作用MGC803细胞6、12、24、48h后,LIMK1蛋白的表达水平呈时间依赖性下降(P<0.05)。结论 DADS可抑制人胃癌MGC803细胞迁移与侵袭能力,其机制可能与下调LIMK1有关。
Aim To investigate the effects of DADS on the migration and invasion of human gastric cancer MGC803 cells and the effects on LIMK1 expression.Methods Abilities of proliferation,migration and invasion in MGC803 cells by DADS were examined by MTT test,Wound healing assay and transwell membrane assays.The effects of LIMK1 expression in MGC803 cells by DADS were detected by RT-PCR and Western blot.Results MTT test showed that the inhibitory rates were 31.8%,66.1%,83.6% and 89.2%,respectively for 24 h,48 h,72 h and 96 h in MGC803 cells exposed to 30 mg·L-1 DADS,which exhibited a time-dependent model（P0.05）.Wound healing assays displayed that the scratch healing distances were significantly broadening in a dose-dependent manner,compared with the control group for 24 h in MGC803 cells treated with 10,20,30,40,50 mg·L-1 DADS（P0.05） respectively.Transwell experiments exhibited that the numbers of treated group cells through matrigel were 35.8±3.74,34.1±2.02,31.7±4.81,17.2±3.08 and 13.2±3.36,respectively,significantly lower than those in the control group（39.5±2.99） in a dose-dependent manner（P0.05）.Compared with the negative control group,the expression of LIMK1 mRNA was significantly inhibited in MGC803 cells after treated with 30 mg·L-1 DADS examined by RT-PCR（P0.05）.Moreover,Western blot showed that DADS reduced expressions of LIMK1 protein in a time-dependent manner（P0.05）.Conclusions DADS can significantly inhibit the capability of migration and invasion of MGC803 cells.The potential molecular mechanisms may be related to the downregulation of LIMK1 expression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2012年第12期1714-1718,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81102854)
湖南省教育厅科学研究重点项目(No 09A077)
