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Functional and genetic characterization of three cell lines derived from a single tumor of an Opisthorchis viverrini-associated cholangiocarcinoma patient

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Abstract

Three cholangiocarcinoma (CCA) cell line—formerly named, M156, M213 and M214 have been intensively used with discrepancy of their tumor origins. They were assumed to be originated from three different donors without authentication. To verify the origins of these cell lines, the short tandem repeat (STR) analysis of the currently used cell lines, the cell stocks from the establisher and the primary tumor of a CCA patient were performed. Their phenotypic and genotypic originality were compared. The currently used 3 CCA cell lines exhibited similar STR as CCA patient ID-M213 indicating the same origin of these cells. The cell stocks from the establisher, however, revealed the same STR of M213 and M214 cells, but not M156. The misidentification of M214 and M156 is probably due to the mislabeling and cross-contamination of M213 cells during culture. These currently used cell lines were renamed as KKU-213A, -213B and -213C, for the formerly M213, M214 and M156 cells, respectively. These cell lines were established from a male with an intrahepatic mass-forming CCA stage-4B. The tumor was an adenosquamous carcinoma with the liver fluke ova granuloma in evidence. All cell lines had positive CK19 with differential CA19-9 expression. They exhibited aneuploidy karyotypes, distinct cell morphology, cell growth, cytogenetic characteristic and progressive phenotypes. KKU-213C formed a adenosquamous carcinoma, whereas KKU-213A and KKU-213B formed poorly- and well-differentiated squamous cell carcinomas in xenografted mice. mRNA microarray revealed different expression profiles among these three cell lines. The three cell lines have unique characteristics and may resemble the heterogeneity of tumor origin.

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Acknowledgements

This work was co-supported by research grants from the Khon Kaen University (I6201-02) and the Thailand Research Fund (DBG5980004). The authors would like to thank D. Jefferson (New England Medical Center, Tufts University) for the H69 cell line and Prof. James A. Will for editing this manuscript via the Faculty of Medicine Publication Clinic, Khon Kaen University. We would also express our sincere thanks to the reviewers and editor of Human Cell for their critical advices through the preparation of this manuscript.

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Authors and Affiliations

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

    Banchob Sripa, Chawalit Pairojkul & Yaovalux Chamgramol

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

    Wunchana Seubwai & Kanha Muisuk

  3. Department of Biochemistry, and Center for Translational Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Kulthida Vaeteewoottacharn, Kanlayanee Sawanyawisuth, Atit Silsirivanit, Paweena Dana, Chatchai Phoomak, Worachart Lert-itthiporn & Sopit Wongkham

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

    Vor Luvira

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

    Banchob Sripa, Wunchana Seubwai, Kulthida Vaeteewoottacharn, Kanlayanee Sawanyawisuth, Atit Silsirivanit, Worasak Kaewkong, Kanha Muisuk, Paweena Dana, Chatchai Phoomak, Worachart Lert-itthiporn, Chawalit Pairojkul, Sopit Wongkham & Yaovalux Chamgramol

  6. Department of Biochemistry, Faculty of Medical Sciences, Naresuan University, Phitsanulok, 65000, Thailand

    Worasak Kaewkong

  7. Duke-NUS Medical School, Singapore, 169857, Singapore

    Bin T. Teh

  8. Division of Hematopoeisis, Joint Research Center for Human Retrovirus Infection, Kumamoto University, Kumamoto, 860-0811, Japan

    Seiji Okada

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  1. Banchob Sripa
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Correspondence to Seiji Okada or Yaovalux Chamgramol.

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Sripa, B., Seubwai, W., Vaeteewoottacharn, K. et al. Functional and genetic characterization of three cell lines derived from a single tumor of an Opisthorchis viverrini-associated cholangiocarcinoma patient. Human Cell 33, 695–708 (2020). https://doi.org/10.1007/s13577-020-00334-w

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