【摘要】 目的 探讨二烯丙基二硫(DADS)抑制人结肠癌SW480细胞侵袭和上皮间质转化(EMT)的可能机制。方法 免疫组化检测结肠癌组织中尿激酶型纤溶酶原激活剂(uPA)受体(uPAR)的表达。基因转染技术构建uPAR过表达细胞。实验分为SW480组、SW480+DADS组、uPAR组、uPAR+DADS组。迁移、侵袭实检测各组细胞迁移与侵袭能力。RT-PCR、Western blotting分别检测相关信号通路分子mRAN及其蛋白的表达。裸鼠实验验证DADS对uPAR过表达SW480细胞移植瘤的影响。结果 结肠癌组织中uPAR表达显著高于癌旁正常黏膜组织(P<0.05)。成功构建稳定uPAR过表达SW480细胞。uPAR过表达上调uPA、uPAR表达,而DADS下调uPA、uPAR表达。uPAR过表达增强细胞迁移、侵袭能力,DADS抑制uPAR过表达细胞的迁移、侵袭能力。DADS通过下调Vimentin、基质金属蛋白酶(MMP-9)表达,上调E-cadherin、组织金属蛋白酶抑制物(TIMP)-3表达抑制SW480细胞EMT。裸鼠实验结果显示,DADS可体内抑制uPAR过表达SW480细胞EMT。结论 DADS通过负调控uPAR/uPA信号体内外抑制人结肠癌SW480细胞侵袭和EMT。更多还原

【Abstract】 Aim To explore the possible mechanism of diallyl disulfide(DADS) to inhibit invasion and epithelial-mesenchymal transition(EMT) of human colorectal cancer SW480 cells.Methods The expression of urokinase-type plasminogen activator(uPA) receptor(uPAR) in colon cancer tissues was detected by immunohistochemistry.uPAR overexpressing cells were constructed by gene transfection.The experiment was divided into SW480 group,SW480+DADS group,uPAR group,uPAR+DADS group.The cell migration and invasion ability in each group were detected through migration and invasion tests.The expression of mRAN and its protein in related signaling pathways was detected by RT-PCR and Western blotting.Nude mouse experiments were conducted to verify the effect of DADS on uPAR overexpressing SW480 cell transplantation tumors.Results The expression of uPAR in colon cancer tissues was significantly higher than that in adjacent normal tissues(P<0.05).SW480 cells with stable overexpression uPAR were constructed successfully.Overexpression of uPAR up-regulated the expression of uPA and uPAR,while DADS down-regulated the expression of uPA and uPAR.uPAR overexpression enhanced cell migration and invasion,whereas DADS inhibited the migration and invasion of uPAR-overexpression cells.DADS inhibited EMT in SW480 cells by down-regulating the expression of Vimentin and matrix metalloprotein(MMP)-9,and up-regulating the expression of E-cadherin and tissue inhibitor of metalloproteinase(TIMP)-3.The results of nude mouse experiments showed that DADS can inhibit uPAR overexpression in SW480 cells and EMT in vivo.Conclusion DADS inhibits human colon cancer SW480 cell invasion and EMT by negatively regulating uPAR/uPA signaling pathway in vivo and in vitro.更多还原