Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference.
Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference.
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发表时间：
2003-03
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通讯作者：
N. Williams;R. Gaynor;S. Scoggin;U. Verma;T. Gokaslan;C. Simmang;J. Fleming;Denise Tavana;E. Fren
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作者：
N. Williams;R. Gaynor;S. Scoggin;U. Verma;T. Gokaslan;C. Simmang;J. Fleming;Denise Tavana;E. Fren
本研究的目的是分析结直肠肿瘤中基因表达的变化，以确定治疗这种疾病的新靶点和策略。实验设计cDNA微阵列分析用于检测正常组织和分离自20名患者的结肠肿瘤和息肉之间的基因表达差异。为了鉴定在调节结直肠癌生长特性中重要的基因，使用RNA干扰（RNAi）来破坏结肠肿瘤细胞系HCT116中几种过表达基因的表达，该细胞系显示出与许多患者肿瘤相似的基因表达模式。结果在大约三分之一的患者中，在总共9592个基因中的2632个（574个上调基因和2058个下调基因）中始终观察到表达变化≥ 2倍。随后通过实时定量PCR对13个基因的分析证实了该分析的可靠性。RNAi介导的这些基因之一，生存素，一种有效的细胞凋亡抑制剂的表达的破坏，严重降低了肿瘤生长在体外和体内异种移植模型。结论微阵列分析和RNAi技术的联合应用提供了一个很好的系统来确定在癌症中上调的特定基因的作用，这些基因导致结肠肿瘤的体外和体内生长增加。
PURPOSE The purpose of this study was to profile gene expression changes in colorectal tumors to identify new targets and strategies for the management of this disease. EXPERIMENTAL DESIGN cDNA microarray analysis was used to detect differences in gene expression between normal tissue and colon tumors and polyps isolated from 20 patients. To identify genes that are important in regulating the growth properties of colorectal cancer, RNA interference (RNAi) was used to disrupt expression of several of the overexpressed genes in a colon tumor cell line, HCT116, which showed similar patterns of gene expression as many of the patient tumors. RESULTS Expression changes of > or =2-fold in approximately one-third of the patients were consistently observed for 2632 of a total of 9592 genes (574 up-regulated genes and 2058 down-regulated genes). Subsequent analysis of 13 genes by quantitative real-time PCR confirmed the reliability of this analysis. RNAi-mediated disruption of the expression of one of these genes, survivin, a potent inhibitor of apoptosis, severely reduced tumor growth both in vitro and in an in vivo xenograft model. CONCLUSIONS The combined use of microarray analysis and RNAi provides an excellent system to define the role of specific genes that are up-regulated in cancer lead to the increased in vitro and in vivo growth of colon tumors.