This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Malignant transformation of the ovarian surface epithelium (OSE) accounts for most ovarian carcinomas. Detection of pre-neoplastic changes in the OSE leading to overt malignancy is important in prevention and management of ovarian cancer, especially in women with family histories of gynecological malignancies. The hypothesis of this study is that a spectrum of changes exists in the expression of individual OSE proteins that can eventually lead to overt carcinoma. OSE cell lines were obtained from women with and without familial ovarian cancer syndrome (FOCS) and known mutations in the tumor suppressor BRCA-1 gene. Lysates of OSE cell cultures were prepared and proteins were separated by isoelectric focusing, followed by SDS-PAGE. Gels were silver-stained, scanned and computer analyzed using 2D Analyzer Software. After matching gels, a protein database was created and proteins were characterized according to their molecular weight (MS), isoelectric point (pI) and relative abundance. Several different proteins were over-expressed, while others were under-expressed in OSE cells derived from women with FOCS. To identify these proteins, several unregulated proteins used as standards and the OSE proteins with altered expression in FOCS were excised and digested with trypsin. MALDI mass spectrometry was then performed and computerized database were identified by MALDI mass spectrometry include: 78 kDa glucose regulating protein, p22 calcium binding protein, ubiquitin carboxyl-terminal hydrolase, gamma-aminobutyric acid transaminase. MALDI masses of peptides drived from several altered OSE proteins have been obtained and identification based on computer database tryptic mass matching is in progress. The identification of these and other altered OSE proteins using mass spectrometry may be useful in detecting protein changes suggestive of FOCS, which may have potential as diagnostic cellular markers for pre-neoplastic ovarian disease.
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