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胃癌的间期细胞遗传学:应用于从福尔马林固定石蜡包埋组织中获取细胞的荧光原位杂交(FISH)

Interphase cytogenetics of gastric carcinoma: fluorescence in situ hybridization (FISH) applied to cells obtained from formalin-fixed paraffin-embedded tissues.

作者信息

Gomyo Y, Andachi H, Nagao K, Ikeguchi M, Ito H

机构信息

First Department of Pathology, Faculty of Medicine, Tottori University, Yonago, Japan.

出版信息

Pathol Int. 1995 Mar;45(3):227-32. doi: 10.1111/j.1440-1827.1995.tb03446.x.

Abstract

The interphase cytogenetics in formalin-fixed and paraffin-embedded gastric cancer tissues were examined by fluorescence in situ hybridization (FISH) with alpha-satellite DNA probes. Two gastric carcinoma cell lines, TMK-1 and MKN-28, were first analyzed cytogenetically. Of 25 TMK-1 cell karyotypes, chromosome 7 showed trisomy and chromosome 17 showed disomy in 18 cells. Most MKN-28 cells showed disomy of both chromosomes 7 and 17. Suspensions of singly isolated TMK-1 and MKN-7 cells were obtained from the cultured cells, and from paraffin-embedded tissue specimens fixed with formalin for 0, 1, 3 and 5 days obtained from xenotransplanted tumors in nude mice. The numbers of chromosomes 7 and 17 analyzed with the karyotypic preparations coincided well with those determined by FISH, even in the paraffin-embedded specimens. The number of tumor cells showing no signals, however, increased in the specimens after 5 days formalin fixation. In 10 surgically removed gastric carcinomas, the predominant signal number for chromosomes 7 and 17 in the cells of paraffin-embedded tissues was two (disomy), except in one papillary carcinoma, which was trisomic for chromosome 7. Large subpopulations (more than 20%) showing trisomy were found in four cases for chromosome 7 and in five cases for chromosome 17. A higher frequency of trisomy was found in well differentiated than in poorly differentiated carcinomas. These findings suggest that the FISH technique is a useful tool for detecting chromosomal aberrations in gastric adenocarcinoma cells, even in paraffin-embedded specimens, as long as the tissues are fixed with formalin for an appropriate time.

摘要

采用α-卫星DNA探针,通过荧光原位杂交(FISH)技术检测福尔马林固定石蜡包埋胃癌组织中的间期细胞遗传学。首先对两种胃癌细胞系TMK-1和MKN-28进行细胞遗传学分析。在25个TMK-1细胞核型中,18个细胞的7号染色体呈三体性,17号染色体呈二体性。大多数MKN-28细胞的7号和17号染色体均呈二体性。从培养细胞以及从裸鼠体内异种移植瘤获取的福尔马林固定0、1、3和5天的石蜡包埋组织标本中,获得单个分离的TMK-1和MKN-7细胞悬液。即使在石蜡包埋标本中,通过核型分析制备检测的7号和17号染色体数量与FISH检测结果也非常吻合。然而,福尔马林固定5天后的标本中,未显示信号的肿瘤细胞数量增加。在10例手术切除的胃癌中,石蜡包埋组织细胞中7号和17号染色体的主要信号数为两个(二体性),但有一例乳头状癌的7号染色体为三体性。在4例7号染色体和5例17号染色体中发现了显示三体性的大亚群(超过20%)。高分化癌中三体性的频率高于低分化癌。这些发现表明,只要组织用福尔马林固定适当时间,FISH技术就是检测胃腺癌细胞染色体畸变的有用工具,即使在石蜡包埋标本中也是如此。

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