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Analysis of molecular mechanisms of cancer metastasis using transgenic mice

利用转基因小鼠分析癌症转移的分子机制

基本信息

批准号：
09470062
负责人：
TAKAHASHI Masahide
金额：
$ 8.51万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470062/
关键词：
transgenic mouse metastasis RET gene MMP TIMP medullary thyroid carcinoma malignant melanoma MEN2A チロシンキナーゼ ret遺伝子 MMP-9

项目摘要

We established a metallothionein-I (MT)/RET transgenic mouse line in which skin melanosis, benign melanocytic tumor and malignant melanoma develop stepwise. Malignant melanoma cells but not benign melanocytic tumor cells had metastatic ability in transgenic mice. In this study, we investigated the expression of several matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) including MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MT1-MMP, TIMP-1 and TIMP-2 in these tumors. Western and northern blot analyses revealed that malignant transformation of melanocytic tumors developed in MT/RET transgenic mice accompanied upregulation of MMP-9 and downregulation of TIMP-2. Expression of other MMP and TIMP genes examined was very low or undetectable in both benign and malignant tumors. Since activation of MMP-9 in malignant tumors was shown by gelatin zymography, these results suggest that imbalance of expression of the MMP-9 and TIMP-2 genes might be associated with metastatic ability of melanoma cells developed in MT/RET transgenic mice.In addition, we generated transgenic mice by introducing the RET-MEN2A gene fused to Moloney murine leukemia virus long terminal repeat. Expression of the transgene and its product was detected at variable levels in a variety of tissues including thyroid, heart, liver, colon, parotid gland and brain. All of 29 mice analyzed developed thyroid C cell hyperplasia or medullary carcinoma, accompanying high levels of serum calcitonim. Moreover, development of mammary or parotid gland adenocarcinoma was observed in half of the transgenic mice. Mammary carcinoma but not thyroid carcinoma often metastasized to lung. Thus, this transgenic mouse line could provide a useful system to analyze the genes responsible for metastasis.

我们建立了金属硫蛋白-I（MT）/RET转基因小鼠系，其中皮肤黑变病、良性黑素细胞肿瘤和恶性黑素瘤逐步发展。在转基因小鼠中，恶性黑色素瘤细胞具有转移能力，而良性黑色素细胞肿瘤细胞则没有。在这项研究中，我们研究了几种基质金属蛋白酶（MMP）和基质金属蛋白酶组织抑制剂（TIMP）在这些肿瘤中的表达，包括MMP-1、MMP-2、MMP-3、MMP-7、MMP-9、MT1-MMP、TIMP-1和TIMP-2。 Western和Northern印迹分析显示，MT/RET转基因小鼠中发生的黑素细胞肿瘤的恶性转化伴随着MMP-9的上调和TIMP-2的下调。所检查的其他 MMP 和 TIMP 基因的表达在良性和恶性肿瘤中都非常低或检测不到。由于明胶酶谱显示恶性肿瘤中 MMP-9 的激活，这些结果表明 MMP-9 和 TIMP-2 基因表达失衡可能与 MT/RET 转基因小鼠中发育的黑色素瘤细胞的转移能力有关。此外，我们通过引入与莫洛尼鼠白血病病毒长末端重复融合的 RET-MEN2A 基因来产生转基因小鼠。在多种组织中检测到转基因及其产物的表达水平不同，包括甲状腺、心脏、肝脏、结肠、腮腺和大脑。所分析的 29 只小鼠全部出现甲状腺 C 细胞增生或髓样癌，并伴有高水平的血清降钙素。此外，在一半的转基因小鼠中观察到出现乳腺癌或腮腺腺癌。乳腺癌（而非甲状腺癌）经常转移至肺部。因此，这种转基因小鼠系可以提供一个有用的系统来分析负责转移的基因。