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[Cancer Research 59, 4142-4147, August 1, 1999]
© 1999 American Association for Cancer Research
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[Cancer Research 59, 4142-4147, August 15, 1999]
© 1999 American Association for Cancer Research

Tumor Biology

Intragenic Mutation Analysis of the Human Epidermal Growth Factor Receptor (EGFR) Gene in Malignant Human Oral Keratinocytes1

Satoru Shintani2, Kou Matsuo2, Constant C. Crohin, Jim McBride, Takanori Tsuji, R. Bruce Donoff, Marshall Posner, Randy Todd and David T.W. Wong3

Laboratory of Molecular Pathology, Division of Oral Pathology, Department of Oral Medicine and Diagnostic Sciences, Harvard University School of Dental Medicine [S. S., K. M., C. C. C., J. M., T. T., R. B. D., R. T., D. T. W. W.], and Head and Neck Oncology Group, Dana Farber Cancer Institute [M. P.], Boston, Massachusetts 02115

Alteration in epidermal growth factor receptor (EGFR) expression is frequently associated with malignant transformation of epithelial tissues, including oral mucosa. This study examines the mutations in the coding region of the human EGFR gene in normal and malignant human oral keratinocytes. To examine the intragenic mutations in the human EGFR gene, a panel of normal and malignant human oral keratinocytes were examined by a nonisotopic RNase cleavage assay. Two consistent alterations were detected. First, a polymorphism, which generates a unique BsrI restriction site, was detected at position 2073. This BsrI polymorphism was present only in malignant keratinocytes. Second, Southern blot hybridization of PCR products revealed that there is a truncated EGFR mRNA (~1.5-kb) in oral squamous cell carcinoma cell lines. Similar analysis in normal cell lines revealed that this truncated EGFR transcript is also present. Immunoblotting revealed the presence of this truncated form of EGFR in all keratinocyte cell lines. These data permit us to conclude that there exists a novel truncated form of EGFR in human oral keratinocytes. Furthermore, there exists a tumor-associated BsrI polymorphic site at position 2073. The potential biological relevance of the truncated receptor and the utility of the BsrI polymorphic site for diagnostic applications are currently being explored.




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Copyright © 1999 by the American Association for Cancer Research.