二烯丙基二硫上调RORα拮抗Wnt/β-catenin通路抑制人胃癌细胞EMT与迁移侵袭

Our previous studies demonstrated that diallyl disulfide (DADS) inhibited proliferation and migration and invasion of human gastric cancer cells by multiple signaling pathway. and 24 differential expression proteins were in gastric cancer MGC803 cells induced by DADS using proteome technique, and found that RORα is upregulated by DADS. RORα may be regulate the Wnt/β-catenin signaling by depending on PKCα signaling pathway. Wnt/β-catenin signaling is involved in tumor EMT and migration and invaision. On the basis the our previous researches, combined with the profissional knowledge, we hypothsis that DADS inhibits EMT and migration and invasion in human gastric cancer cells by upregulation of ROR, PKCα-dependent phosphorylation of ROR, negative regulation of TGF-β and Rac1 to resist Wnt/β-catenin pathway. These project will provide the theoretical basis for elucidating the mechanism of effect of DADS and development of the traditional Chinese medicine against gastric cancer and targeted intervention therapy.

我们前期研究表明，二烯丙基二硫（diallyl disulfide，DADS）可通过多种途径抑制胃癌细胞增殖与迁移侵袭，并运用蛋白质组学技术，获得胃癌细胞中可受DADS调控的24个差异蛋白质，其中，DADS可上调RORα表达。研究表明，RORα可能通过PKCα依赖的磷酸化调控经典的Wnt/β-catenin通路，而Wnt/β-catenin通路参与肿瘤细胞的EMT和侵袭转移。基于我们前期的工作基础，辅以大量的文献阅读，我们提出"DADS通过上调RORα，激活PKCα磷酸化RORα，负调控TGF-β和Rac1拮抗Wnt/β-catenin通路，抑制人胃癌细胞EMT与侵袭转移，RORα可能是DADS的作用靶点"的科学假设。通过该假设的验证，为进一步阐明DADS抑制胃癌迁移侵袭的机制，开发中药抗胃癌药物和靶向干预治疗提供理论基础。

本研究探讨二烯丙基二硫（DADS）上调RORα拮抗Wnt/β-catenin通路抑制人胃癌细胞EMT与迁移侵袭。结果显示，DADS可上调RORα，DADS 与RORα高表达可抑制MGC803细胞增殖与迁移侵袭，下调Wnt1，增加RORα与β-catenin结合，降低核内β-catenin与p-β-catenin表达，下调TCF-4及Axin、c-Myc、c-Jun、MMP-9，上调TIMP3，抑制c-Myc启动子活性，下调Snail与Vimentin和上调E-cadherin，细胞呈圆形或椭圆形，抑制裸鼠移植瘤生长，下调Ki-67、Vimentin、CD34，上调E-cadherin。证实DADS上调RORα可拮抗Wnt/β-catenin通路抑制人胃癌细胞EMT与侵袭。.DADS与TPA处理组RORα，p-RORα，PKCα，p-PKCα上调。DADS及TPA能促进PKCα与RORα的结合，下调Axin、c-Jun、Cyclin1、c-Myc，GO6976作用相反。表明DADS可通过PKCα/RORα介导抑制Wnt/β-catenin通路。 .DADS、重组人TGF-β蛋白、SB-431542与NSC23766处理MGC803细胞，显示DADS可下调TGF-β1、Rac1、β-catenin、Vimentin和上调E-cadherin，抑制EMT，移植瘤生长减慢，E-cadherin增加，Vimentin、Ki-67与CD34降低。证明DADS可下调TGF-β与Rac1通过Wnt/β-catenin通路抑制人胃癌细胞EMT。.DADS与SR1078能上调RORα表达，诱导细胞变圆，增殖与迁移侵袭能力下降，下调Rac1、TGF-β1、Vimentin和上调E-cadherin，RORα与β-catenin结合增加，核内β-catenin降低，证明DADS具有RORα激动剂的作用。