乳腺癌相关差异表达基因的筛选及分析

Screening and analysis of differentially expressed genes related to breast cancer

目的构建乳腺癌组织与癌旁正常乳腺组织差异表达的cDNA消减文库,筛选乳腺癌相关基因并进行分析。方法应用抑制性消减杂交(suppression subtractive hybridization,SSH)技术对15对乳腺癌组织及正常癌旁乳腺组织进行差异基因的消减,构建cDNA消减文库,并转化大肠杆菌进行文库扩增,采用数字表法随机挑选克隆经PCR鉴定后,对200个阳性克隆插入片段的序列进行测定及同源性分析。结果建立了乳腺癌差异表达片段cDNA消减文库;通过测序和同源性分析,得到173个差异表达基因,其中26个与肿瘤发生发展相关;26个基因中有15个已被文献明确报道在乳腺癌组织或细胞中有差异表达。结论应用SSH技术成功建立了乳腺癌差异表达基因cDNA消减文库,为乳腺癌的发病机制研究及寻找潜在的治疗靶点提供了有力的支持。

Objective To screen differentially expressed genes between breast cancer tissues and corresponding normal breast tissues by constructing a cDNA subtractive library with suppression subtractive hybridization （SSH） technique.Methods Fifteen pairs of breast cancer and corresponding normal breast tissues were studied to construct a cDNA subtractive library by SSH. Amplification of the library was carried out with E. coli strain DH5α. The cDNA was sequenced and analyzed with the relevant sequences in GenBank with Blast search after PCR. Results The subtracted cDNA library of differentially expressed genes of breast cancer was successfully constructed and 173 differentially expressed cDNA fragments were obtained. Homology analysis showed that 26 of them were tumor-associated genes in which 15 genes were reported to be related with breast cancer.Conclusion The construction of cDNA library of differentially expressed genes of breast cancer provides a useful pathway for exploring the pathogensis of breast cancer and looking for the potential target genes for treating breast cancer.