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Analysis of the numerical aberration of chromosome 17 and 18 on frozen section of gastric adenocarcinomas

胃腺癌冰冻切片17、18号染色体数值畸变分析

基本信息

批准号：
07670196
负责人：
OOI Akishi
金额：
$ 1.47万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670196/
关键词：
Chromosome in situ hybridization Chromosome numerical aberration Gastric cancer DCC p53 FISH クロモゾーム17 クロモゾーム18 クロモソーム17 クロモソーム18

项目摘要

This study was designed to facilitate understanding of the process of gastric cancer progression by clarifying numerical changes of chromosomes 17 and 18 on gastric cancer sections. For DNA hybridization, we chose two chromosome-specific alpha-centromeric probes : one for chromosome 17 on whose short arm, tumor suppressor gene p53 located and the other for chromosome 18 on whose long arm tumor suppressor gene DCC located. In addition, to know the relationship between the numerical changes of these chromosomes and allelic loss of p53 and DCC,PCR-LOH was done using microsatelite polymorphism located at or linked to these genes.In order to dissolve technical problems, we first compared the results of hybridization done by using frozen sections, and by formalin-fixed paraffin-embedded sections. We confirmed that the hybridization was not less efficient when formalin-fixed paraffin-embedded tissues were used than when frozen sections were used. Second, we compared two detection methods, i.e … More ., fluorescence detection and peroxidase detection. By the former method, the specimen had disadvantage of fading of fluorescences, however because virtually no background staining, the enumeration was easier than when peroxidase detection was applied. Thus finally we chose fluorescence in situ hybridization (FISH) on formalin-fixed paraffin-embedded section as the regular detection tmethod.The 33 tumors which had allelic imbalance of p53 by PCR-LOH,and 20 tumors which had allelic imbalance of DCC were examined by FISH for numerical aberrations of chromosomes 17 and 18, respectively. FISH for chromosome 17 was successful in 28 tumors and for chromosome 18 in 18 tumors. Polysomy 17 was found in 14 and polysomy 18 in 7. No monosomy 17 was found, however monosomy 18 was found in 2 tumors. By PCR-LOH,the distinction of allelic imbalance due to numerical aberrations from those due to the selective physical deletion of genetic loci was impossible, although it is easy to detect numerical chromosomal aberrations by FISH.In this study it is shown that FISH is a useful tool to clarify chromosomal changes on formalin-fixed paraffin-embedded specimen. Less

本研究旨在通过阐明胃癌切片上17号和18号染色体的数量变化来促进对胃癌进展过程的了解。在DNA杂交中，我们选择了两个染色体特异的α着丝粒探针：一个位于17号染色体上，其短臂上有抑癌基因P53；另一个位于18号染色体上，其上有长臂抑癌基因DCC。此外，为了了解这些染色体的数目变化与P53和DCC等位基因丢失的关系，我们利用位于这些基因或与这些基因连锁的微卫星多态进行了PCR-LOH。为了解决技术问题，我们首先比较了冰冻切片和福尔马林固定石蜡切片的杂交结果。我们证实，当使用福尔马林固定的石蜡包埋组织时，杂交效率并不低于使用冰冻切片时的效率。其次，我们比较了两种检测方法，即…荧光检测和过氧化物酶检测。前一种方法有荧光褪色的缺点，但由于几乎没有背景染色，计数比用过氧化物酶检测更容易。因此，我们最终选择了福尔马林固定石蜡包埋切片上的荧光原位杂交(FISH)作为常规检测方法。对33例P53等位基因缺失的肿瘤和20例DCC等位基因失衡的肿瘤分别进行了17号和18号染色体的FISH检测。17号染色体FISH成功28例，18号染色体FISH成功18例。17号染色体多体型14例，18号染色体多体型7例。未发现17号单体，2例为18号单体。尽管FISH很容易检测到染色体数目异常，但通过聚合酶链式反应-杂合性缺失，无法区分由数目异常引起的等位基因失衡和由选择性物理缺失引起的等位基因失衡。较少