基因芯片在宫颈癌、卵巢癌和乳腺癌相关基因表达研究中的运用

目的 应用基因微矩阵芯片筛查妇科恶性肿瘤相关基因表达。方法 分别提取癌组织和正常对照组织mRNA，分别用Cv3—dCTP和Cy5—dCTP经反转录荧光标记cDNA，获得两组探针，将探针混合后与基因芯片H40s(基因点数为4096点，上海联合基因公司提供)杂交，经严格洗片后用GenePix4000B扫描仪进行扫描，获得荧光信号图像，GenePixPro3．0图像处理软件对图像进行处理，获得两种组织中差异表达的基因信息。结果 在子宫颈癌组织中差异表达的基因24．61％(1008／4096)，其中表达降低(下调趋势)占11．28％(462／4096)；表达增高(上调趋势)占13．32％(546／4096)。在卵巢癌组织中差异表达的基因24．02％(984／4096)，其中表达降低(下调趋势)占12．67％(519／4096)，表达增高(上调趋势)占11．35％(465／4096)；在乳腺癌组织中差异表达的基因占9．67％(396／4096)，其中表达降低(下调趋势)占4．59％(188／4096)，表达增高(上调趋势)的5．08％(208／4096)。在3种癌组织中同时出现表达差异的基因有1．37％。结论 子宫颈癌、卵巢癌和乳腺癌有多种基因表达失衡。

Application of the cDNA microarray in analysis of gene expression patterns in cervical carcinoma, ovarin carcinoma and breast carcinoma

Purpose To screen the differentially expressed genes in malignant tumor of gynaecology by cDNA microarray. Methods The mRNA was isolated and purified from carcinoma and normal tissues and then were reversely transcribed to cDNAs with the incorporation of fluorescent Cy3 dCTP and Cy5 dCTP respectively to prepare two groups of hybridization probes.The mixed probes were hybridized to the cDNA microarray H40s( 4 096 gene points;provided by United Gene Holding Ltd,Shanghai). After high stringent washing,the cDNA microarray was scanned for the fluorescent signals by the scanner GenePix4000B,then the fluorescent signals were treated with image manipulation software GenePix Pro3 0 to show differentially expressed genes between the two tissues. Result Among the 4 096 target genes in cervical squamous carcinoma tissues, 24 61%genes( 1 008 / 4 096 ) expressed differently, 11 28% (462/ 4 096 ) of whom were downregulated, 13 32%(546/ 4 096 ) of whom were upregulated. Among the 4 096 target genes in ovarian malignant clear cell carcinoma tissues, 24 02%genes(984/ 4 098 ) expressed differently, 12 67%(519/ 4 096 ) of whom were downregulated, 11 35%(465/ 4 096 ) of whom were upregulated. In breast duct soakage carcinoma tissues, 9 67%(396/ 4 096 )genes expressed differently, 4 59%(188/ 4 096 )of whom were downregulated, 5 08%(208/ 4 096 )of whom were upregulated, 1 37% genes were much different in expression level in three kinds of carcinoma tissues. Conclusion The expression of many kinds of genes in cervical squamocellular carcinoma, ovarian malignant clear cell carcinoma and breast carcinoma tissues is altered.