Expression profiling of genes involved in tumor - host stroma cell interaction in the metastasis of human fibrosarcoma cell (HT1080)

人纤维肉瘤细胞（HT1080）转移中涉及肿瘤-宿主基质细胞相互作用的基因的表达谱

• 批准号: 15590322
• 负责人: UEDA Yoshimichi
• 金额: $ 1.47万
• 依托单位: KANAZAWA MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590322/
• 关键词: human soft tissue sarcoma; HT1080; metastasis; cDNA macroarrays; fibronectin 1 gene; junction plakoglobin gene; pulmonary micrometastasis; orthotopic inoculation; ヒト線維肉腫細胞; 宿主・間質相互応答; cDNAマイクロアレイ; Plakoglobin遺伝子; 肺転移; 悪性線維性組織球腫; 滑膜肉腫; c-DNA

1. Selection of genes involved in the tumor-host stroma cell interaction in the metastasis of human fibrosarcoma cell (HT1080)1) No significant difference of gene expressions between parent and highly metastatic clones of HT1080Highly metastatic clones were selectively cloned from HT1080 and gene expression profilings were analysed by cDNA macroarrays using Human Cancer 1.2 Atlas Expression Arrays (Clonetech). No significant difference of gene expressions between parent and highly metastatic clones of HT1080 in vitro cultures was found out.2) Fibronectin 1 (FN 1) gene is overexpressed in HT1080 cells in pulmonary micrometastasesGene expressions involved in pulmonary micrometastasis-formation were investigated with a lung metastatic model of HT1080 using nude mice injected through tail vein. Several overexpressed and underexpressed genes were demonstrated in pulmonary micrometastasis. Of them, expression of fibronectin 1 (FN 1) gene was increased 50 times compared with that of in vitro- … More cultured HT1080 cells. The prominent upregulation of FN 1 gene transcription was verified by real time RT-PCR and the overexpression was localized in embolized HT1080 tumor cells of the pulmonary micrometastases by laser captured microdissection / real time RT-PCR.3) Junction plakoglobin gene is down-regulated in orthotopically inoculated intramuscle tumor cells showing enhanced growth and pulmonary micrometastasesGenes involved in the tumor - host stroma cell interaction in the invasion & metastasis of HT1080 cells were investigated using heal pad inoculation (H-group) and intramuscular inoculation (M-group) models in nude mice. M-group showed significantly enhanced tumor cell growth and pulmonary micro-metastases compared with H-group. Gene expressions of tumor cells as well as host stroma cells of M- and H-groups were profiled using both Human Cancer 1.2 Atlas Expression Arrays and Mouse Atlas Expression Arrays. Array-analyses disclosed that expression of junction plakoglobin gene was significantly down-regulated in M-group compared with that of H-group. The down-regulation of junction plakoglobin gene was confirmed by real time RT-PCR at mRNA level and by immunohistochemistry using specific monoclonal antibody to plakoglobin at protein level. No significant genes differently expressed between stroma cells of H- and M-groups were shown.2. Involvement of the down regulation of junction plakoglobin in human soft tissue sarcomas : Clinical applicationExpressions of junction plakoglobin gene were evaluated at both protein and mRNA levels in various kinds of human soft tissue sarcomas.Junction plakoglobin was overexpressed in synovial sarcoma cells, especially epithelial type. In malignant fibrous histiocytomas (MFH), expressions of junction plakoglobin was significantly down-regulated, at not only protein but also mRNA level, in tumors with pulmonary metastases compared with those without pulmonary metastases. Less

1。在人类纤维肉瘤细胞转移中涉及的基因选择（HT1080）1）在HT1080和Gene表达中选择了HT1080高转移性克隆的父母和高度转移性转移性克隆之间的基因表达和高度转移性转移性克隆的基因表达差异。 （Clonetech）。在HT1080在体外培养物中，父母和高度转移性克隆之间的基因表达没有显着差异。2）纤连蛋白1（Fn 1）基因在HT1080细胞中通过使用HTASTASTATIC MAGETATIC模型的肺部微构症调查了肺微含量模型的HT1080细胞中的HT1080细胞中的基因（FN 1）基因。在肺动脉灾脉中证明了几种过表达和未被压制的基因。其中，与体外培养的HT1080细胞相比，纤连蛋白1（FN 1）基因的表达增加了50倍。 The prominent update of FN 1 gene transcription was verified by real time RT-PCR and the overexpression was localized in embolized HT1080 tumor cells of the pulmonary micrometastases by laser captured microdissection / real time RT-PCR.3) Junction plakoglobin gene is down-regulated in orthotopically inoculated intramuscle tumor cells showing enhanced growth and pulmonary micrometastasesGenes involved in使用HEAL PAD接种（H组）和肌肉内接种（M-GROUP）模型，研究了HT1080细胞侵袭和转移中的肿瘤 - 基质细胞相互作用。与H组相比，M群显示出明显增强的肿瘤细胞生长和肺微发生酶。肿瘤细胞的基因表达以及M-组和H组的宿主基质细胞使用人类癌症1.2 Atlas表达阵列和小鼠Atlas表达阵列进行了分析。 Array-Analyses透露，与H组相比，在M组中，连接plakoglobin基因的表达显着下调。连接plakoglobin基因的下调通过实时RT-PCR在mRNA水平和免疫组织化学上使用特定的单克隆抗体在蛋白质水平上证实。在H组和M群的基质细胞之间未表达任何明显的基因。2。连接plakoglobin在人软组织肉瘤中的下降调节的参与：在各种人类软组织肉瘤中，在蛋白质和mRNA水平上均评估了连接plakoglobin基因的临床应用表达。Jununctionplakoglobin在合基肉瘤细胞中过表达plakoglobin，尤其是上皮类型。在恶性纤维组织细胞瘤（MFH）中，与没有肺转移的肿瘤相比，在具有肺转移的肿瘤中，连接plakoglobin的表达不仅在蛋白质和mRNA水平下显着下调。