应用基因芯片技术研究甲状腺乳头状癌的基因表达

目的　应用基因芯片技术研究甲状腺乳头状癌 (PTC)和正常成人甲状腺组织基因的差异表达。方法　分别用Cy5和Cy3两种不同的荧光染料通过逆转录反应将PTC组和对照组甲状腺组织的mRNA分别标记成探针 ,并与载有一组靶基因的基因表达谱芯片进行杂交。通过扫描荧光强度 ,计算机软件分析 ,寻找两组差异表达基因 ,并用RT PCR、免疫组化对其中两条基因进行验证。结果　共有 65条差异表达基因 ,其中表达增加的有 48条 (2 .0倍以上 ) ,表达降低的有 17条 (0 .5倍以下 )。RT PCR、免疫组化结果与芯片扫描结果一致。结论　基因芯片是筛选PTC与正常成人甲状腺组织差异表达基因的有效方法。通过筛选所得差异基因提示 ,PTC的发病涉及细胞外基质、细胞因子、受体信号转导等多个方面。

Study on differentially expressed genes in papillary thyroid carcinoma by cDNA microarray

Objective To screen the differentially expressed genes between papillary thyroid carcinoma (PTC) and normal thyroid tissues using cDNA microarray. Methods mRNA from both PTC and normal thyroid tissues were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP (Cy-5 or Cy-3) to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray, which was scanned for the fluorescent signals and showed difference between two tissues. Two among the differentially expressed genes were verified by RT-PCR and immunohistochemistry. Results In PTC 48 genes were upregulated while 17 were downregulated. The results of RT-PCR and immunohistochemistry were consistent with that of the gene chips. Conclusion cDNA microarray technique is effective in screening the differentially expressed genes between 2 different kinds of tissue. The obtained genes are mainly related with extracellular matrix, cytokine, signal transduction and so on.