Apoptosis of human cholangiocarcinoma cells induced by ESC-3 from Crocodylus siamensis bile  

作　　者：Wei Song Dong-Yan Shen Jin-He Kang Shan-Shan Li Hui-Wang Zhan Yan Shi You-Xiong Xiong Ge Liang Qing-xi Chen 

机构地区：[1]State KeyLaboratory of Cellular Stress Biology, School of Life Sciences,Xiamen University, Xiamen 361005, Fujian Province, China [2]Thailand Sriracha Tiger Zoo Co. Ltd., Srira-cha, Thailand

出　　处：《World Journal of Gastroenterology》2012年第7期704-711,共8页世界胃肠病学杂志（英文版）

基　　金：Supported by The National Natural Science Foundation of China,No.81072014;the National Foundation for Fostering Talents in Basic Sciences,No.J1030626;the Thailand Sriracha Tiger Zoo Co.Ltd.,Sriracha,Thailand

摘　　要：AIM： To investigate the effects of ESC-3 isolated from crocodile bile on the growth and apoptosis induction of human cholangiocarcinoma cells.AIM:To investigate the effects of ESC-3 isolated from crocodile bile on the growth and apoptosis induction of human cholangiocarcinoma cells.METHODS:ESC-3 was isolated from crocodile bile by Sephadex LH-20 and RP-18 reversed-phase column.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was conducted to determine the effects of ESC-3 on the proliferation of human cholangiocarcinoma cell lines (QBC939,Sk-ChA-1 and MZ-ChA-1).Giemsa staining,Hoechst 33258 and acridine orange/ethidium bromide staining showed the morphological changes of Mz-ChA-1 cells exposed to ESC-3 at different concentrations.Flow cytometry with regular propidium iodide (PI) staining was performed to analyze the cell cycle distribution of Mz-ChA-1 cells and to assess apoptosis by annexin v-fluorescein isothiocyanate (VFITC)/PI staining.Rh123 staining was used to detect the alteration of mitochondrial membrane potential (ΔΨm).The protein levels of Bax,Bcl-2,Cdk2,cytochrome c and caspase-3 were further confirmed by Western blotting.RESULTS:ESC-3 significantly inhibited the growth of three human cholangiocarcinoma cell lines and arrested Mz-ChA-1 cell cycle at G0/G1 phase.Mz-ChA-1 cells showed typical apoptotic morphological changes after treated with ESC-3 (10 μg/mL) for 48 h.Cell death assay indicated that Mz-ChA-1 cells underwent apoptosis in a dose-dependent manner induced by ESC-3.In addition,ESC-3 treatment could downregulate the protein level of Bcl-2 and upregulate the Bax,leading to the increase in the ratio of Bax to Bcl-2 in Mz-ChA-1 cells.Meanwhile,cytochrome c was released from the mitochondria into the cytosol,which subsequently initiated the activation of caspase-3.All these events were associated with the collapse of the mitochondrial membrane potential.CONCLUSION:ESC-3,the active ingredient of crocodile bile,induced apoptosis in Mz-ChA-1 cells through the mitochondria-dependent pathway and may be a potential chemotherapeutic drug for the treatment of cholangiocarcinoma.

关 键 词：Crocody/us s/amensis bile Cholangiocarci-noma ANTIPROLIFERATION APOPTOSIS MITOCHONDRIA 

分 类 号：S852.33[农业科学—基础兽医学] Q279[农业科学—兽医学]