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Apoptosis Mechanism of Human Cholangiocarcinoma Cells Induced by Bile Extract from Crocodile

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Abstract

Animal bile is popularly used as a traditional medicine in China, and bile acids are their major bioactive constituents. In the present study, effects of bile extract from crocodile gallbladder on QBC939 cell growth, cell cycle, and apoptosis were investigated by MTT assay, inverted microscopy, fluorescence microscopy, transmission electron microscopy, scanning electron microscopy, PI single- and FITC/PI double-staining flow cytometry, and western blotting. Our data have revealed that bile extract inhibited cells growth significantly, and the cell cycle was arrested in G1 phase. Bile extract induced QBC939 cell apoptosis, which was associated with collapse of the mitochondrial membrane potential and increase of ROS. In bile extract-treated cells, it was observed that the expression of bcl-2 decreased and cytochrome c released to cytosol, but the expression of bax remained unchanged. The data indicated that mitochondrial pathway might play an important role in bile extract-induced apoptosis in QBC939 cells. These results provide significant insight into the anticarcinogenic action of bile extract on cholangiocarcinoma cells.

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Abbreviations

CCA:

Cholangiocarcinoma

ROS:

Reactive oxygen species

ΔΨm:

Mitochondrial transmembrane potential

PI:

Propidium iodide

FBS:

Fetal bovine serum

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

HRP:

Horseradish peroxidase

ECL:

Enhanced chemiluminescence

TEM:

Transmission electron microscopy

SEM:

Scanning electron microscope

Rh123:

Rhodamine 123

DCFH-DA:

2,7-Dichlorofluorescein diacetate

DCF:

2,7-Dichlorofluorescein

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Acknowledgments

The present investigation was supported by Grant 81072014 of the Natural Science Foundation of China, National Foundation for fostering talents of basic science (J1030626) and supported by Sriracha Tiger Zoo Co., Ltd. Sriracha Thailand.

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Authors and Affiliations

  1. Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, School of Life Sciences, College of Environment and Ecology, Xiamen University, Xiamen, 361005, China

    Jin-He Kang, Wen-Qing Zhang, Wei Song, Dong-Yan Shen, Shan-Shan Li, Ling Tian, Yan Shi, Ge Liang & Qing-Xi Chen

  2. Sriracha Tiger Zoo Co., Ltd., P.O. Box 16, Sriracha, Chonburi, 20110, Thailand

    You-Xiong Xiong

Authors
  1. Jin-He Kang
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  2. Wen-Qing Zhang
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  3. Wei Song
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  5. Shan-Shan Li
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  7. Yan Shi
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  8. Ge Liang
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  9. You-Xiong Xiong
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  10. Qing-Xi Chen
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Correspondence to Qing-Xi Chen.

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Jin-He Kang and Wen-Qing Zhang contributed equally to this work.

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Kang, JH., Zhang, WQ., Song, W. et al. Apoptosis Mechanism of Human Cholangiocarcinoma Cells Induced by Bile Extract from Crocodile. Appl Biochem Biotechnol 166, 942–951 (2012). https://doi.org/10.1007/s12010-011-9482-x

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