Mutations in tumor-suppressor gene BARD1 have been found in inherited and spontaneous breast, ovarian and uterine cancers. BARD1 plays a critical role in DNA repair and ubiquitination as binding partner of BRCA1, with which it colocalizes to nuclear dots. Independently of BRCA1, BARD1 can induce p53-dependent apoptosis in response to genotoxic stress. Therefore, BARD1 or p53 might be defective in cancer cells spared from apoptosis. We investigated BARD1 and p53 expression in ovarian, breast and non-small-cell lung cancers. BARD1 expression was highly upregulated and cytoplasmic in most cancer cells, while weak nuclear staining was observed in the surrounding normal tissue. Maximal BARD1 expression was associated with the most malignant ovarian cancer, clear cell carcinoma. In breast cancer, BARD1 expression was correlated with poor differentiation and large tumor size, established factors of poor prognosis, as well as short disease-free survival. In contrast to breast and ovarian cancers, no correlation of BARD1 expression with either grade or stage could be determined for lung cancer. RT-PCR, performed on 10 ovarian cancers, revealed absence of the 5 0 portion of the BARD1 transcript in 7 tumors, and sequencing of the remaining 3 identified a missense mutation (A1291G) resulting in an amino acid change of glutamine 406 to arginine. These data suggest that genetic and epigenetic changes might lead to elevated cytoplasmic expression of BARD1 and that cytoplasmic BARD1 might be a poor prognostic factor for breast and ovarian cancers. ' 2005 Wiley-Liss, Inc.Key words: BARD1; BRCA1; breast cancer; lung cancer; ovarian cancer; p53Cancer has become the second leading cause of death in Western countries. A total of 1,368,030 new cancer cases and 563,700 deaths are expected in the United States in 2004. Although lung cancer is the leading cause of cancer death for men, breast and ovarian cancers account for approximately 32% and 4% of cases in women, respectively. 1 Ninety percent of malignant primary ovarian neoplasms are surface epithelial-stromal tumors, and malignant germ cell or sexcord stroma tumors are less common. Serous, mucinous, endometrioid and clear cell adenocarcinomas are more frequent than transitional cell tumors, mixed epithelial and undifferentiated carcinomas. 1a Of these subtypes, clear cell carcinoma is reported to have the worst prognosis. 2 Only a few prognostic factors, although controversial, have been identified in ovarian cancer so far, including cyclin D, p53, p21, and CA125. 3,4 The frequency of overexpression of mutant p53 is significantly higher in advanced-stage (III/IV) ovarian cancer compared to stage I cases (10-20%). This may indicate that p53 inactivation is a late event in ovarian carcinogenesis. [5][6][7] More factors have been reported to influence the prognosis for breast cancer, including size, histologic type and grade of the primary tumor, lymph node involvement, status of estrogen and progesterone receptors and other biomarkers such as HER-2, p53, bcl-2, Bfl-1, Ki-67 and VEG...