| [Objective] The BRCA1 gene is a breast cancer Susceptibility gene and its germline mutation makes the risk of female breast cancer increased significantly. In order to more fully determine the BRCA1 gene in breast cancer related to the distribution of population, the study is mainly on detecting and analysis cancer BRCA1 gene mutations and rearrangements in population related breast cancer. Meanwhile, this study is to explore the relationship between the BRCA1 gene mutation and rearrangement mechanism, and between breast cancer and its impact on the significance of early diagnosis of breast cancer.[Methods] During the time from December, 2006 to January, 2008, Chooses breast cancer relative case sample altogether 99 examples from Shenzhen mother and child care courtyard mammary gland surgical department, including pathology diagnosis breast cancer sample 63 examples and breast cancer 1 level of relative sample 36 examples. Normal comparison group are BRCA1 gene wild-type samples. Screening 99 cancer relative case sampless BRCA1 gene . then have the unusual peak samples been carried on the direct sequence. Comparing the results with sequence from the Gene Bank, thus examines the sudden change of the BRCA1 gene; Using the technology of Multiplex ligation-dependent probe amplification to examine BRCA1 gene of 99 example breast cancer related crowd for big fragment delete and the redundant. [Result]1. Screening 99 peripheral blood genomic DNA BRCA1 gene for cancer-causing mutations with PCR-dHPLC technology, and found that six kinds of abnormal DHPLC peak, which were sequenced by DNA and conclude that four kinds are of non-sense mutation, which are Exon3 CD38AAG→AAA(Lys→Lys),Exon11 CD694 AGC→AGT(Ser→Ser),Exon11 CD771 TTG→CTG(Leu→Leu),Exon13 CD1436 TCT→TCC (Ser→Ser) ,and two kinds of missense mutation, which are Exon11 CD1038GAA→3GA (Glu→Gly) and Exon11 CD 1183 AAA→AGA(Lys→Arg).2. In 63 cases of breast cancer specimens, there are 32 missense mutation(CD1038 GAA→GGA (Glu1038Gly) ), 13 0f which most occurres in the invasive ductal carcinoma cases. Among 36 one level relatives cases having breast cancer risk, the case 26, 34, 110 are CD1038 GAA→GGA and the CD1183 AAA→AGA dual heterozygote carriers, whether to develop for breast cancer waits for tracing the revisit.3. MLPA testing did not find any major genome rearrangement events. After the examination of MLPA P002B BRCA1 reagent box and the analyses of Beckman 8000 GeneMapper, it discovered that 13 apparent child's suspicious flaws in one case by comparing with the control group. This result has prompt related with the BRCAl gene heterozygosity flaw. The MLPA P087 BRCAl reagent box verify the results did not find in exon 13 deletion and rearrangement of the situation.[ Conclusion ]1. This study detected four kinds of non-sense mutation, two kinds of missense mutation. This is a new mutation and never be reported. The result suggests the detection of BRCAl mutations is feasible in crowd and important to explore the pathogenesis of breast cancer.2. BRCA1 CD1038 GAA→GGA (Glu1038Gly) and CD1183 AAA→AGA(Lys1183Arg) mutation detected by high-frequency examineschanges, may be BRCA1 gene "the mutation hotspot" in Chinese crowd. Genetic family history of cases a marked increase in this risk, that such a mutation may occur in the breast ductal carcinoma.3. Direct PCR-DHPLC-sequencing and MLPA technology on the BRCAl gene mutation and re-scheduling is a convenient, fast, accurate and high rate of detection methods. However, the present study results suggest that MLPA technology BRCA1 gene rearrangements in our population of breast cancer-related events may be less. |
PDF Full Text Request