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

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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References

  1. Banales JM, Cardinale V, Carpino G, et al. Expert consensus document: cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA). Nat Rev Gastroenterol Hepatol. 2016;13:261–80.
    PubMed Google Scholar
  2. Thamavit W, Pairojkul C, Tiwawech D, Shirai T, Ito N. Strong promoting effect of Opisthorchis viverrini infection on dimethylnitrosamine-initiated hamster liver. Cancer Lett. 1994;78:121–5.
    CAS PubMed Google Scholar
  3. Parkin DM, Ohshima H, Srivatanakul P, Vatanasapt V. Cholangiocarcinoma: epidemiology, mechanisms of carcinogenesis and prevention. Cancer Epidemiol Biomark Prev. 1993;2:537–44.
    CAS Google Scholar
  4. Ohshima H, Bandaletova TY, Brouet I, et al. Increased nitrosamine and nitrate biosynthesis mediated by nitric oxide synthase induced in hamsters infected with liver fluke (Opisthorchis viverrini). Carcinogenesis. 1994;15:271–5.
    CAS PubMed Google Scholar
  5. Pinlaor S, Hiraku Y, Yongvanit P, et al. iNOS-dependent DNA damage via NF-kappaB expression in hamsters infected with Opisthorchis viverrini and its suppression by the antihelminthic drug praziquantel. Int J Cancer. 2006;119:1067–72.
    CAS PubMed Google Scholar
  6. Ben-Menachem T. Risk factors for cholangiocarcinoma. Eur J Gastroenterol Hepatol. 2007;19:615–7.
    PubMed Google Scholar
  7. Jinawath N, Chamgramol Y, Furukawa Y, et al. Comparison of gene expression profiles between Opisthorchis viverrini and non-Opisthorchis viverrini associated human intrahepatic cholangiocarcinoma. Hepatology. 2006;44:1025–38.
    CAS PubMed Google Scholar
  8. Jusakul A, Cutcutache I, Yong CH, et al. Whole-genome and epigenomic landscapes of etiologically distinct subtypes of cholangiocarcinoma. Cancer Discov. 2017;7:1116–35.
    CAS PubMed PubMed Central Google Scholar
  9. Dagogo-Jack I, Shaw AT. Tumour heterogeneity and resistance to cancer therapies. Nat Rev Clin Oncol. 2018;15:81–94.
    CAS PubMed Google Scholar
  10. DeOliveira ML, Cunningham SC, Cameron JL, et al. Cholangiocarcinoma: 31-year experience with 564 patients at a single institution. Ann Surg. 2007;245:755–62.
    PubMed PubMed Central Google Scholar
  11. Yusoff AR, Razak MM, Yoong BK, Vijeyasingam R, Siti ZM. Survival analysis of cholangiocarcinoma: a 10-year experience in Malaysia. World J Gastroenterol. 2012;18:458–65.
    PubMed PubMed Central Google Scholar
  12. Luvira V, Nilprapha K, Bhudhisawasdi V, Pugkhem A, Chamadol N, Kamsa-ard S. Cholangiocarcinoma patient outcome in northeastern thailand: single-center prospective study. Asian Pacif J Cancer Prev APJCP. 2016;17:401–6.
    Google Scholar
  13. Yonglitthipagon P, Pairojkul C, Chamgramol Y, Mulvenna J, Sripa B. Up-regulation of annexin A2 in cholangiocarcinoma caused by Opisthorchis viverrini and its implication as a prognostic marker. Int J Parasitol. 2010;40:1203–12.
    CAS PubMed PubMed Central Google Scholar
  14. Hahnvajanawong C, Ketnimit S, Pattanapanyasat K, et al. Involvement of p53 and nuclear factor-kappaB signaling pathway for the induction of G1-phase cell cycle arrest of cholangiocarcinoma cell lines by isomorellin. Biol Pharm Bull. 2012;35:1914–25.
    CAS PubMed Google Scholar
  15. Hunsawong T, Singsuksawat E, In-chon N, et al. Estrogen is increased in male cholangiocarcinoma patients' serum and stimulates invasion in cholangiocarcinoma cell lines in vitro. J Cancer Res Clin Oncol. 2012;138:1311–20.
    CAS PubMed Google Scholar
  16. Tepsiri N, Chaturat L, Sripa B, et al. Drug sensitivity and drug resistance profiles of human intrahepatic cholangiocarcinoma cell lines. World J Gastroenterol. 2005;11:2748–53.
    CAS PubMed PubMed Central Google Scholar
  17. Namwat N, Amimanan P, Loilome W, et al. Characterization of 5-fluorouracil-resistant cholangiocarcinoma cell lines. Chemotherapy. 2008;54:343–51.
    CAS PubMed Google Scholar
  18. Barr RJ, Hancock DE. Adenosquamous carcinoma of the liver. Gastroenterology. 1975;69:1326–30.
    CAS PubMed Google Scholar
  19. Nakanuma Y, Curado M, Franceschi S, Gore G, Paradis V, Sripa B. The International Agency for Research on Cancer, editors. Intrahepatic cholangiocarcioma. In: Bosman F, Carneiro F, Hruban R, Theise N, eds. WHO classification of tumours of the digestive system. Lyon IARC Press, 2010; 217–24.
  20. Sripa B, Leungwattanawanit S, Nitta T, et al. Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100). World J Gastroenterol. 2005;11:3392–7.
    PubMed PubMed Central Google Scholar
  21. Silsirivanit A, Araki N, Wongkham C, et al. A novel serum carbohydrate marker on mucin 5AC: values for diagnostic and prognostic indicators for cholangiocarcinoma. Cancer. 2011;117:3393–403.
    CAS PubMed Google Scholar
  22. An International System for Human Cytogenetic Nomenclature. Basel, Swizterland: Karger Medical and Scientific Publishers, 2013
  23. Phoomak C, Vaeteewoottacharn K, Sawanyawisuth K, et al. Mechanistic insights of O-GlcNAcylation that promote progression of cholangiocarcinoma cells via nuclear translocation of NF-kappaB. Scientific reports. 2016;6:27853.
    CAS PubMed PubMed Central Google Scholar
  24. Frankowski H, Gu YH, Heo JH, Milner R, Del Zoppo GJ. Use of gel zymography to examine matrix metalloproteinase (gelatinase) expression in brain tissue or in primary glial cultures. Methods Mol Biol. 2012;814:221–33.
    CAS PubMed PubMed Central Google Scholar
  25. Vaeteewoottacharn K, Kariya R, Dana P, et al. Inhibition of carbonic anhydrase potentiates bevacizumab treatment in cholangiocarcinoma. Tumour Biol. 2016;37:9023–35.
    CAS PubMed Google Scholar
  26. Eijssen LM, Jaillard M, Adriaens ME, et al. User-friendly solutions for microarray quality control and pre-processing on ArrayAnalysis.org. Nucleic acids Res. 2013; 41:W71–6.
  27. Eijssen LM, Goelela VS, Kelder T, Adriaens ME, Evelo CT, Radonjic M. A user-friendly workflow for analysis of Illumina gene expression bead array data available at the arrayanalysis.org portal. BMC Genomics. 2015;16:482.
  28. Saldanha AJ. Java Treeview–extensible visualization of microarray data. Bioinformatics. 2004;20:3246–8.
    CAS PubMed Google Scholar
  29. Suo Z, Holm R, Nesland JM. Squamous cell carcinomas. An immunohistochemical study of cytokeratins and involucrin in primary and metastatic tumours. Histopathology. 1993;23:45–54.
  30. Masters JR. False cell lines: the problem and a solution. Cytotechnology. 2002;39:69–74.
    CAS PubMed PubMed Central Google Scholar
  31. Markovic O, Markovic N. Cell cross-contamination in cell cultures: the silent and neglected danger vitro cellular and developmental biology. Animal. 1998;34:1–8.
    CAS Google Scholar
  32. MacLeod RA, Dirks WG, Matsuo Y, Kaufmann M, Milch H, Drexler HG. Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int J Cancer. 1999;83:555–63.
    CAS PubMed Google Scholar
  33. Azari S, Ahmadi N, Tehrani MJ, Shokri F. Profiling and authentication of human cell lines using short tandem repeat (STR) loci: report from the National Cell Bank of Iran. Biologicals. 2007;35:195–202.
    CAS PubMed Google Scholar
  34. Lacroix M. Persistent use of "false" cell lines. Int J Cancer. 2008;122:1–4.
    CAS PubMed Google Scholar
  35. Horbach S, Halffman W. The ghosts of HeLa: How cell line misidentification contaminates the scientific literature. PLoS ONE. 2017;12:e0186281.
    PubMed PubMed Central Google Scholar
  36. Masters JR. HeLa cells 50 years on: the good, the bad and the ugly. Nat Rev Cancer. 2002;2:315–9.
    CAS PubMed Google Scholar
  37. Petljak M, Alexandrov LB, Brammeld JS, et al. Characterizing mutational signatures in human cancer cell lines reveals episodic APOBEC mutagenesis. Cell. 2019;176(1282–94):e20.
    Google Scholar
  38. Ono I, Ishiwata I, Tashiro A, et al. Establishment and characterization of two human mixed mesodermal tumor cell lines from the same patient. J Natl Cancer Inst. 1984;72:1241–52.
    CAS PubMed Google Scholar
  39. Navone NM, Olive M, Ozen M, et al. Establishment of two human prostate cancer cell lines derived from a single bone metastasis. Clin Cancer Res. 1997;3:2493–500.
    CAS PubMed Google Scholar
  40. Enjoji M, Sakai H, Nawata H, Kajiyama K, Tsuneyoshi M. Sarcomatous and adenocarcinoma cell lines from the same nodule of cholangiocarcinoma vitro cellular and developmental biology. Animal. 1997;33:681–3.
    CAS Google Scholar
  41. Nicholson JM, Cimini D. Cancer karyotypes: survival of the fittest. Front Oncol. 2013;3:148.
    PubMed PubMed Central Google Scholar
  42. Ben-David U, Siranosian B, Ha G, et al. Genetic and transcriptional evolution alters cancer cell line drug response. Nature. 2018;560:325–30.
    CAS PubMed PubMed Central Google Scholar
  43. Rattanasinganchan P, Leelawat K, Treepongkaruna SA, et al. Establishment and characterization of a cholangiocarcinoma cell line (RMCCA-1) from a Thai patient. World J Gastroenterol. 2006;12:6500–6.
    CAS PubMed PubMed Central Google Scholar
  44. Ma S, Hu L, Huang XH, et al. Establishment and characterization of a human cholangiocarcinoma cell line. Oncol Rep. 2007;18:1195–200.
    PubMed Google Scholar
  45. Cavalloni G, Peraldo-Neia C, Varamo C, et al. Establishment and characterization of a human intrahepatic cholangiocarcinoma cell line derived from an Italian patient. Tumour Biol. 2016;37:4041–52.
    CAS PubMed Google Scholar
  46. Saensa-Ard S, Leuangwattanawanit S, Senggunprai L, et al. Establishment of cholangiocarcinoma cell lines from patients in the endemic area of liver fluke infection in Thailand. Tumour Biol. 2017;39:1010428317725925.
    PubMed Google Scholar
  47. Dirks WG, Faehnrich S, Estella IA, Drexler HG. Short tandem repeat DNA typing provides an international reference standard for authentication of human cell lines. Altex. 2005;22:103–9.
    PubMed Google Scholar
  48. Yoshino K, Iimura E, Saijo K, et al. Essential role for gene profiling analysis in the authentication of human cell lines. Hum Cell. 2006;19:43–8.
    PubMed Google Scholar
  49. Almeida JL, Cole KD, Plant AL. Standards for cell line authentication and beyond. PLoS Biol. 2016;14:e1002476.
    PubMed PubMed Central Google Scholar
  50. Sirisinha S, Tengchaisri T, Boonpucknavig S, Prempracha N, Ratanarapee S, Pausawasdi A. Establishment and characterization of a cholangiocarcinoma cell line from a Thai patient with intrahepatic bile duct cancer. Asian Pac J Allergy Immunol. 1991;9:153–7.
    CAS PubMed Google Scholar
  51. Vaeteewoottacharn K, Pairojkul C, Kariya R, et al. Establishment of highly transplantable cholangiocarcinoma cell lines from a patient-derived xenograft mouse model. Cells. 2019;8:496–510.
    CAS PubMed Central Google Scholar

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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

Authors

  1. Banchob Sripa
  2. Wunchana Seubwai
  3. Kulthida Vaeteewoottacharn
  4. Kanlayanee Sawanyawisuth
  5. Atit Silsirivanit
  6. Worasak Kaewkong
  7. Kanha Muisuk
  8. Paweena Dana
  9. Chatchai Phoomak
  10. Worachart Lert-itthiporn
  11. Vor Luvira
  12. Chawalit Pairojkul
  13. Bin T. Teh
  14. Sopit Wongkham
  15. Seiji Okada
  16. Yaovalux Chamgramol

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Correspondence toSeiji 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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