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H-RYK, an Unusual Receptor Kinase: Isolation and Analysis of Expression in Ovarian Cancer

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Abstract

Background

Protein tyrosine kinases play an imporH-RYK, an Unusual Receptor Kinase: Isolation and Analysis of Expression in Ovarian Cancertant role in cellular metabolism as key components of signal transduction pathways. They are involved in cellular growth, differentiation, and development. Receptor tyrosine kinases (EGF receptor and c-erbB2) have been shown to be important in the pathogenesis of cancer. In ovarian cancer, overexpression of c-erbB2, a type I receptor, has been correlated with an adverse effect on survival of patients.

Material and Methods

An unusual receptor tyrosine kinase, H-RYK, has been isolated from a complimentary DNA library of SKOV-3, an epithelial ovarian cancer cell line, using a polymerase chain reaction-mediated approach.

Results

The primary structure of the predicted amino acid sequence of the protein shows a novel NH2-terminal region. The catalytic region shows homology to other tyrosine kinases, the closest homology being with v-sea(39%). A significant alteration in the catalytic domain is that the highly conserved “DFG” triplet in subdomain VII is altered to “DNA.” The gene was mapped to chromosome 3q22. A single transcript of 3.0 kb is expressed in heart, brain, lung, placenta, liver, muscle, kidney, and pancreas by Northern analysis with maximal expression in skeletal muscle. In situ hybridization analysis on human tissues demonstrated localization of message in the epithelial and stromal compartment of tissues such as brain, lung, colon, kidney, and breast. There was minimal to absent expression of H-RYK on surface epithelium of ovaries. In benign (3) and borderline tumors of the ovary (5), there was expression in the stromal compartment. However, in malignant tumors (24) there was increased expression predominantly confined to the epithelium. Polyclonal antisera raised against synthetic peptides recognize a 100-kD protein in ovarian cancer cells and other cell lines. In contrast to other receptor tyrosine kinases, the receptor did not phosphorylate in an in vitro kinase assay.

Conclusions

The expression of this unusual receptor tyrosine kinase in epithelial ovarian cancer suggests that it may be involved in tumor progression, which needs further investigation.

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Acknowledgments

We are grateful to Dr. J. Trowsdale and Dr. S. Chatterjee for the SKOV-3 and normal ovary cDNA libraries. Drs. A. P. Wilson and S. Langdon are gratefully acknowledged for the cell lines. ICRF Cell Production Services, In Situ Hybridization Service and Peptides Synthesis Laboratories are acknowledged. This work was supported by the Imperial Cancer Research Fund. Mr. R. Katso is a Rosina Valerie Howell Fellow.

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Authors and Affiliations

  1. ICRF Molecular Oncology Laboratories, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, OX3 9DU, UK

    Xiao C. Wang, Roy Katso, Robin Butler & Trivadi S. Ganesan

  2. RCS/ICRF Histopathology Unit, ICRF In Situ Hybridization Service, London, UK

    Andrew M. Hanby & Richard Poulsom

  3. Laboratory of Human Cytogenetics, London, UK

    Tania Jones & Denise Sheer

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  1. Xiao C. Wang
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  2. Roy Katso
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Wang, X.C., Katso, R., Butler, R. et al. H-RYK, an Unusual Receptor Kinase: Isolation and Analysis of Expression in Ovarian Cancer. Mol Med 2, 189–203 (1996). https://doi.org/10.1007/BF03401616

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