CLONING TUMOR SUPPRESSOR GENES (TSG) FROM HUMAN CHROMOSOMES 3P AND 8P

这项研究的目标是识别、克隆和表征TSG 位于染色体3p和8p上，涉及起源或 人类主要恶性肿瘤的发展：肺癌， 乳房、肾脏和前列腺。我们今年取得的主要成就是： (1)VHL基因全序列组装(GenBank #AFO10238)；(2)差异鉴定pVHL靶基因 显示技术；到目前为止，25%的细胞转基因基因是 分析并鉴定了6个下调基因，即Noch 2， 指定细胞命运决定，机架1是细胞内受体 PKC、CA9和10(碳酸酐酶超家族成员)和两个 新的未知基因。(3)VHL启动子甲基化分析 肾癌：单染色体基因转移、细胞融合和VHL 基因转染法检测结果显示，表型以meth+为主。(4) 3p21.3TSG：去年定义并测序的370kb关键区域为 现在由于新发现的重叠缺失而缩小到120kb。 新的基因存在于该区域，包括一个新的α2-增量亚单位 诱导自身抗体的L型钙通道的表达 与小细胞肺癌相关的LEM(Lambert-Eaton肌萎缩症)。这些 正在分析基因的突变、表达、印记和 抑制靶癌细胞生长的能力。(5)3p12 TSG：A 候选基因是细胞黏附分子的新成员 超家族(CAMS)与TSG DCC的同源性正在分析中 (6)3p25-p26 TSG：用于电子定位克隆新成员 CALL被分离并分析(GenBank#AFO02246)。(7)8p22 TSG： NF1和TSC2这两个经典TSG的产物都是大GTP酶激活的 Ras和RAP1的蛋白质(GAP)。为了发现其他GAP，我们 在NCBI数据库中搜索代表TSGS区域这些基因的EST 由染色体8P和1上的LOH定义。我们现在已经分离出和 表征了大鼠p122RhoGAP的人类同源基因，这是一个双重的 RHO和PLC Delta的调节器。该基因被我们定位于8p22。 (GenBank#AFO26219)，是肺、乳房和 前列腺癌。

The goal of this research is to identify,clone,and characterize TSGs located on chromosomes 3p and 8p that are involved in the origin or development of major human malignancies: carcinomas of the lung, breast,kidney, and prostate. Our major accomplishments this year are: (1) assembly of the complete sequence of the VHL gene (GenBank #AFO10238);(2) identification of pVHL target genes by differential display technology; to date 25% of the cell transcibed genes were analyzed and six down regulated genes identified, namely,NOTCH 2,that specifies cell fate determination,RACK 1 an intracellular receptor for PKC, CA9 and10 (members of the carbonic anhydrase superfamily), and two new unknown genes. (3) analysis of the methylation of the VHL promoter in renal carcinoma:monochromosome gene transfer, cell fusion, and VHL gene transfections showed that the meth+ phenotype is dominant. (4)The 3p21.3 TSG:the 370kb critical region defined and sequenced last year is now narrowed to 120kb by a newly discovered overlapping deletion.Eight new genes reside in this region,including a new alpha2- delta subunit of the L-type calcium channel that appears to elicit autoantibodies in LEMS (Lambert - Eaton myastenic syndrom) associated with SCLC. These genes are being analyzed for mutations, expression,imprinting, and the ability to suppress growth of target cancer cells. (5)The 3p12 TSG: a candidate gene that is a new member of the cell adhesion molecule superfamily (CAMs) and showed homology to the TSG DCC is being analyzed (6)The 3p25-p26 TSG:using in silico location cloning a new member of CAMs,CALL,was isolated and analysed (GenBank #AFO02246).(7)The 8p22 TSG: products of two classical TSGs, NF1 and TSC2 are large GTPase activating proteins (GAPs) for RAS and RAP1 respectively.To discover other GAPs we searched NCBI databases for ESTs representing such genes in TSGs regions defined by LOH on chromosomes 8p and 1p.We have now isolated and characterized the human ortholog of the rat p122RhoGAP, which is a dual regulator of Rho and PLC delta. This gene is localized by us to 8p22 (GenBank # AFO26219) and is a candidate TSG for lung, breast, and prostate cancers.