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IMPROVED DIAGNOSIS OF TUMORS IN COMPANION ANIMALS USING PCR METHOD

使用 PCR 方法改进伴侣动物肿瘤的诊断

基本信息

批准号：
08456163
负责人：
KUBO Kihei
金额：
$ 0.9万
依托单位：
OSAKA PREFECTURE UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08456163/
关键词：
erbB c-kit prohibitin marker chromosome RT-PCR mapping mammary tumor companion animal erb B 腺癌 erbB3 erbB4 クローニング肥満細胞腫 erb B1 erb B2

项目摘要

The expression of erbBl, B2, B3, B4 and c-kit proto-oncogene in canine and feline mammary tumor specimens was investigated to evaluate its potential usefulness as tumor markers. By comparing the homology among the nucleotide sequences of cDNA reported for several mammalian species, pairs of primers were synthesized for the reverse transcriptase-polymerase chain reaction (RT-PCR) method. The expression of c-kit transcript was detected in 11 mammary tumors (adenocarcinomas, benign and malignant mixed tumors). Of the 12 tumors originated from various tissues except mastocytomas, only low level of RT-PCR products iwre detected in 5 samples, whereas no apparent amplification was observed in 7 tumors. Any of erbB family was not expressed in normal mammary tissue, while the multiple expression was frequently observed in tumor samples. In three feline mammAry tumors, the frequent and multiple expression of erbB genes was also observed. Although c-kit transcript was detected in two samples, it was not apparent in one specimen in which the expressions of all of erbB genes were observed. These results indicate that the expression of erbB family and c-kit transcript is useful markers for the diagnosis of both canine and feline mammary tumors.The chromosome analysis of cell lines derived from feline mammary adenocarcinoma demonstrated that FMT-3 and FMT4 cells showed monosomy or complete alleic deletion of particular chromosomes which were included in specific marker chromosomes. In none of the feline tumors examined, RT-PCR product of prohibitin was detected. Feline prohibitin gene was mapped at the long arm of chromosome B4 by fluorescence in situ hybridization method. Because the karyotypes of both cell lines showed B4 trisomy, the chromosomal localization of the gene in FMT-3 cells was investigated. A couple of B4 in the cells was shown to have apparent doublet signals of hybridization.

为探讨erbB1、B2、B3、B4和c-kit原癌基因在犬和猫乳腺肿瘤标本中的表达，探讨其作为肿瘤标志物的可能性。通过比较已报道的几种哺乳动物的cDNA核苷酸序列的同源性，合成了一对用于逆转录聚合酶链式反应(RT-PCR)的引物。检测11例乳腺肿瘤(腺癌、良恶性混合瘤)中c-kit基因的表达。在除肥大细胞瘤外的12例肿瘤中，5例仅检测到低水平的RT-PCR产物，7例未见明显扩增。在正常乳腺组织中无一例表达，而在肿瘤组织中呈多发性表达。在三种猫乳腺肿瘤中，也观察到了erbB基因的频繁和多重表达。虽然在2例标本中检测到c-kit转录本，但在1例所有erbB基因表达的标本中检测不到c-kit转录本。这些结果表明，erb B家族和c-kit转录本的表达对犬和猫乳腺肿瘤的诊断都是有用的标记。对猫乳腺癌细胞系的染色体分析表明，FMT-3和FMT4细胞表现出特定的染色体单体或完全等位缺失，这些染色体包含在特定的标记染色体中。在所有被检测的猫科动物肿瘤中，都没有检测到禁止素的RT-PCR产物。利用荧光原位杂交技术，将猫科动物的禁忌素基因定位在染色体B4的长臂上。由于两株细胞的核型均为B4三体，因此对该基因在FMT-3细胞中的染色体定位进行了研究。细胞内有几个B4具有明显的双重杂交信号。