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目的检测 BRCA1和 BRCA2基因在散发性乳腺癌中的突变情况,探讨 BRCA1和BRCA2基因突变与乳腺癌的关系。方法选取2000年12月至2005年9月收治的144例乳腺癌患者标本作实验组,另取非癌乳腺组织标本30例作对照组。用酚-氯仿抽提法提取 DNA。针对各个外显子的碱基序列特征,设计有助于筛查基因碱基突变的聚合酶链反应(PCR)引物。每例 DNA 标本均用 PCR 扩增 BRCA1基因的全部22个外显子和 BRCA2基因的 exon10和exon14两个外显子。分别将每例外显子的 PCR 扩增产物进行单链构象多态性分析,对泳动变位或出现异常区带的 PCR 扩增产物进行 DNA 测序。结果对照组未检测出 BRCA1和 BRCA2基因突变,实验组中检测出20例 BRCA1基因碱基改变,总突变率为13.9%,其中错义突变率为11.1%。BRCA2基因 exon10和 exonl4未检测出突变。结论 BRCA1突变与乳腺癌密切相关,筛查 BRCA1基因突变对于中国人群乳腺癌患病风险评估、早期诊断及基因治疗具有重要意义。
Objective To detect the mutations of BRCA1 and BRCA2 in sporadic breast cancer and to explore the relationship between BRCA1 and BRCA2 mutations and breast cancer. Methods A total of 144 breast cancer patients from December 2000 to September 2005 were selected as experimental group and 30 non-cancer breast tissue samples as control group. Extraction of DNA with phenol - chloroform extraction. Based on the nucleotide sequence characteristics of each exon, polymerase chain reaction (PCR) primers were designed to screen gene mutations. In each DNA sample, all 22 exons of the BRCA1 gene and two exons of exon 10 and exon 14 of the BRCA2 gene were amplified by PCR. The PCR amplification products of each exon were analyzed by single-strand conformation polymorphism, and the DNAs of the PCR amplification products were analyzed by electrophoresis. Results The BRCA1 and BRCA2 gene mutations were not detected in the control group. Twenty cases of BRCA1 gene mutations were detected in the experimental group, with a total mutation rate of 13.9% and a missense mutation rate of 11.1%. No mutation was detected in exon10 and exonl4 of BRCA2 gene. Conclusion The mutation of BRCA1 is closely related to breast cancer. Screening for BRCA1 gene mutation is of great significance for the risk assessment, early diagnosis and gene therapy of breast cancer in Chinese population.