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Mucin-producing hamster cholangiocarcinoma cell line, Ham-2, possesses the aggressive cancer phenotypes with liver and lung metastases

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Abstract

Cholangiocarcinoma (CCA) is an aggressive bile duct cancer. Opisthorchis viverrini (O. viverrini) infection is a significant cause of CCA in the Greater Mekong subregion. Currently, there is no standard chemotherapeutic regimen for CCA. A unique hamster carcinogenesis model of O. viverrini–associated CCA was established. Molecular targets identified from the hamster CCA-comparative model are valuable for target identification and validation. Hamster CCA was induced by the administration of O. viverrini metacercariae and N-nitrosodimethylamine. Hamster-derived cancer cells were isolated and continuously cultured for more than 6 months. Ham-2 cell line was established and characterized in vitro and in vivo. Ham-2 exhibited chromosome hyperploidy. A comparative study with previously established cell line, Ham-1, demonstrated that Ham-2 acquired slower growth, higher adhesion, higher migration, and resistance to doxorubicin and 5-fluorouracil (5-FU). In BALB/c Rag-2/Jak3 double-deficient (BRJ) mice, Ham-2 subcutaneous transplantation formed mucin-producing cancers, which morphologically resemble human tubular cholangiocarcinoma. Intravenous-injected Ham-2 established the metastatic nodules in the lungs and livers of BRJ mice. Altogether, a new hamster cholangiocarcinoma cell line, Ham-2, which acquired more aggressive phenotypes in vitro and in vivo, was established. This cell line might be a valuable tool for comparative drug target identification and validation.

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Funding

This work was supported in part by e-ASIA Joint Research Program from Japan Agency for Medical Research and Development (AMED), Japan (Grant Number: 20jm0210062h0003 to SO) and from the National Science and Technology Development Agency, Thailand (Grant Number: FDA-CO-2562–9172-TH to KV). KV was partly supported by the Thailand Research Fund (Grant Number: RSA6180068).

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Authors and Affiliations

  1. Division of Hematopoiesis, Graduate School of Medicine and Joint Research Center for Human Retrovirus Infection, Kumamoto University, Kumamoto, 860-0811, Japan

    Piyanard Boonnate, Kulthida Vaeteewoottacharn, Ryusho Kariya, Sawako Fujikawa & Seiji Okada

  2. Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Piyanard Boonnate & Kulthida Vaeteewoottacharn

  3. Department of Parasitology, Khon Kaen University, Khon Kaen, 40002, Thailand

    Thidarut Boonmars & Somchai Pinlaor

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

    Chawalit Pairojkul

  5. Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, 40002, Thailand

    Kulthida Vaeteewoottacharn, Thidarut Boonmars & Somchai Pinlaor

Authors
  1. Piyanard Boonnate
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  2. Kulthida Vaeteewoottacharn
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  3. Ryusho Kariya
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  4. Sawako Fujikawa
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  5. Thidarut Boonmars
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  6. Somchai Pinlaor
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  7. Chawalit Pairojkul
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  8. Seiji Okada
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Contributions

KV and SO conceived and designed the experiments. PB, KV, RK, and SF performed the experiments. PB. and RK. analyzed the data. PB, KV, and SO wrote the paper. All authors carefully read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Kulthida Vaeteewoottacharn or Seiji Okada.

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The authors declare no conflict of interest.

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11626_2021_608_MOESM1_ESM.tif

Figure S1 Cell cycle analysis of multinucleated Ham-1 and Ham-2 cells. Cells were stained with PI and were gated as shown from left to right. The multinucleated cells were the cells that have DNA content higher than G2 (> 4N) (tif 113 kb)

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Boonnate, P., Vaeteewoottacharn, K., Kariya, R. et al. Mucin-producing hamster cholangiocarcinoma cell line, Ham-2, possesses the aggressive cancer phenotypes with liver and lung metastases. In Vitro Cell.Dev.Biol.-Animal 57, 825–834 (2021). https://doi.org/10.1007/s11626-021-00608-z

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