Antisense Inhibition of BRCA1 Expression and Molecular Analysis of Hereditary Tumors Indicate That Functional Inactivation of the p53 DNA Damage Response Pathway Is Required for BRCA-Associated Tumorigenesis

Abstract Objective. The purpose of this investigation was to test the hypothesis that mutation of TP53 is a requirement for BRCA-associated cancer development. Methods. A cell line experimentally deficient in BRCA1 protein was constructed using a regulatable antisense expression vector expressing 4000 bp from the BRCA1 cDNA. Changes in BRCA1, p53, and p21 protein levels were assayed by immunoblotting. Ovarian tumors with germline mutations in BRCA1 or BRCA2 were screened for mutations in TP53 by single-strand conformation polymorphism analysis. Results. Antisense inhibition of BRCA1 protein caused p53 and p21 protein levels to rise, indicating that partial loss of BRCA1 function activates the p53 DNA damage response pathway. Somatic mutation of TP53 was observed in 7 of 14 BRCA-associated ovarian tumors. Conclusions. Our findings provide novel evidence that loss of BRCA1 function in human cells activates the p53 DNA damage response pathway and that loss of this pathway, by somatic mutation of TP53, is a likely requirement for BRCA-associated tumor development.