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In vitro detection of cholangiocarcinoma cells using a fluorescent protein-expressing oncolytic herpes virus

Cancer Gene Therapy volume 24, pages 227–232 (2017)Cite this article

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Abstract

Pathological confirmation is desired prior to high-risk surgery for suspected perihilar cholangiocarcinoma (PHC), but preoperative tissue diagnosis is limited by poor sensitivity of available techniques. This study aimed to validate whether a tumor-specific enhanced green fluorescent protein (eGFP)-expressing oncolytic virus could be used for cholangiocarcinoma (CC) cell detection. Extrahepatic CC cell lines SK-ChA-1, EGI-1, TFK-1 and control cells (primary human liver cells) were exposed to the oncolytic herpes simplex type 1 virus NV1066 for up to 24 h in adherent culture. The technique was validated for cells in suspension and cultured cells that had been exposed to crude patient bile. Optimal incubation time of the CC cells with NV1066 at a multiplicity of infection of 0.1 was determined at 6–8 h, yielding 15% eGFP-expressing cells, as measured by flow cytometry. Cells were able to survive 2-h crude bile exposure and remained capable of producing eGFP following NV1066 infection. Detection of malignant cells was possible at the highest dilution tested (10 CC cells among 2 × 105 control cells), though hampered by non-target cell autofluorescence. The technique was not applicable to cells in suspension due to insufficient eGFP production. Accordingly, as yet the technique is not suitable for standardized clinical diagnostics in PHC.

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Acknowledgements

We are grateful to Dr NA Kootstra and A van Nuenen for providing facility for viral culturing. We thank B Hooijbrink for assisting in the flow cytometry analysis and Prof. S Repping for providing the Vero cells. The SK-ChA-1 cells were a kind gift from A Knuth and C Matter from the University Hospital Zurich, Switzerland. The EGI-1 and TFK-1 cells were provided by Prof. RP Oude Elferink from the Tytgat Institute for Liver and Intestinal Research, Department of Gastroenterology and Hepatology, Academic Medical Center.

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Author notes

  1. R J S Coelen and M J de Keijzer: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Experimental Surgery, Academic Medical Center, Amsterdam, The Netherlands

    R J S Coelen, M J de Keijzer, R Weijer, V V Loukachov, J K Wiggers, A C W A van Wijk, M Heger & T M van Gulik

  2. Department of Research Facilities, Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    F P J Mul

  3. Department of Surgery, City of Hope, Duarte, CA, USA

    Y Fong

Authors
  1. R J S Coelen
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  2. M J de Keijzer
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  3. R Weijer
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  4. V V Loukachov
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  5. J K Wiggers
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  6. F P J Mul
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  7. A C W A van Wijk
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  8. Y Fong
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  9. M Heger
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  10. T M van Gulik
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Corresponding author

Correspondence to R J S Coelen.

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The authors declare no conflict of interest.

Additional information

This study was previously presented at the 51st Congress of the European Society for Surgical Research, 25–28 May 2016, Prague, Czech Republic.

Supplementary Information accompanies the paper on Cancer Gene Therapy website

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Coelen, R., de Keijzer, M., Weijer, R. et al. In vitro detection of cholangiocarcinoma cells using a fluorescent protein-expressing oncolytic herpes virus. Cancer Gene Ther 24, 227–232 (2017). https://doi.org/10.1038/cgt.2017.11

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