POSITIVE SELECTION OF CANDIDATE TUMOR-SUPPRESSOR GENES BY SUBTRACTIVE HYBRIDIZATION
POSITIVE SELECTION OF CANDIDATE TUMOR-SUPPRESSOR GENES BY SUBTRACTIVE HYBRIDIZATION
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DOI：
10.1073/pnas.88.7.2825
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发表时间：
1991-04-01
影响因子：
11.1
通讯作者：
SAGER, R
中科院分区：
文献类型：
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作者：
LEE, SW;TOMASETTO, C;SAGER, R
描述了一种设计用于识别和恢复候选肿瘤抑制基因的正选择系统。该系统比较了在一种特殊培养基中生长的正常和肿瘤来源的人乳腺上皮细胞的基因mRNA表达，这种培养基支持两种细胞类型的相似生长速度。在正常细胞中唯一表达的mRNA与肿瘤细胞的mRNA相减后恢复为cdna。在分析的前23个克隆中恢复了7个不同的克隆，转录本大小从0.6到4.8千碱基不等，包括稀有和丰富的转录本。在分离的克隆中，有编码缝隙连接蛋白连接蛋白26、两种不同的角蛋白和谷胱甘肽- s -转移酶pi的基因，以及小钙结合蛋白S100家族中的一个未知基因。原则上，肿瘤抑制基因包括两类：一类是由于DNA突变或缺失导致功能丧失的基因；二类是由于调控阻滞导致功能丧失的基因。II类抑制基因被认为是由另一种因突变或缺失而失去功能的抑制基因调控的。这两类肿瘤抑制基因可能为癌症诊断或治疗提供有价值的蛋白质。II类抑制因子可能特别有用，因为正常基因存在，它们的重新表达可能被药物或其他治疗诱导。
A positive selection system designed to identify and recover candidate tumor-suppressor genes is described. The system compares mRNA expression of genes from normal and tumor-derived human mammary epithelial cells grown in a special medium that supports similar growth rates of the two cell types. mRNAs uniquely expressed in normal cells are recovered as cDNAs after subtraction with mRNA from tumor cells. Seven different clones, from 0.6 to 4.8 kilobases in transcript size and including both rare and abundant transcripts, were recovered in the first 23 clones analyzed. Among the isolated clones were genes encoding the gap-junction protein connexin 26, two different keratins, and glutathione-S-transferase-pi, as well as an unknown gene in the S100 family of small calcium-binding proteins. In principle, tumor-suppressor genes include two classes: class I, in which loss of function results from mutation or deletion of DNA and class II, in which loss of function is from a regulatory block to expression. A class II suppressor gene is assumed to be regulated by a different suppressor gene that lost its function by mutation or deletion. Both classes of tumor-suppressor genes may provide valuable proteins with clinical applications in cancer diagnosis or therapy. Class II suppressors may be especially useful because the normal genes are present and their reexpression may be inducible by drugs or other treatments.