二烯丙基二硫抑制人胃癌SGC7901细胞侵袭及相关机制

Diallyl disulfide inhibit migration and invasion in human gastric cancer SGC7901 cells and its mechanism

目的探讨二烯丙基二硫(diallyl disulfide,DADS)对人胃癌SGC7901细胞迁移和侵袭的影响及其分子机制。方法划痕愈合和侵袭实验检测DADS对SGC7901细胞迁移与侵袭能力的影响;RT-PCR、Western blot检测DADS作用SGC7901细胞后MMP-9和TIMP-3表达。结果划痕实验结果显示,15 mg.L-1DADS处理SGC7901细胞12 h、24 h后,较未处理组划痕距离明显增加(P<0.05),表明DADS可抑制SGC7901细胞迁移。Transwell实验结果显示,15 mg.L-1DADS处理SGC7901细胞12 h与24 h后,穿膜细胞数分别为(31.9±0.46)、(23.5±0.52)个,较未处理组的(51.6±0.68)、(41.3±0.72)个明显减少(P<0.05),表明DADS可抑制SGC7901细胞侵袭能力。RT-PCR结果显示,15 mg.L-1DADS处理SGC7901细胞12 h与24 h后,MMP-9 mRNA表达明显下调(P<0.05),而TIMP-3 mRNA表达明显上调(P<0.05)。Western blot结果显示,15 mg.L-1DADS处理SGC7901细胞12 h、24 h、48 h后,MMP-9蛋白表达明显降低(P<0.05),而TIMP-3蛋白表达明显增加(P<0.05)。结论 DADS可抑制人胃癌SGC7901细胞迁移与侵袭能力,其机制可能与下调MMP-9和上调TIMP-3有关。

Aim To investigate the effects of migration and invasion and its molecular mechanism in human gastric cancer SGC7901 cells induced by diallyl disulfide（DADS）.Methods The effects of DADS for migration and invasion were examined by Wound Healing assay and Transwell assay,and the expression of mRNA and protein of MMP-9 and TIMP-3 were determined by RT-PCR and Western blot in SGC7901 cells,respectively.Results Wound Healing assay discovered that the scuffing distance of SGC7901 cells exposed to 15 mg·L-1 DADS for 12 h and 24 h was significantly increased compared with no-treatment group,indicating DADS could inhibit migration in SGC7901 cells（P0.05）.Transwell assay showed that the number of SGC7901 cells migrated through the Matrigel treated by 15 mg·L-1 DADS for 12 h and 24 h was 31.9±0.46 and 23.5±0.52 respectively,obviously decreased compared with 51.6±0.68 and 41.3±0.72 of no-treatment group,respectively,indicating DADS could inhibit invasion in SGC7901 cells（P0.05）.RT-PCR showed that expression of MMP-9 mRNA was down-regulated and expression of TIMP-3 mRNA was up-regulated in SGC7901 cells treated by 15 mg·L-1 DADS for 2 h and 24 h,respectively.Western blot demonstrated that the expression of MMP-9 protein was increased and the expression of TIMP-3 was decreased in SGC7901 cells exposed to 15 mg·L-1 DADS for 12,24 and 48 h in a time-dependent manner,respectively.Conclusion DADS may inhibit the migration and invasion in human gastric cancer SGC7901 cells,which may be related to the down-regulation of MMP-9 and up-regulation of TIMP-3.