辣椒碱通过上调NLRP3炎性小体诱导胃癌MGC-803细胞转移

Promotion effects of capsaicin on the human MGC-803 gastric cancer cells metastasis through upregulating the inflammatory corpuscle NLRP3

关键词:  辣椒碱  人胃癌细胞  NLRP3炎性小体  迁移

Key words:  capsaicin  human gastric cancer cells  inflammatory corpuscle NLRP3  metastasis

摘要:

[目的]探讨辣椒碱对胃癌MGC-803细胞转移的诱导作用及其对核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）炎性小体活化的影响。[方法]采用酶联免疫吸附（ELISA）法检测MGC-803细胞培养液中炎性因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）及肿瘤坏死因子-α（TNF-α）水平，蛋白免疫印迹（Western Blot）方法分析细胞中NLRP3炎性小体、细胞钙依粘连蛋白（E-cadherin）及波形蛋白（Vimentin）表达水平，采用Transwell法检测MGC-803细胞的迁移能力，流式细胞仪检测细胞内活性氧（ROS）含量。[结果]辣椒碱处理MGC-803后，培养液中炎性因子IL-1β、IL-6及TNF-α释放量明显升高、NLRP3表达量增加，同时间质样标志物Vimentin的表达升高、上皮样标志物E-cadherin表达降低，但用NLRP3特异性抑制剂MCC950处理细胞之后，炎性因子表达量下调、NLRP3及Vimentin蛋白表达减低、E-cadherin表达升高，并且MCC950处理后能抑制辣椒碱增加的细胞迁移数量；ROS抑制剂N-乙酰半胱氨酸（NAC）能逆转辣椒碱诱导结肠癌细胞NLRP3的蛋白水平及炎性因子释放。[结论]辣椒碱通过促进MGC-803细胞内ROS的表达水平来激活NLRP3炎性小体，进而促进胃癌细胞的迁移。

Abstract:

[Objective] To study the inducible effects of capsaicin on the human MGC-803 gastric cancer cells metastasis and the expression of the inflammatory corpuscle NLRP3.[Methods] The productions of inflammation-related molecules IL-1β,IL-6 and TNF-α in MGC-803 cells were detected by the enzyme-linked immunosorbent (ELISA) assay. The expression levels of the NLRP3,E-cadherin and Vimentin proteins in the cells were determined by Western Blot. The changes in the migration ability of MGC-803 cells were assessed by Transwell. The changes of ROS were detected by Flow Cytometer in MGC-803 cells.[Results] After the MGC-803 cells were treated with capsaicin,the productions of inflammatory cytokines and NLRP3 expression were greatly increased,and the mesenchymal marker Vimentin was also up-regulated,while the epithelial cell marker E-cadherin was down-regulated. Nevertheless,the expression of inflammatory cytokines,NLRP3 and Vimentin were inhibited by the NLRP3 specific inhibitor MCC950,which also inhibited the increase of the cell migration count induced by capsaicin. The ROS inhibitor NAC reversed the capsaicin-induced protein level of NLRP3 and inflammatory factor levels in MGC-803 cells.[Conclusion] Capsaicin promotes MGC-803 cells metastasis by increasing the expression of ROS in the gastric cancer cells to activate the inflammatory corpuscle NLRP3.