应用基因芯片分析妇科恶性肿瘤的基因表达

目的应用基因微矩阵芯片研究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,经反转录分别用Cy3-dCTP和Cy5-dCTP荧光标记cDNA,获得两组探针,混合后与基因芯片H40s(基因点数为4096点)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,用图像处理软件:GenePix Pro3.0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4种癌组织中同时出现表达差异的基因有35条,占0.85%.其中4种癌组织中都下调表达的基因6条,没有均上调表达的基因,在前3种癌组织中都上调表达而在乳腺癌中却下调表达的基因12条,在前3种癌组织中都下调表达而在乳腺癌中却上调表达的基因12条,其它5条.结论 1.肿瘤的发生过程比我们已了解的要复杂的多,它是多种基因表达失衡的结果.每种肿瘤的每个患病个体其肿瘤的发生和发展都具有自己一系列独特的基因表达谱,即使是同一肿瘤的不同个体之间,基因表达谱可能差异也很大.2.子宫颈癌、乳腺癌、卵巢癌和子宫内膜癌这四种癌组织中具有相同表达行为的基因只有6个,它们是:AB023136(编码人类蛋白 KIAA0919)、AF080158(丝氨酸、苏氨酸K激酶基因)、S79522(蛋白质合成延长因子EF-1基因)、NM_000984(核糖体蛋白基因)、M32110(细胞增殖核抗原蛋白P120基因)、NM_000978(核糖体蛋白RPL23)基因,并且均下调表达.3. 子宫颈癌、子宫内膜癌和卵巢癌基因表达谱的一致性较高,而与乳腺癌基因表达谱的差异很大,说明3种内生殖器肿瘤发生的机理与乳腺癌的发病机理可能完全不同,有12个基因在子宫颈癌、子宫内膜癌和卵巢癌中上调表达而在乳腺癌中下调表达;同时另外12个基因在子宫颈癌、子宫内膜癌和卵巢癌中下调表达而在乳腺癌中上调表达.4.基因芯片是筛查妇科恶性肿瘤相关基因表达的有力工具.

Application of the cDNA microarry in Analysis of gene expression patterns in malignant tumor of gynaecology

Objective: To screen the differentially expressed genes in malignant tumor of gynaecology by cDNA microarry. Methods: The mRNA were isolated and purified from carcinoma and normal tissues then were reversely transcribed to cDNAs with the incorporation of fluorescent Cy3-dCTP or Cy5-dCTP one from the other to prepare two groups of hybridization probes , the mixed probes were hybridized to the cDNA microarray H40s(4096 gene points). After high-stringent washing, the cDNA microarry 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 each 2 tissues. Result: 35 of 4096(0.85%) genes were much different in expression level in four kinds of carcinoma tissues. 6 of 4096 were downregulated in four kinds of carcinoma, none of 4096 were upregulated genes,12 genes of 4096 were upregulated in cervical squamocellular carcinoma, ovarian malignant clear cell carcinoma and endometrial adenocarcinoma in uterus,while they were downregulated in breast duct soakage carcinoma.12 genes of 4096 were downregulated in cervical squamocellular carcinoma, ovarian malignant clear cell carcinoma and endometrial adenocarcinoma in uterus,while 5 of 4096 were upregulated in breast duct soakage carcinoma. Conclusion: 1.The origination and development of cancer is much more complicated than what we have known,it is the result to which the balance of many genes' expression were broken. The gene expression pattern is much different between various tumours or even same kind of tumour from different patients. 2.The similar expression pattern of 6 genes was observed in cervical squamocellular carcinoma, ovarian malignant clear cell carcinoma, endometrial adenocarcinoma in uterus and breast duct soakage carcinoma,they are : AB023136(Homo sapiens mRNA for KIAA0919 protein); AF080158 (Protein-Serine-Threonine Kinases); S79522 (The human gene sequences encoding the translation-associated functions of alpha-subunit of elongation factor 1 (EF-1 alpha) and the ubiquitin carboxyl extension protein (HUBCEP80). They have significantly higher levels of expression in fibroadenomas (benign) compared with carcinomas (malignant) of the breast.); NM_000984(Ribosomal Proteins); M32110 (The 120 kDa proliferating-cell nucleolar antigen,it is the most cancer specific of the proliferation-associated nucleolar proteins identified thus far.); NM_000978(ribosomal protein L17). 3.The gene expression pattern in cervical squamocellular carcinoma,ovarian malignant clear cell carcinoma and endometrial adenocarcinoma in uterus is identical in some degree,but is very various with that in breast duct soakage carcinoma,which indicates that it is possible for the pathogenesis in 3 kinds inner genitalia tumours is completely dissimilarity with that in breast duct soakage carcinoma,it is because that there are 12 genes in cervical squamocellular carcinoma,ovarian malignant clear cell carcinoma , endometrial adenocarcinoma in uterus were upregulated ,in opposition to downregulating in breast duct soakage carcinoma; on the other hand,others 12 genes in three kinds inner genitalia tumours downregulated ,upregulated in breast duct soakage carcinoma. 4.The cDNA microarray technique is a effective tool in screening the differentially expressed genes between 2 different kinds of tissue in malignant tumors of gynaecology.