Identification of a novel tumor suppressor gene on chromosome arm 18q in human pancreatic caner

人胰腺癌染色体臂 18q 上新型抑癌基因的鉴定

基本信息

批准号：
18390118
负责人：
HORII Akira
金额：
$ 9.88万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390118/
关键词：
pancreatic cancer tumor suppressor gene 18q RNA interference miRNA 膵がん癌抑制遺伝子

项目摘要

Loss of 18q is highly frequent in human pancreatic cancer. A tumor suppressor gene, SMAD4, resides in this region, but introduction of this gene inhibit tumor cell growth only in vivo by means of angiogenesis inhibition through ETS. Introduction of chromosome 18, however, inhibit both in vitro and in vivo. Moreover, in most of the early lesions of the pancreatic tumorigeneses, 18q is lost but SMAD4 functions normally. Allelotype analyses on 18q identified a commonly lost region between D18S451 and D18S462. Database search indicated a total of 164 protein coding genes. One of such genes, PMAIP1, was picked up by the microarray analyses. This gene is frequently suppressed in pancreatic cancer. Upregulation of this gene caused suppression of the cell growth in pancreatic cancer cells. On the contrary, siRNA-mediated knockdown of the PMAIP1 stimulated cell growth. Thus, the PMAIP1 gene is one of the candidate tumor suppressor genes for pancreatic cancer.We further performed systematic knockdown studies using siRNAs for all the protein-coding genes in the commonly deleted region. Recovery of the cell growth was monitored and we picked up a total of 13 candidate genes. Further detailed studies are necessary.In the course of these studies, we developed a modified buffer system for ligation. Using this system, we can perform ligations in a cheap, efficient, and rapid manner. A total of 100 ligation reaction can be done within a 10 minutes only spending one dollar for 100 reactions. Thus, we named this method as "Coffee break ligation technique."

在人类胰腺癌中，18Q的损失高度频繁。肿瘤抑制基因SMAD4位于该区域，但是该基因的引入仅通过通过ETS抑制血管生成抑制体内抑制肿瘤细胞的生长。然而，引入18染色体，抑制了体外和体内。此外，在胰腺肿瘤的大多数早期病变中，18Q丢失，但SMAD4正常起作用。在18Q上进行的分析分析确定了D18S451和D18S462之间通常丢失的区域。数据库搜索表明总共有164个蛋白质编码基因。这样的基因之一PMAIP1被微阵列分析拾取。该基因经常在胰腺癌中受到抑制。该基因的上调导致胰腺癌细胞中细胞生长的抑制。相反，siRNA介导的PMAIP1敲低刺激了细胞生长。因此，PMAIP1基因是胰腺癌的候选肿瘤抑制基因之一。我们进一步使用siRNA进行了系统的敲低研究，用于常见删除区域中所有蛋白质编码基因。监测了细胞生长的恢复，我们总共获得了13个候选基因。在这些研究的过程中，我们开发了一种修改的缓冲系统进行连接。使用此系统，我们可以以便宜，高效且快速的方式进行连接。总共可以在10分钟内完成100次连接反应，仅花一美元才能进行100个反应。因此，我们将此方法命名为“咖啡断裂技术”。