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Sequential genetic change at the TP53 and chemokine receptor CXCR4 locus during transformation of human ovarian surface epithelium

Oncogene volume 31, pages 4987–4995 (2012)Cite this article

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Abstract

Early genetic events in the development of high-grade serous ovarian cancer (HGSOC) may define the molecular basis of the profound structural and numerical instability of chromosomes in this disease. To discover candidate genetic changes we sequentially passaged cells from a karyotypically normal hTERT immortalised human ovarian surface epithelial line (IOSE25) resulting in the spontaneous formation of colonies in soft agar. Cell lines transformed ovarian surface epithelium 1 and 4 (TOSE 1 and 4) established from these colonies had an abnormal karyotype and altered morphology, but were not tumourigenic in immunodeficient mice. TOSE cells showed loss of heterozygosity (LOH) at TP53, increased nuclear p53 immunoreactivity and altered expression profile of p53 target genes. The parental IOSE25 cells contained a missense, heterozygous R175H mutation in TP53, whereas TOSE cells had LOH at the TP53 locus with a new R273H mutation at the previous wild-type TP53 allele. Cytogenetic and array CGH analysis of TOSE cells also revealed a focal genomic amplification of CXCR4, a chemokine receptor commonly expressed by HGSOC cells. TOSE cells had increased functional CXCR4 protein and its abrogation reduced epidermal growth factor receptor (EGFR) expression, as well as colony size and number. The CXCR4 ligand, CXCL12, was epigenetically silenced in TOSE cells and its forced expression increased TOSE colony size. TOSE cells had other cytogenetic changes typical of those seen in HGSOC ovarian cancer cell lines and biopsies. In addition, enrichment of CXCR4 pathway in expression profiles from HGSOC correlated with enrichment of a mutated TP53 gene expression signature and of EGFR pathway genes. Our data suggest that mutations in TP53 and amplification of the CXCR4 gene locus may be early events in the development of HGSOC, and associated with chromosomal instability.

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Acknowledgements

We thank Nitzan Rosenfeld and Davina Gale for their expert advice and analysis for the TP53 quantification. We also thank Debra Lillington for karyotyping TOSE cells, and the Genome Centre at Barts and the London School of Medicine and Dentistry for microsatellite analysis. This work was supported by Cancer Research UK, the Association for International Cancer Research and HEFCE.

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Authors and Affiliations

  1. Centre for Cancer and Inflammation, Barts Cancer Institute, Queen Mary University of London, London, UK

    K M Archibald, H Kulbe & F Balkwill

  2. Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Hong Kong, Republic of China

    J Kwong

  3. Bioinformatics and Biostatistics Service, London, UK

    P Chakravarty

  4. Functional Genomics of Ovarian Cancer Laboratory, Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Cambridge, UK

    J Temple & J D Brenton

  5. Centre for Haemato-Oncology, Barts Cancer Institute, Queen Mary University of London, London, UK

    T Chaplin & B D Young

  6. Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, London, UK

    M B Flak & I A McNeish

  7. Department of Histopathology, University Hospitals Nottingham, Nottingham, UK

    S Deen

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  1. K M Archibald
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Correspondence to F Balkwill.

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Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Microsatellite analysis of cell lines (PDF 27 kb)

List of differentially expressed p53 target genes between IOSE25 and TOSE1 (PDF 76 kb)

Sanger sequencing of exons 2-11 of TP53 in IOSE and TOSE cells (PDF 60 kb)

41388_2012_BFonc2011653_MOESM5_ESM.pdf

Cytogenetic gains in TOSE cells correlated with ovarian cancer cell lines and with published associations in human ovarian cancer samples (PDF 45 kb)

41388_2012_BFonc2011653_MOESM6_ESM.pdf

Cytogenetic gains in TOSE cells correlated with ovarian cancer cell lines and with published associations in human ovarian cancer samples (PDF 42 kb)

CXCL12/CXCR4 pathway genes (XLS 34 kb)

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Archibald, K., Kulbe, H., Kwong, J. et al. Sequential genetic change at the TP53 and chemokine receptor CXCR4 locus during transformation of human ovarian surface epithelium. Oncogene 31, 4987–4995 (2012). https://doi.org/10.1038/onc.2011.653

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