Mutation Detection in EXON 11 of BRCAI in Filipino Breast Cancer Patients by PCR-SSCP

Abstract

The BRCA1 is a tumor suppressor gene which when mutated can predispose an individual to breast cancer. DNA was extracted from the blood of 17 subjects, 12 of whom comprised the patient population and the other 5 comprised the control population. The region in exon 11 of BRCA1, where a unique Filipino mutation was previously reported by Worsham and colleagues (1998), was amplified through polymerase chain reaction (PCR) with an annealing temperature of 68°C with 200 ng DNA template. Single stranded conformation polymorphism (SSCP) was carried out in 8% (w/v) polyacrylamide at 4°C for approximately 8 hours. For the first batch of SSCP, 81.57% of the patient population exhibited a different banding profile from that of the controls, thus suggesting the probable presence of mutations. For the second batch of SSCP, 100% of the patients exhibited a different banding profile from that of the controls, thus suggesting the possible presence of mutations. The presence and nature of these putative mutations can only be confirmed by gene sequencing, which is beyond the scope of this study.