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Effects of Helicobacter pylori γ-Glutamyltranspeptidase on Apoptosis and Inflammation in Human Biliary Cells

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Abstract

Background

Several studies have reported the presence of H. pylori in individuals with hepatobiliary diseases, but in vitro and in vivo studies are still needed. Here, we determined the effects of H. pylori γ-glutamyltranspeptidase (GGT) on the induction of apoptosis and IL-8 production in a human cholangiocarcinoma cell line (KKU-100 cells).

Methods

Cell viability and DNA synthesis were examined by MTT and BrdU assays, respectively. RT-PCR and western blot analysis were performed to assess gene and protein expression, respectively. IL-8 secretion in KKU-100 cells was measured by ELISA.

Results

Exposure to the H. pylori ggt + strain decreased KKU-100 cell survival and DNA synthesis when compared with cells exposed to the H. pylori ggt mutant strain. Treatment with recombinant H. pylori GGT (rHP-GGT) dramatically decreased cell survival and DNA synthesis, and stimulated apoptosis; these features corresponded to an increased level of iNOS gene expression in KKU-100 cells treated with rHP-GGT. RT-PCR and western blot analyses revealed that rHP-GGT treatment enhanced the expression of pro-apoptotic molecules (Bax, Caspase-9, and Caspase-3) and down-regulated the expression of anti-apoptotic molecules (Bcl-2 and Bcl-xL). The extrinsic-mediated apoptosis molecules, including Fas and activated Caspase-8, were not expressed after treatment with rHP-GGT. Furthermore, rHP-GGT significantly stimulated IL-8 secretion in KKU-100 cells.

Conclusion

Our data indicate that H. pylori GGT might be involved in the development of cancer in hepatobiliary cells by altering cell kinetics and promoting inflammation.

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Acknowledgments

We would like to thank the Commission on Higher Education, Thailand, for supporting this research with a grant funds under the Strategic Scholarships for Frontier Research Network for the Joint Ph.D. Program Thai Doctoral degree. We would also like to thank Khon Kaen University and the National R&D Program for Cancer Control, Ministry for Health, Welfare and Family affairs, Republic of Korea (0820050) for supporting some parts of this work. We would further like to thank the Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University for providing the cell line.

Conflict of interest

We declare that we have no conflicts of interest.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Wongwarut Boonyanugomol, Chariya Chomvarin & Chariya Hahnvajanawong

  2. Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen, 40002, Thailand

    Wongwarut Boonyanugomol, Chariya Chomvarin, Banchob Sripa & Chariya Hahnvajanawong

  3. Department of Microbiology, Gyeongsang National University School of Medicine, Chiram-dong 90, Jinju, South Gyeongsang, 660-751, Republic of Korea

    Jea-Young Song, Kyung-Mi Kim, Jung-Min Kim, Myung-Je Cho, Woo-Kon Lee, Hyung-Lyun Kang, Kwang-Ho Rhee & Seung-Chul Baik

  4. Research Institute of Life Science, Gyeongsang National University School of Medicine, Chiram-dong 90, Jinju, South Gyeongsang, 660-751, Republic of Korea

    Jea-Young Song, Kyung-Mi Kim, Jung-Min Kim, Myung-Je Cho, Woo-Kon Lee, Hyung-Lyun Kang, Kwang-Ho Rhee & Seung-Chul Baik

  5. Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Banchob Sripa

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  1. Wongwarut Boonyanugomol
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Correspondence to Seung-Chul Baik.

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Boonyanugomol, W., Chomvarin, C., Song, JY. et al. Effects of Helicobacter pylori γ-Glutamyltranspeptidase on Apoptosis and Inflammation in Human Biliary Cells. Dig Dis Sci 57, 2615–2624 (2012). https://doi.org/10.1007/s10620-012-2216-2

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  • DOI: https://doi.org/10.1007/s10620-012-2216-2

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