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GPI-specific phospholipase D mRNA expression in tumor cells of different malignancy

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Abstract

Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is a highly specific enzyme whose only known substrate is the GPI anchor of cell surface proteins. GPI-PLD measurements, however, are technically difficult since the enzyme is expressed at low levels in cells and tissues, and serum contains large amounts of inactive, latent GPI-PLD interfering with protein-based assays. We have therefore developed a semi-quantitative RT-PCR method to measure mRNA expression of all known GPI-PLD isoforms in cells and tissues. In human ovarian cancer cell lines, GPI-PLD mRNA expression correlated with GPI-PLD enzyme activity and with the shedding of the GPI-anchored tumor and prognostic markers, urokinase receptor and CA125, from the cell surface. This supports a potential role for this enzyme in the generation of circulating prognostic markers in malignant tumors. Similarly, in human epithelial cells of the skin, GPI-PLD mRNA expression increased with tumor progression. Whereas normal keratinocytes did not express significant amounts of GPI-PLD mRNA, expression was dramatically induced by serum in immortalized HaCaT keratinocytes and constitutively high and independent of serum in tumorigenic A431 epidermoid carcinoma cells. In addition, GPI-PLD expression was significantly increased in highly malignant, H-ras-transfected murine bladder carcinoma cells as compared to the low malignant, non-transfected parental cells. The competitive RT-PCR described here represents the first quantitative assay specific for cellular GPI-PLD isoforms, and our in vitro analyses suggest that GPI-PLD expression might be associated with tumor malignancy.

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Authors and Affiliations

  1. Department of Cancer Research, Fachklinik Hornheide, University of Münster, Germany

    Xiaotong He & Melanie-Jane Hannocks

  2. School of Biological Sciences, University of Manchester, UK

    Xiaotong He & Georg Brunner

  3. Department of Medicine, University of Manchester, UK

    Ian Hampson

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  1. Xiaotong He
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  2. Melanie-Jane Hannocks
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  3. Ian Hampson
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  4. Georg Brunner
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Correspondence to Georg Brunner.

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He, X., Hannocks, MJ., Hampson, I. et al. GPI-specific phospholipase D mRNA expression in tumor cells of different malignancy. Clin Exp Metastasis 19, 291–299 (2002). https://doi.org/10.1023/A:1015545407700

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