Analysis of lymph node metastasis of oral cancer by using peroxiredoxin I knockout mouse

采用基因捕获法建立Prx I缺陷小鼠模型。将具有插入Prx I基因座的逆转录病毒基因捕获载体的胚胎干（ES）细胞（Omni bank no. OST 422296）转移至寄养母体以产生嵌合动物（Lexicon Genetics Inc.）。通过回交将遗传背景统一到C57 BL/6。将B16鼠黑色素瘤细胞（1 × 10^5个细胞/个体）接种到三组（野生型、杂合型和纯合型Prx I缺陷小鼠）的小鼠足垫中，观察淋巴结转移和肺转移。当基因背景为C57/129 Sv时，Prx I缺失可减少淋巴结和肺转移。然而，经过5代回交和统一的C57 BL/6遗传背景，野生型和Prx I纯合缺陷小鼠之间的淋巴结和肺转移没有显著差异。当在这些组中接种另一种细胞系（LLC细胞，来源于肺癌：3x10^6个细胞/个体）时，没有差异。通过使用Matrigel作为肿瘤生长的基质，我们将B16细胞（1 × 105个细胞/个体）和凝胶的混合物接种到皮下并研究血管化，但没有显著差异。另一方面，我们研究了A170细胞氧化应激的应激反应，该反应受与Prx I相同的转录因子Nrf 2调控，以分析氧化应激如何诱导氧化应激蛋白。由于Prx I和加塔-1都与红细胞异常有关，我们还对加塔-1转基因小鼠进行了研究。

Prx I deficient mouse was established by gene trapping method. Embryonic stem (ES) cells with a retroviral gene trap vector inserted into the Prx I locus (Omni bank no.OST422296) were transferred to a foster mother to generate chimeric animals (Lexicon Genetics Inc.). The genetic background was uniformized to C57BL/6 by back cross. B16 murine melanoma cells (1x10^5 cells/body) were inoculated into the mouse foot pad of three groups (wild-type, heterozygous, and homozygous Prx I deficient mouse) and lymph node metastasis and lung metastasis were observed. When genetic background was C57/129Sv, lymph node and lung metastasis were reduced by Prx I loss. However, after 5 generation back cross and uniformized C57BL/6 genetic background, there was no significant difference in lymph node and lung metastasis between wild-type and Prx I homozygous deficient mice. When another cell line (LLC cell, derived form lung cancer : 3x10^6 cells/body) was inoculated in these groups, there was no difference. By using Matrigel as a matrix of tumor growth, we inoculated the mixture of B16 cells (1x105 cells /body) and the gel into subcutaneous and investigated the vascularization, but there was no significant difference. On the other hand we investigated the stress response of A170 oxidative stress, which is regulated by same transcription factor Nrf2 as Prx I to analyze how oxidative stress protein was induce by oxidative stress. And we also investigated GATA-1 transgenic mouse, because Prx I and GATA-1 were both related with red blood cell abnormality.