Analysis by cDNA microarrays of gene expression patterns of human adrenocortical tumors
Analysis by cDNA microarrays of gene expression patterns of human adrenocortical tumors
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DOI：
10.1530/eje.1.02116
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发表时间：
2006-04-01
影响因子：
5.8
通讯作者：
Bartsch, DK
中科院分区：
文献类型：
--
作者：
Slater, EP;Diehl, SM;Bartsch, DK
目的：肾上腺皮质癌（ACC）是一种罕见的恶性肿瘤，预后极差。肾上腺皮质肿瘤发生的分子机制仍不清楚。良性和恶性肾上腺皮质肿瘤的基因表达模式的cDNA微阵列的比较分析，使我们能够确定新的肿瘤抑制基因和原癌基因的基础adrenocortical tumorogenesis.Design and Methods：总RNA从新鲜冷冻组织的10 ACC和10良性肾上腺皮质腺瘤组织学确认后，至少85%的肿瘤细胞分离。参考由10个正常肾上腺皮质样本的合并RNA组成。扩增肿瘤和参考的RNA用于在翻转颜色技术中合成Cy 3-和Cy 5-荧光标记的cDNA。结果：ACC与正常组织、皮质腺瘤与正常组织、皮质腺瘤与皮质腺瘤之间差异表达4倍以上的基因分别为42个、11个、21个。如通过实时PCR证实的，IGF 2基因在ACC中与皮质腺瘤和正常皮质组织相比显著上调。在肾上腺皮质肿瘤中表达下调的基因包括嗜铬粒蛋白B和早期生长反应因子1。结论：应用基因芯片技术对肾上腺皮质肿瘤进行全面的基因表达谱分析，有助于阐明这些不明确肿瘤发生的分子机制。评估已鉴定基因的作用。需要进一步详细的分析，包括与临床数据的相关性。
Objectives: Adrenocortical carcinoma (ACC) is a rare malignant neoplasm with extremely poor prognosis. The molecular mechanisms of adrenocortical tumorigenesis are still not well understood. The comparative analysis by cDNA microarrays of gene-expression patterns of benign and malignant adrenocortical tumors allows us to identify new tumor-suppressor genes and proto-oncogenes underlying adrenocortical tumorigenesis.Design and methods: Total RNA from fresh-frozen tissue of 10 ACC and 10 benign adrenocortical adenomas was isolated after histologic confirmation of neoplastic cellularity of at least 85%. The reference consisted of pooled RNA of 10 normal adrenal cortex samples. Amplified RNA of tumor and reference was used to synthesize Cy3- and Cy5-fluorescently labeled cDNA in a flip-color technique. D-chips containing 11540 DNA spots were hybridized and scanned and the images were analyzed by ImaGene 3.0 software.Results: The comparative analysis of gene expression revealed many genes with more than fourfold expression difference between ACC and normal tissue (42 genes), cortical adenoma and normal tissue (11 genes), and ACC and cortical adenoma (21 genes) respectively. As confirmed by real-time PCR, the IGF2 gene was significantly upregulated in ACCs versus cortical adenomas and normal cortical tissue. Genes that were downregulated in adrenocortical tumors included chromogranin B and early growth response factor 1.Conclusions: Comprehensive expression profiling of adrenocortical tumors by the cDNA microarray technique is a very powerful tool to elucidate the molecular steps associated with the tumorigenesis of these ill-defined neoplasms. To evaluate the role of identified genes. further detailed analyses, including correlation with clinical data, are required.