brca2基因启动子区多态性在乳腺癌预后中的分析及brca2基因启动子的克隆和研究word格式论文.docxVIP

BRCA2 基因启动子区多态性在乳腺癌预后的研究及BRCA2 基因启动子的克隆和分析英文摘要ANALYSIS OF PROMOTER PLOYMORPHISM OF BRCA2 IN ASSOCIATION WITH BREAST CANCER PROGNOSIS AND CLONING AND ANALYSIS OF THE BRCA2 PROMOTERAbstractPart1. Analysis of promoter polymorphism of BRCA2 in association with breast cancer prognosisObjective: To analysis the functional polymorphism in the promoter region of the BRCA2 gene with breast cancer prognosis.Methods: We performed study in 491 breast cancer cases to test the association between four polymorphisms and breast cancer prognosis.Genotypes were determined in DNA from blood cells by direct sequencing.Results: The result showed that -1144C/T polymorphism was associated with clinical outcome. Carriers of the TT genotype had a longer disease free survival (p=0.029). The result was more evident among sporadic breast cancer patients with unilateral cancer (p=0.010). Linkage disequilibrium (LD) analysis showed that all the five SNPs are in Linkage disequilibrium (D’0.8).Conclusions: This study showed that the -1144C/T polymorphism in BRCA2 gene promoter may affect prognosis of breast cancer in Chinese. Its significance in other populations remains to be investigated.英文摘要BRCA2 基因启动子区多态性在乳腺癌预后的研究及BRCA2 基因启动子的克隆和分析Part2:Cloning and analysis of the BRCA2 promoterObjective: To construct a BRCA2 promotor luciferase reporter gene vector and to analyze the mechanism of the gene regulation.Methods: By using genomic DNA from healthy people an a template, the BRCA2 promotor region was amplified by high fidelity DNA polymerase. Then the fragment was cloned into a recombinant plasmid. To analyze whether polymorphism rs3092989(-254A/G) could affect promoter activity, PCR-based mutagenesis was performed to generate haplotype containing -254G allele. Affer confirmed by direct sequencing, the BRCA2 promotor was subclonged into pGL3-basic vector. The constructed vector was transfected into plasmid HeLa cells to detect promoter activity.Results: Direct sequencing and restriction endonuclease