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

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	Apoptosis of human cholangiocarcinoma cells induced by ESC-3 from Crocodylus siamensis bile
	Song, Wei ; Shen, Dong-Yan ; Kang, Jin-He ; Li, Shan-Shan ; Zhan, Hui-Wang ; Shi, Yan ; Xiong, You-Xiong ; Liang, Ge ; Chen, Qing-Xi ; Chen QX(陈清西)
刊名	http://dx.doi.org/10.3748/wjg.v18.i7.704
	2012-02-21
关键词	INTRAHEPATIC CHOLANGIOCARCINOMA CHOLANGIOCYTE GROWTH CANCER CELLS DNA-DAMAGE ACID RATS ACTIVATION INDUCTION SECRETION MORTALITY
英文摘要	National Natural Science Foundation of China [81072014]; National Foundation for Fostering Talents in Basic Sciences [J1030626]; Thailand Sriracha Tiger Zoo Co. Ltd., Sriracha, Thailand; 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 (V-FITC)/PI staining. Rh123 staining was used to detect the alteration of mitochondrial membrane potential (Delta psi m). 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 mu g/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. (C) 2012 Baishideng. All rights reserved.
语种	英语
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/65654]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	Song, Wei,Shen, Dong-Yan,Kang, Jin-He,et al. Apoptosis of human cholangiocarcinoma cells induced by ESC-3 from Crocodylus siamensis bile[J]. http://dx.doi.org/10.3748/wjg.v18.i7.704,2012.
APA	Song, Wei.,Shen, Dong-Yan.,Kang, Jin-He.,Li, Shan-Shan.,Zhan, Hui-Wang.,...&陈清西.(2012).Apoptosis of human cholangiocarcinoma cells induced by ESC-3 from Crocodylus siamensis bile.http://dx.doi.org/10.3748/wjg.v18.i7.704.
MLA	Song, Wei,et al."Apoptosis of human cholangiocarcinoma cells induced by ESC-3 from Crocodylus siamensis bile".http://dx.doi.org/10.3748/wjg.v18.i7.704 (2012).