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Establishment and characterization of a platinum- and paclitaxel-resistant high grade serous ovarian carcinoma cell line

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Abstract

High grade serous ovarian cancer (HGSOC) patients have a high recurrence rate after surgery and adjuvant chemotherapy due to inherent or acquired drug resistance. Cell lines derived from HGSOC tumors that are resistant to chemotherapeutic agents represent useful pre-clinical models for drug discovery. Here, we describe establishment of a human ovarian carcinoma cell line, which we term WHIRC01, from a patient-derived mouse xenograft established from a chemorefractory HGSOC patient who did not respond to carboplatin and paclitaxel therapy. This newly derived cell line is platinum- and paclitaxel-resistant with cisplatin, carboplatin, and paclitaxel half-maximal lethal doses of 15, 130, and 20 µM, respectively. Molecular characterization of this cell line was performed using targeted DNA exome sequencing, transcriptomics (RNA-seq), and mass spectrometry-based proteomic analyses. Results from exomic sequencing revealed mutations in TP53 consistent with HGSOC. Transcriptomic and proteomic analyses of WHIRC01 showed high level of alpha-enolase and vimentin, which are associated with cell migration and epithelial–mesenchymal transition. WHIRC01 represents a chemorefractory human HGSOC cell line model with a comprehensive molecular profile to aid future investigations of drug resistance mechanisms and screening of chemotherapeutic agents.

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Acknowledgements

This research was supported by the Uniformed Services University of the Health Sciences under Award Number HU0001-16-2-0006. We would like to acknowledge Kevin D. Stroop for his assistance in sample preparation.

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Authors and Affiliations

  1. Women’s Health Integrated Research Center at Inova Health System, Gynecologic Cancer Center of Excellence, Annandale, VA, USA

    Pang-Ning Teng, Nicholas W. Bateman, Guisong Wang, Tracy Litzi, Brian E. Blanton, Brian L. Hood, Kelly A. Conrads, Wei Ao, Kathleen M. Darcy, Chad A. Hamilton, G. Larry Maxwell & Thomas P. Conrads

  2. The John P. Murtha Cancer Center, Walter Reed National Military Medical Center, Bethesda, MD, USA

    Nicholas W. Bateman, Kathleen M. Darcy, Chad A. Hamilton, G. Larry Maxwell & Thomas P. Conrads

  3. Gynecologic Oncology Service, Department of Obstetrics and Gynecology, Walter Reed National Military Medical Center, Bethesda, MD, USA

    Kate E. Oliver & Chad A. Hamilton

  4. Department of Obstetrics and Gynecology, Inova Fairfax Hospital, Falls Church, VA, USA

    G. Larry Maxwell & Thomas P. Conrads

  5. Department of Internal Medicine, Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA

    William P. McGuire

  6. Champions Oncology, Inc., Baltimore, MD, USA

    Keren Paz

  7. Otolaryngology-Head and Neck Surgery and Oncology, Johns Hopkins University, Baltimore, MD, USA

    David Sidransky

  8. Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, MD, USA

    Nicholas W. Bateman & Thomas P. Conrads

  9. Inova Schar Cancer Institute, Inova Center for Personalized Health, Falls Church, VA, USA

    G. Larry Maxwell & Thomas P. Conrads

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  1. Pang-Ning Teng
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Supplementary Table 1. Mutation profile of WHIRC01

Supplementary Table 2. WHIRC01 transcriptome

Supplementary Table 3. Morphological subtype gene list

Supplementary Table 4. TCGA ovarian cancer tumor cohort

Supplementary Table 5. Molecular subtype gene list

Supplementary Table 6. WHIRC01 and OV90 proteome

Supplementary Fig. 1 Confirmation of human origin of WHIRC01 cell line by CO1 PCR Assay. WHIRC01 was verified to be human by the presence of the human CO1 (391 bp) band and the absence of mouse CO1 (150 bp) band in the CO1 PCR assay.

Supplementary Fig. 2 WHIRC01 growth curve and cell doubling time. Average live cells counts were used to generate the growth curve of WHIRC01. The slope of the exponential portion (day 3 – 7) of the growth curve was used to calculate the cell doubling time of WHIRC01 to be approximately 30 h.

Supplementary Fig. 3 Inhibition of ATR sensitized WHIRC01 to Carboplatin. Dose response of WHIRC01 to carboplatin (72 h treatment) in the presence or absence of ATR inhibitor (AZD6738, 1 µM) was measured by the MTS assay. AZD6738 sensitized WHIRC01 to carboplatin.

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Teng, PN., Bateman, N.W., Wang, G. et al. Establishment and characterization of a platinum- and paclitaxel-resistant high grade serous ovarian carcinoma cell line. Human Cell 30, 226–236 (2017). https://doi.org/10.1007/s13577-017-0162-1

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