Proteome changes in ovarian epithelial cells derived from women with BRCA1 mutations and family histories of cancer.
Malignant transformation of the ovarian surface epithelium (OSE) accounts for most ovarian carcinoma. Detection of preneoplastic changes in the OSE leading to overt malignancy is important in prevention and management of ovarian cancer. We identified OSE proteins with altered expression derived from women with a family history (FH) of ovarian and/or breast cancer and mutations in the BRCA1 tumor suppressor gene. Proteins from SV-40-transformed FH-OSE cell lines and control OSE lines derived from women without such histories (non-family history) were separated by two-dimensional PAGE. Gels were analyzed, a protein data base was created, and proteins were characterized according to their molecular weight, isoelectric point, and relative abundance. Mass spectrometry was performed on tryptic protein digests, and data bases were searched for known proteins with the same theoretical tryptic peptide masses. Several proteins showed altered expression in the FH-OSE cells. Beta-tubulin and to a lesser extent ubiquitin carboxyl-terminal hydrolase and glyoxalase 1 appeared to be up-regulated. In contrast, proteins suppressed in FH lines include the 27-kDa heat shock protein, translationally controlled tumor protein, and several proteins associated with actin modification such as actin prepeptide, F-actin capping protein alpha subunit, and cofilin. Sequencing of several cofilin gel spots revealed phosphorylation of serine 3, a post-translational modification associated with decreased actin binding and cytoskeletal reorganization. Two-dimensional Western blots probed with cofilin antibody showed multiple protein spots with isoelectric points of 6-9 pH units. Blots of one-dimensional gels showed a significant reduction in cofilin expression in three FH lines when compared with three non-family history lines (p < or = 0.05). Identification of these and other OSE proteins may be useful in detecting changes suggestive of increased risk of developing preneoplastic disease and defining the possible role(s) of the BRCA1 gene in regulation of OSE cell function.
Smith-Beckerman
Diana M
DM
Center for Biomedical Laboratory Science, Biology Department, San Francisco State University, 1600 Holloway Avenue, San Francisco, CA 94132, USA. dmsmithb@sfsu.edu
Fung
Kit W
KW
Williams
Katherine E
KE
Auersperg
Nelly
N
Godwin
Andrew K
AK
Burlingame
Alma L
AL
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000737:chemistry; D000465:Algorithms; D000595:Amino Acid Sequence; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism*; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000378:metabolism*; D005192:Family Health; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D019398:Genes, BRCA1; D006801:Humans; D015151:Immunoblotting; D007525:Isoelectric Focusing; D007791:Lactoylglutathione Lyase / Q000096:biosynthesis; D013058:Mass Spectrometry; D008840:Microfilament Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D009369:Neoplasms / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology; D025801:Ubiquitin / Q000737:chemistry; D015854:Up-Regulation
D051339:Actin Depolymerizing Factors; D000199:Actins; D008840:Microfilament Proteins; D010455:Peptides; D025801:Ubiquitin; D014357:Trypsin; D007791:Lactoylglutathione Lyase