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

1.在人纤维肉瘤细胞（HT 1080）转移中参与肿瘤-宿主基质细胞相互作用的基因的选择1）HT 1080的亲本和高转移性克隆之间的基因表达无显著差异从HT 1080选择性克隆高转移性克隆，并使用Human Cancer 1.2 Atlas Expression Arrays（Clonetech）通过cDNA宏阵列分析基因表达谱。HT 1080细胞株在体外培养的高转移克隆与其亲本细胞株之间的基因表达无明显差异。2）肺微转移中纤维连接蛋白1（FN 1）基因在HT 1080细胞中的高表达。在肺微转移中存在多个基因的过表达和低表达。其中，纤维连接蛋白1（FN 1）基因的表达比体外培养的表达增加了50倍。 ...更多信息 培养的HT 1080细胞。真实的时间RT-PCR证实FN 1基因转录显著上调，激光捕获显微切割/真实的时间RT-PCR证实FN 1基因过度表达定位于肺微转移的HT 1080肿瘤细胞。采用裸鼠皮下接种（H组）和肌肉接种（M组）模型，研究了HT 1080细胞侵袭转移过程中宿主间质细胞的相互作用。与H组相比，M组显示出显著增强的肿瘤细胞生长和肺微转移。使用人类癌症1.2 Atlas表达阵列和小鼠Atlas表达阵列分析肿瘤细胞以及M组和H组的宿主基质细胞的基因表达。阵列分析显示，与H组相比，M组连接斑珠蛋白基因表达显著下调。通过真实的时间RT-PCR和免疫组化方法分别在mRNA水平和蛋白水平证实连接斑珠蛋白基因表达下调。H-组和M-组间质细胞基因表达无明显差异.连接斑珠蛋白基因在软组织肉瘤中的表达：临床应用从蛋白和mRNA水平检测了连接斑珠蛋白基因在各种软组织肉瘤中的表达，结果显示，连接斑珠蛋白在滑膜肉瘤细胞中过表达，尤其在上皮型滑膜肉瘤细胞中。在恶性纤维组织细胞瘤（MFH）中，与无肺转移者相比，有肺转移者连接斑珠蛋白的表达无论在蛋白水平还是在mRNA水平均显著下调。少

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