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

从人类染色体 3P 和 8P 克隆肿瘤抑制基因 (TSG)

• 批准号: 6289207
• 负责人: MICHAEL LERMAN
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6289207
• 关键词: chromosomes; clone cells; computer assisted sequence analysis; human genetic material tag; human tissue; lung neoplasms; molecular cloning; molecular oncology; neoplasm /cancer genetics; nucleic acid sequence; single strand conformation polymorphism; tumor suppressor genes

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 accomplishments this year are:(1) the novel pVHL target genes identified by us last year,namely,the carbonic anhydrases, CA9 and CA12 were further analyzed.We showed that the CA9 and CA12 genes are overexpressed in many tumor types due to the loss of VHL or other mechanisms and are involved in the control of the extracellular pH of the miliew surrounding the cells and thus create a microenvironment conducive to tumor growth and spread.Based on these finding Dr. E. Oldfield and a group of surgeons at the NIH Clinical Center initiated a prospective, non-randomized study of the effect of acetazolamide, a strong inhibitor of CAs, in patients with brain hemangioblastomas associated with brain tissue edema and cysts.Future work will focus on the role of carbonic anhydrases and othes genes in the regulation of tumor pH and its potential impact on cancer growth.(2) The 3p21.3 TSG:A subset of 19 genes found in the deletions overlap of 370-kb were subdivided by a nesting deletion into two gene sets: eight genes lying in the proximal 120-kb segment and 11 genes lying in the distal 250-kb segment. Both gene sets were analyzed extensively by manual and computational methods. Four of the 19 genes showed loss-of-expression or reduced mRNA levels in small cell (SEMA V) or non-small cell (a2d-2) or both (BLU and LUCA1) cancer cell lines. None of the 8 genes showed a frequent (>10%) mutation rate in lung cancer samples leading to conclude that with the exception of the three genes with reduced expression in this set they should be excluded as classical tumor suppressors in sporadic lung cancer. The mutation analysis of the 11 gene set is not yet completed and may reveal a TSG with homozygotic inactivation in tumors. Further mutational analysis in breast tumors and functional testing of the critical genes by gene transfer and gene disruption strategies is ongoing and should permit the identification of the putative TSG(s) among this gene set.(3) The 3p12 TSG:we discovered two new homozygous deletions in SCLC lines (250kb) removing part of the DUTT1 gene. We hypothesized that these deletions may target another cancer gene that may reside in a large intron of DUTT1. To search for this gene we prepared a sequence ready p1phage contig. We also requested the WU Sequencing Center to prepare a 1-mb BAC contig (between markers D3S3681 and D3S1604) that includes the deleted area for sequencing in the near future. (4) The 8p22 TSG:we established the intro-exon structure of the HP gene we cloned last year (GenBank # AFO26219) and are conducting mutation analyses in lung, breast, and prostate cancers. - Carcinogenesis, hereditary cancer, Lung cancer, Oncogenes, Tumor Suppressor, - Human Tissues, Fluids, Cells, etc.

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 accomplishments this year are:(1) the novel pVHL target genes identified by us last year,namely,the carbonic anhydrases, CA9 and CA12 were further我们表明，由于VHL或其他机制的丧失，CA9和CA12基因在许多肿瘤类型中过表达，并且参与了细胞周围的MILIEW的细胞外pH的控制，从而在这些发现的E. Oldfield和NIH的一组临床中启动的临床中心促进了肿瘤的生长，从而产生了一个微环境，从而创造了一个微观环境。与脑组织水肿和囊肿相关的脑血管血管群患者的乙酸唑胺是CAS的强抑制剂。FUTURE工作将集中于碳萎缩症和OTHES基因在调节肿瘤pH的调节中的作用及其对癌症生长的潜在影响。通过嵌套缺失成两个基因集：八个基因位于近端120-kb段中，远端250-kb段中的11个基因。通过手动和计算方法对两个基因组进行了广泛的分析。在这19个基因中，有4个表现出表达丧失或降低的小细胞（SEMA V）或非小细胞（A2D-2）或（BLU和LUCA1）癌细胞系中的mRNA水平降低。在肺癌样品中，没有8个基因在肺癌样品中频繁（> 10％）突变率得出结论，除了在此集合中降低表达的三个基因外，应将其排除在零星肺癌中的经典肿瘤抑制。 11个基因集的突变分析尚未完成，可能显示肿瘤中纯合灭活的TSG。通过基因转移和基因破坏策略对乳腺肿瘤进行进一步的突变分析以及对关键基因的功能测试正在进行中，应允许在此基因组中鉴定假定的TSG。（3）3P12 TSG：我们发现了SCLC线（250KB）中的两种新的型型型TSG。我们假设这些缺失可能针对另一个可能存在于DUTT1内含子的癌症基因。为了搜索此基因，我们准备了一个序列准备就绪的重叠群。我们还要求WU测序中心准备一个1-MB BAC重叠群（在标记D3S3681和D3S1604之间），其中包括在不久的将来进行测序的删除区域。 （4）8p22 TSG：我们于去年克隆的HP基因的介绍 - 介绍了 - ＃AFO26219），并正在肺，乳腺癌和前列腺癌进行突变分析。 - 致癌，遗传癌，肺癌，癌基因，肿瘤抑制因子，人体组织，液体，细胞等。