δ-榄香烯通过线粒体途径诱导人结肠癌细胞DLD-1凋亡的研究  

Cell apoptosis induced by δ-elemene in colorectal adenoarcinoma cells via mitochondrial-mediated pathway

作　　者：王茜莎[1] 杨威 董金华[3] 臧林泉[1] 金桂芳[1] 张德志[1] 

机构地区：[1]广东药学院药科学院,广东广州510006 [2]广州医药工业研究院药物非临床评价研究中心,广东广州510240 [3]沈阳药科大学制药工程学院,辽宁沈阳110016

出　　处：《中国药理学通报》2010年第5期646-652,共7页Chinese Pharmacological Bulletin

基　　金：辽宁省教育厅基金资助项目(No202043228)

摘　　要：目的探讨δ-榄香烯诱导人结肠癌细胞DLD-1凋亡的作用及其机制。方法采用MTT法考察了δ-榄香烯对肿瘤细胞增殖的影响;ELISA法定量检测细胞DNA片段化;Annexin-VFITC/PI流式细胞术检测细胞PS外翻的影响;用Westernblot分析药物对蛋白质表达的影响。结果δ-榄香烯明显抑制DLD-1细胞的增殖。DNA核小体的检测、细胞PS外翻证实δ-榄香烯的抗肿瘤作用是以诱导肿瘤细胞凋亡的方式产生的;Bax蛋白转入线粒体内,细胞色素C的释放以及凋亡因子AIF从线粒体释放入胞质,δ-榄香烯激活caspase信号通路,引起pro-caspase-3蛋白水解为caspase-3,进一步发生PARP底物的断裂。结论δ-榄香烯通过线粒体途径诱导DLD-1细胞发生凋亡。Aim To investigate the apoptotic effect and mechanism of δ-elemene on Colorectal Adenoarcinoma Cells DLD-1 in vitro.Methods Cell proliferation was measured by MTT method.Cell DNA fragmentation was quantitatively assessed by ELISA assay.The levels of detectable phosphatidylserine（PS）externalization were quantified using flow cytometry by Annexin V FITC kit.Gene expressions of Bax,cytochrome C,pro-caspase-3 and AIF were determined by Western blot analysis.Results δ-elemene inhibited the proliferation of DLD-1 cell significantly.The apoptosis of DLD-1 cells was further confirmed and quantified by DNA fragmentation ELISA and Annexin V（AnV）binding of externalized PS using flow cytometry.In the process ofthe induction of apoptotic cell death,Bax translocated into mitochondria,a release of cyto C and apoptosis inducing factor（AIF）from mitochondria into the cytosol occurred,δ-elemene activated the caspase-signaling pathway,leading to the proteolysis conversion of pro-caspase-3 to activate caspase-3,and the subsequent cleavage of the caspase substrate PARP.Conclusion DLD-1 cell death by δ-elemene is through a mitochondrial-mediated pathway.

关 键 词：δ-榄香烯 人结肠癌细胞DLD-1 凋亡 PS外翻 线粒体途径 PARP 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学]