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Journal of Histochemistry and Cytochemistry, Vol. 49, 673-674, May 2001, Copyright © 2001, The Histochemical Society, Inc.

BRIEF REPORT

Monitoring Signal Transduction in Cancer: Tyrosine Kinase Gene Expression Profiling

Heinz-Ulrich G. Weiera, Horst F. Zitzelsbergerb, H.-Ben Hsieha, Melita V. Suna, Mariwil Wongc, Robert A. Lerscha, Paul Yaswena, Jan Smidab, Christine Kuschnickb, and Orlo H. Clarkc
a Life Sciences Division, University of California, E.O. Lawrence Berkeley National Laboratory, Berkeley, California
b Arbeitsgruppe Zytogenetik, gsf-Forschungszentrum fuer Umwelt und Gesundheit, Neuherberg, Germany
c Department of Surgery, University of California, San Francisco, California

Correspondence to: Heinz-Ulrich G. Weier, Life Sciences Division, Lawrence Berkeley National Laboratory, U. of California, 1 Cyclotron Road, MS 74–157, Berkeley, CA 94720. E-mail: ugweier@lbl.gov

Abnormal expression of tyrosine kinase (TK) genes is common in tumors, in which it is believed to alter cell growth and response to external stimuli such as growth factors and hormones. Although the etiology and pathogenesis of carcinomas of the thyroid or breast remain unclear, there is evidence that the expression of TK genes, such as receptor tyrosine kinases, or mitogen-activated protein kinases, is dysregulated in these tumors, and that overexpression of particular TK genes due to gene amplification, changes in gene regulation, or structural alterations leads to oncogenic transformation of epithelial cells. We developed a rapid scheme to measure semiquantitatively the expression levels of 50–100 TK genes. Our assay is based on RT-PCR with mixed based primers that anneal to conserved regions in the catalytic domain of TK genes to generate gene-specific fragments. PCR products are then labeled by random priming and hybridized to DNA microarrays carrying known TK gene targets. Inclusion of differently labeled fragments from reference or normal cells allows identification of TK genes that show altered expression levels during malignant transformation or tumor progression. Examples demonstrate how this innovative assay might help to define new markers for tumor progression and potential targets for disease intervention. (J Histochem Cytochem 49:673-674, 2001)

Key Words: tyrosine kinase, tumors, RT-PCR, genes


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