二烯丙基二硫通过LIMK1-Cofilin1通路抑制人胃癌细胞侵袭与上皮-间质转化

Diallyl disulfide inhibits invasion and epithelial-mesenchymal transition in human gastric cancer cells through LIMK1-Cofilin1 pathway

目的:探讨二烯丙基二硫(DADS)对人胃癌MGC803细胞侵袭与上皮-间质转化(epithelial-mesenchymal transformation,EMT)的影响及其机制。方法:体内外实验分为MGC803细胞组与DADS处理组。MTT、划痕实验与侵袭实验检测DADS对增殖、迁移与侵袭的影响。Western Blot检测LIMK1-Cofilin1通路与EMT相关蛋白表达。相差显微镜观察MGC803细胞形态改变。裸鼠实验检测DADS对移植瘤的作用。结果:MTT显示,DADS可呈时间依赖性抑制MGC803细胞增殖(P<0.05)。划痕实验显示,DADS组细胞迁移距离(59.9±1.9)μm较MGC803组(127.1±2.0)μm明显缩短(P<0.01)。侵袭实验显示,DADS组穿膜细胞(30.0±2.6)个较MGC803组(48.3±2.5)个明显减少(P<0.01)。Western Blot显示,DADS作用6 h、12 h、24 h、48 h后,MGC803细胞LIMK1、p-LIMK1与p-Cofilin1(P<0.001),Vimentin(P<0.001)与MMP-9(P<0.001)分别呈时间依赖性下调,而E-cadherin(P=0.001)与TIMP-3明显上调(P<0.001)。相差显微镜显示,DADS处理后,纤维母细胞样细胞减少,异型性显著降低。裸鼠实验显示,DADS组移植瘤较MGC803细胞组生长缓慢(P<0.05),DADS组移植瘤重量较MGC803细胞组降低,抑瘤率为60.19%(P<0.001),Ki-67、Vimentin、Snail和CD34阳性表达均降低,E-cadherin表达增加。结论:DADS可通过LIMK1-Cofilin1通路体内外抑制MGC803细胞侵袭与EMT。

Objective:To investigate the effects of diallyl disulfide(DADS)on invasion and epithelial-mesenchymal transformation(EMT)of human gastric cancer MGC803 cells and its mechanism.Methods:In vitro and in vivo experiments were divided into MGC803 cell group and DADS treatment group.MTT,scratch test and invasion test were used to determine the effects of DADS on proliferation,migration and invasion.The expression of EMT-related proteins in LIMK1-Cofilin1 pathway was detected by Western Blot.The morphological changes of MGC803 cells were observed by phase contrast microscopy.The effects of DADS on tumor transplantation were tested in nude mice.Results:MTT showed that DADS could inhibit MGC803 cell proliferation in a time-dependent manner(P<0.05).Scratch test showed that the cell migration distance of DADS group was(59.9±1.9)μm shorter than that of MGC803 group(127.1±2.0)μm(P<0.01).Invasion test showed that DADS group had(30.0±2.6)transmembrane cells significantly decreased compared with MGC803 group(48.3±2.5)(P<0.01).Western Blot showed that after DADS treatment for 6 h,12 h,24 h and 48 h,LIMK1,p-LIMK1 and p-Cofilin1(P<0.001),Vimentin(P<0.001)and MMP-9(P<0.001)were down-regulated in a time-dependent manner,while E-cadherin(P=0.001)and TIMP-3 were significantly up-regulated(P<0.001)in MGC803 cells respectively.Contrast microscopy showed that after DADS treatment,the fibroblast-like cells decreased and the atypia decreased significantly.In nude mice,the tumor growth of DADS group was slower than MGC803 cell group( P <0.05),the tumor weight of DADS group was lower than MGC803 cell group,the tumor inhibition rate was 60.19%( P <0.001),and the positive expression of Ki-67,Vimentin,Snail and CD34 decreased,and the expression of E-cadherin was increased. Conclusion: DADS can inhibit MGC803 cell invasion and EMT through LIMK1-Cofilin1 pathway in vitro and in vivo.