朱剑梅,金 科.辣椒碱对结肠癌细胞自噬抑制作用的研究[J].肿瘤学杂志,2014,20(6):477-482.
辣椒碱对结肠癌细胞自噬抑制作用的研究
The Inhibition Effect of Capsaicin on Autophagy in Colon Cancer Cell
投稿时间:2013-09-17  
DOI:10.11735/j.issn.1671-170X.2014.06.B009
中文关键词:  结肠肿瘤  细胞自噬  辣椒碱  信号通路
英文关键词:colon neoplasms  autophagy  capsaicin  signaling pathways
基金项目:
作者单位
朱剑梅 四川省医学科学院·四川省人民医院 
金 科 四川省医学科学院·四川省人民医院 
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中文摘要:
      摘 要:[目的] 研究辣椒碱对人结肠癌细胞SW480自噬的抑制作用。[方法] 通过MTT法和克隆形成实验检测不同浓度辣椒碱处理的细胞增殖率,免疫荧光检测细胞自噬体的形成,Western Blot检测自噬标志物和信号通路蛋白的表达。通过蛋白组实验方法获取辣椒碱药物的相互作用蛋白,质谱技术进行蛋白鉴定及STRING数据库进行生物信息学分析。[结果] MTT结果表明100μmol/L浓度的辣椒碱能促进细胞增殖,该浓度的辣椒碱抑制了细胞自噬体的形成,抑制了 LC3 的活化,促进了p62的表达,提高了Akt和mTOR的磷酸化。质谱分析和STRING数据库查询发现了7个辣椒碱的相互作用蛋白,其中ATG2B与自噬存在重要联系。[结论]低浓度辣椒碱通过激活PI3K-Akt信号通路及与ATG2B相互作用来抑制细胞自噬。
英文摘要:
      Abstract:[Purpose] To investigate the inhibition effect of capsaicin on autophagy in human colon cancer cell line SW480. [Methods] MTT and colony formation assays were used to examine the effect of cell proliferation under treatment with various concentrations of capsaicin. The formation of autophagosomes was performed by immunofluorescent staining. Western Blot was conducted to measure the altered levels of autophagy-related proteins and signaling pathways. Furthermore,proteomic approach was used to investigate the intracellular proteins that can interact with capsaicin. These proteins were identified by mass spectrometry and the bioinformatic analysis was further performed by String. [Results] Capsaicin(100μmol/L) was capable to induce cell proliferation of SW480. Moreover,capsaicin at the concentration of 100μmol/L could inhibit the formation of autophagosomes,suppress the activation of LC3,enhance the levels of p62,and induce the phosphrylation of Akt and mTOR. Proteomics analysis identified 7 proteins that directly interact with capsaicin,among which ATG2B was significantly involved in autophagic event. [Conclusion] Capsaicin could inhibit autophagy by activating PI3K-Akt signaling pathway and binding to ATG2B.
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