Considerable circumstantial evidence suggests that oncogenes play a role in the development and/or progression of human malignancies. Very little work has been done with human gynecologic neoplasms to test this concept. Our preliminary results show that the HER-2/neu proto-oncogene is amplified and overexpressed in human ovarian cancer and in endometrial cancer. Amplification and expression of the HER-2/neu oncogene will be characterized with Southern hybridization, Northern hybridization, Western immunoblot analysis and immunohistochemistry to determine if amplification is associated with overexpression. Gene amplification will be compared with messenger RNA levels, oncoprotein levels and tissue/cellular distribution of oncoprotein. A computerized image analysis method for quantification of HER-2/neu oncoprotein in individual cells will be developed and used in this investigation. The DNA content of the cancers, considered to be a prognosidc factor in ovarian cancer, will also be determined. Clinical parameters, especially overall patient survival, will be obtained and compared with oncogene amplification and expression as well as DNA ploidy content to determine if there is any correlation between oncoprotein expression, DNA ploidy and unfavorable clinical outcome in women with ovarian or endometrial cancer. In our preliminary data 26% of ovarian cancers had amplification of this oncogene and amplification correlated with a shortened overall patient survival. Since HER-2/neu may be of diagnostic and potentially of therapeutic utility its distribution and expression in normal tissues is critically important. This information is currently unavailable and will be determined as part of the investigation. What role, if any, that wild-type (unmutated) HER-2/neu may play in the pathogenesis of ovarian and/or endometrial cancers is unknown. The transmembrane domain of HER-2/neu gene will be isolated with the polymerase chain reaction and sequenced to determine if the activating point mutation present in mouse tumors is present in human human overian and endometrial cancers. Wild-type HER-2/neu gene will be injected into mousc embryos to produce transgenic mice to determine if this amplified gene is sufficient for the production of malignant tumors. The long-term objective of the current proposal is to determine if the HER-2/neu proto-oncogene is clinically important in human gynecologic neoplasms and to understand the mechanisms by which this cellular oncogene regulates the metabolism of normal and malignant cells of the female reproductive tract
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