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Apoptosis Induced by Aqueous Extracts of Crocodile Bile in Human Heptacarcinoma SMMC-7721

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Abstract

In the present study, effects of aqueous extracts from Crocodylus siamensis bile (AE-CB) on SMMC-7721 cell growth, cell cycle, and apoptosis were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, inverted microscopy, fluorescence microscopy, propidium iodide (PI) single- and fluorescein isothiocyanate (FITC)/PI double-staining flow cytometry, and western blotting. Our data have revealed that AE-CB significantly inhibited the growth of SMMC-7721 cell and arrested cell cycle at G0/G1 phase. SMMC-7721 cells showed typical apoptotic morphological changes after treated with AE-CB for 48 h. Cell death assay indicated that SMMC-7721 cells underwent apoptosis in a dose-dependent manner induced by AE-CB. In addition, AE-CB 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 SMMC-7721 cells. Meanwhile, it was observed that the expression of Survivin and c-Myc decreased, but the expression of P53 increased. All these events were associated with increase of reactive oxygen species. The data indicated that mitochondrial pathway might play an important role in bile extract-induced apoptosis in SMMC-7721 cells. These results provide significant insight into the anticarcinogenic action of bile extract on SMMC-7721 cells.

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Abbreviations

ROS:

Reactive oxygen species

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

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Acknowledgments

The present investigation was supported by grant from the Natural Science Foundation of China (81101502), and GuangZhou TuoLong Biological Technology Co., Ltd. (XDHT2012106A).

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Authors and Affiliations

  1. State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, 361005, China

    Wei Song, Shan-Shan Li, Ping-ping Qiu, Dong-Yan Shen, Ling Tian, Qiu-Yan Zhang & Qing-Xi Chen

  2. GuangZhou TuoLong Biological Technology Co., Ltd., GuangZhou, 510801, China

    Long-Xing Liao

Authors
  1. Wei Song
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  2. Shan-Shan Li
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  3. Ping-ping Qiu
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Correspondence to Qing-Xi Chen.

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Wei Song and Shan-Shan Li contributed equally to this work.

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Song, W., Li, SS., Qiu, Pp. et al. Apoptosis Induced by Aqueous Extracts of Crocodile Bile in Human Heptacarcinoma SMMC-7721. Appl Biochem Biotechnol 170, 15–24 (2013). https://doi.org/10.1007/s12010-013-0120-7

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  • DOI: https://doi.org/10.1007/s12010-013-0120-7

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