苦马豆素诱导人喉癌Hep-2细胞凋亡研究  

Effect of Swainsonine on Apoptosis of Human Laryngeal Cell Line Hep-2

作　　者：李莎[1] 张思楠[1] 杨军[1] 伍春莲[1,2] LI Sha ZHANG Si-nan YANG Jun WU Chun-lian(Key Laboratory of Southwest China Wildlife Resources Conservation （Ministry of Education）, China West Normal University,Nanchong 637009, China State Key Laboratory Breeding Base of Eeo-Environments and Bio-Resourees of the Three Gorges Reservoir Region,Southwest University,Chongqing 400715 ,China)

机构地区：[1]西华师范大学西南野生动植物资源保护省部共建教育部重点实验室,南充637009 [2]西南大学三峡库区生态环境与生物资源省部共建国家重点实验室,重庆400715

出　　处：《天然产物研究与开发》2016年第9期1384-1389,1383,共7页Natural Product Research and Development

基　　金：中国博士后基金(2013M540391);四川省教育厅重大培育项目(13CZ0029);三峡库区生态环境与生物资源省部共建重点实验室开放课题(SKL-2011-05)

摘　　要：苦马豆素具有广泛的抗肿瘤作用和免疫调节作用。本研究对苦马豆素诱导人喉癌Hep-2细胞凋亡的作用及其机制进行研究。通过不同浓度的苦马豆素处理Hep-2细胞24 h后,采用MTT法检测细胞活性;荧光显微镜观察细胞形态;Annexin V-FITC/PI双标记检测细胞凋亡;JC-1检测线粒体膜电位;实时荧光定量PCR和Western blot分别检测Bax和XIAP基因mRNA和蛋白质表达水平。在一定浓度范围内,苦马豆素抑制Hep-2细胞增殖,且呈剂量依赖性;荧光显微镜下细胞发生形态改变,并呈现典型的凋亡细胞特征;磷脂酰丝氨酸外翻加剧;线粒体膜电位逐渐下降;Bax的mRNA和蛋白质表达水平上升,XIAP的mRNA和蛋白质表达水平下降。苦马豆素通过调节细胞内Bax和XIAP的表达,并经线粒体信号途径诱导Hep-2细胞凋亡。Swainsonine have a wide range of antitumor and immunomodulatory effects. The apoptosis-inducing effect of swainsonine on human laryngeal carcinoma Hep-2 cells and its mechanisms were investigated in this study. Different concentrations of swainsonine was applied to Hep-2 cells for 24 h,the cells activity was detected by MTT assay. Cells morphology was observed under a fluorescence microscope. Cell apoptosis was detected by Annexin V-FITC / PI double labeling. JC-1 was used to detect the mitochondrial membrane potential. The mRNA and protein levels of Bax and XIAP were measured by Real-time PCR and Western blot respectively. Swainsonine inhibited the proliferation and induced the apoptosis of Hep-2 cells in a dose-dependent manner in a certain concentration range. The different levels of morphology change of apoptosis were observed. The eversion of phosphatidyl serine was aggravatingly intensified,and mitochondrial membrane potential decreased. The mRNA and protein expression level of Bax increased. The mRNA and protein expression level of XIAP decreased. Hence,it was concluded that swainsonine induced apoptosis through the mitochondrial pathway and dependent on the regulation of the proteins levels of Bax and XIAP.

关 键 词：苦马豆素 HEP-2细胞 细胞凋亡 线粒体途径 

分 类 号：Q291[生物学—细胞生物学]