DNA damage repair is unaffected by mimicked heterozygous levels of BRCA2 in HT-29 cells.
Functional loss of both alleles of the breast cancer susceptibility gene, BRCA2, facilitates tumorigenesis. However, the direct effects of BRCA2 heterozygosity remain unclear. Here, BRCA2 heterozygosity was mimicked in HT-29 colon cells by reducing levels of BRCA2 through stable RNA interference. No difference in RAD51 subcellular localization and focus formation was observed between control and mimicked heterozygous cell lines. DNA repair ability, as measured by colony survival following mitomycin C treatment and ultraviolet radiation exposure, was also unaffected by reduced levels of BRCA2. Interestingly, the growth rate of the mimicked BRCA2 heterozygous cell line was significantly lower than that of control cells. Increased expression of p53 in the mimicked heterozygous cells was observed, perhaps in response to BRCA2 deficiency. Levels of p27 were also found to be slightly increased in cells with reduced BRCA2, perhaps contributing to the slower growth rate. Overall, these results suggest that tumors are unlikely to arise directly from BRCA2 heterozygous cells without other genetic events such as loss of the wild-type BRCA2 allele and/or loss of p53 function or other cell cycle inhibitors.
Tannenbaum
Brian
B
Department of Biology, Adelphi University, Garden City, NY 11530, USA.
Mofunanya
Tobechukwu
T
Schoenfeld
Alan R
AR
D024682:BRCA2 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D019073:HT29 Cells; D006579:Heterozygote; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D034622:RNA Interference; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D024682:BRCA2 Protein; D019384:Nucleic Acid Synthesis Inhibitors; D016159:Tumor Suppressor Protein p53; D050760:Cyclin-Dependent Kinase Inhibitor p27; D016685:Mitomycin