去甲斑蝥素阻断αvβ6-ERK-ETS1信号通路抑制结肠癌细胞上皮-间质转化实验研究  

Norcantharidin inhibit colon cancer cell EMT process through blocking αvβ6-ERK-ETS1 signal pathway

作　　者：彭程[1] 牛正川 王奔[1] 高会杰[1] 牛卫博[1] 贺兆斌[1] 高超[1] 邹志进 马驰[1] 牛军[1] 

机构地区：[1]山东大学齐鲁医院肝胆外科,山东济南250012

出　　处：《中国现代普通外科进展》2014年第10期757-762,共6页Chinese Journal of Current Advances in General Surgery

基　　金：山东省优秀中青年科学家科研奖励基金项目(BS2011YY020)

摘　　要：目的:探讨去甲斑蝥素抑制结肠癌细胞上皮-间质转化的分子机制,为中药抗癌机制的研究奠定基础。方法:将HT-29结肠癌细胞经NCTD处理后,光镜下观察细胞形态学变化;流式细胞术检测处理前后细胞表面上皮表型及间质表型标志物的变化情况;Western Blotting分析常见调控细胞EMT过程信号传导通路中关键分子及其磷酸化状态的变化;转录因子活化技术观察核因子磷酸化状态的变化。结果:显微镜下观察发现,经NCTD处理后,结肠癌细胞EMT过程受到抑制;流式细胞术证实肿瘤细胞整合素αvβ6表达降低,同时上皮表型标志物表达升高,间质表型标志分子表达减少;Western Blotting分析常见调控细胞EMT过程信号传导通路中仅有ERK及p-ERK表达水平降低,其他关键蛋白无明显变化;转录因子活化技术发现利用si RNA技术干扰整合素αvβ6后,ERK下游的核转录因子中仅有ETS-1磷酸化水平改变,Western Blotting分析证实经NCTD处理后出现与干扰αvβ6表达造成的一致性改变。结论:去甲斑蝥素通过阻断αvβ6-ERK-ETS1信号通路抑制结肠癌细胞上皮-间质转化过程,从而发挥抗癌作用。Objective：To detect the molecular mechanism of NCTD inhibit colon cancer cell the EMT process,and make the foundation for researching other traditional Chinese medicine.Methods：Taking the HT-29 colon cancer cell with NCTD,observing the modification of cell morphology.Flow cytometry investigating the change of epithelium phenotype marker molecule and mesenchymal phenotype molecule.Western Blotting analysis the modification of crucial point protein and its phosphorylation which play an important role in regulating EMT process signal pathway.Transcription factor activation analysis observe the phosphorylation degree of nuclear factors.Results：After be treated with NCTD,the colon cancer cell EMT process was blocked.FACScan showed the the expression of integrin α v β 6 and mesenchymal phenotype molecule decreased in colon cancer cell.Otherwise,the expression of epithelium phenotype marker molecule increased.Western Blotting analysis showed only the expression of ERK and p-ERK changed in frequent signal pathway which regulating the EMT process.Transcription factor activation analysis showed that after decreased the expression of integrin αvβ 6 using siRNA technology,only the phosphorylation of ETS-1 changed,it＇s the downstream nuclear factor of ERK.Western Blotting analysis confirmed that NCTD can produce the same phenomena just like interfering with expression of integrin α v β 6.Conclusion：Norcantharidin can inhibit colon cancer cell EMT process through blocking α v β 6-ERK-ETS1 signal pathway,and so,fulfilling its anti-cancer drug mechanism.

关 键 词：去甲斑蝥素 整合素ΑVΒ6 转录因子ETS-1 结肠肿瘤 

分 类 号：R735.35[医药卫生—肿瘤] R730.5[医药卫生—临床医学]