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Induction of cancer-related microRNA expression profiling using excretory-secretory products of Clonorchis sinensis

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Abstract

Clonorchis sinensis is a carcinogenic human liver fluke by which chronic infection is strongly associated with the development of cholangiocarcinoma. Although this cholangiocarcinoma is caused by both physical and chemical irritation from direct contact with adult worms and their excretory–secretory products (ESPs), the precise molecular events of the host–pathogen interactions remain to be elucidated. To better understand the effect of C. sinensis infection on cholangiocarcinogenesis, we profiled the kinetics of changes in cancer-related microRNAs (miRNAs) in human cholangiocarcinoma cells (HuCCT1) treated with C. sinensis ESPs for different periods. Using miRNA microarray chips containing 135 cancer-related miRNAs, we identified 16 miRNAs showing differentially altered expression following ESP exposure. Of these miRNAs, 13 were upregulated and 3 were downregulated in a time-dependent manner compared with untreated controls. Functional clustering of these dysregulated miRNAs revealed involvement in cell proliferation, inflammation, oncogene activation/suppression, migration/invasion/metastasis, and DNA methylation. In particular, decreased expression of let-7i, a tumor suppressor miRNA, was found to be associated with the ESP-induced upregulation of TLR4 mRNA and protein, which contribute to host immune responses against liver fluke infection. Further real-time quantitative PCR analysis using ESP-treated normal cholangiocytes (H69) revealed that the expressions of nine miRNAs (miR-16-2, miR-93, miR-95, miR-153, miR-195, miR-199-3P, let7a, let7i, and miR-124a) were similarly regulated, indicating that the cell proliferation and inhibition of tumor suppression mediated by these miRNAs is common to both cancerous and non-cancerous cells. These findings constitute further our understanding of the multiple cholangiocarcinogenic pathways triggered by liver fluke infection.

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Acknowledgments

We thank Drs. Dae Ghon Kim for donating H69 cells and Hye Ryung Jun for assistance in the early phases of the study. This study was supported by a National Research Foundation of Korea (NRF) grant funded by the Korea Government (MEST) (No. 2012R1A2A2A01014237).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Asan Institute for Life Sciences, University of Ulsan College of Medicine Asan Medical Center, 388-1 Punganp-2 dong, Songpa-gu, Seoul, 138-736, Republic of Korea

    Jhang Ho Pak, In Ki Kim, Seon Min Kim, Sejung Maeng & Kyoung Ju Song

  2. Department of Parasitology and Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju, 660-751, Republic of Korea

    Byoung-Kuk Na

  3. Department of Parasitology, Inha University School of Medicine, Incheon, 400-103, Republic of Korea

    Tong-Soo Kim

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  1. Jhang Ho Pak
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Correspondence to Jhang Ho Pak.

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Pak, J.H., Kim, I.K., Kim, S.M. et al. Induction of cancer-related microRNA expression profiling using excretory-secretory products of Clonorchis sinensis . Parasitol Res 113, 4447–4455 (2014). https://doi.org/10.1007/s00436-014-4127-y

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  • DOI: https://doi.org/10.1007/s00436-014-4127-y

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