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

采用基因诱捕法建立Prxⅰ缺陷小鼠。将逆转录病毒基因诱捕载体插入Prx I位点的胚胎干细胞(Omni bank no.；OST422296)转移到养母身上产生嵌合动物（Lexicon Genetics Inc.）。通过回交将遗传背景统一为C57BL/6。将B16个小鼠黑色素瘤细胞（1 × 10^5个细胞/体）接种于三组（野生型、杂合型和纯合型Prxⅰ缺陷小鼠）小鼠足垫，观察淋巴结转移和肺转移。当遗传背景为C57/129Sv时，Prx I的丢失减少了淋巴结和肺转移。然而，经过5代回交和统一的C57BL/6遗传背景后，野生型和Prx I纯合缺陷小鼠的淋巴结和肺转移没有显著差异。当另一种细胞系（LLC细胞，来源于肺癌：3 × 10^6个细胞/体）接种于这些组时，没有差异。我们以Matrigel作为肿瘤生长基质，将B16细胞（1 × 105个细胞/体）与凝胶混合接种皮下，观察其血管形成情况，无显著差异。另一方面，我们研究了与Prx I受相同转录因子Nrf2调控的A170氧化应激的应激反应，分析氧化应激诱导氧化应激蛋白的机制。我们还研究了GATA-1转基因小鼠，因为Prx I和GATA-1都与红细胞异常有关。

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.