Research on the Factor that Determines Susceptibility to Chemotherapy for Musculoskeletal Sarcoma by Genomic/Proteomic Analysis

本研究旨在利用基因组/蛋白质组学分析方法，阐明影响骨肉瘤、尤文氏肉瘤和胃肠道间质瘤(GIST)等原发肌肉骨骼肉瘤的预后因素和治疗易感性因素。这项研究的一个特点是通过使用实际的临床肌肉骨骼肉瘤样本，详尽而完整地阐明决定预后和治疗敏感性的基因和蛋白表达。从各种恶性软组织肉瘤和胃肠道间质瘤的冰冻标本中提取RNA/蛋白质，用Affymetrix U133基因芯片进行穷尽基因表达分析，用荧光二次电泳质谱进行穷尽蛋白质表达分析。基于基因芯片对恶性软组织肉瘤基因表达的详尽分析，我们发现所谓的恶性纤维组织细胞瘤可分为几个亚型…这进一步表明，根据基因表达模式，有可能分别分化为平滑肌肉瘤、脂肪肉瘤等。我们还通过穷举蛋白表达分析确定了恶性纤维组织细胞瘤不同组织学类型的蛋白质，并报道原肌球蛋白亚型的表达参与了平滑肌肉瘤亚型的亚型划分。通过对GIST进行详尽的蛋白表达分析，我们发现一种蛋白的表达水平在术后一年内早期复发者的手臂(预后较差)和无病存活两年以上的手臂(预后良好)之间存在显著差异。在基因芯片分析中，编码该蛋白的mRNA在其表达水平上也显示了类似的变化。这种蛋白被称为pfetin，在GIST手术标本的抗pfetin抗体免疫染色和Western blotting中，预后良好和预后不良的患者之间也存在显著差异(p&lt；0.0001)。我们还进行了原发肌肉骨骼肉瘤的平行临床和临床病理研究。较少

This research is aimed to clarify prognostic factors and factors related to the treatment susceptibility of primary musculoskeletal sarcoma including osteosarcoma, Ewing's sarcoma and gastrointestinal stromal tumor (GIST) using genomic/proteomic analytical methods. A feature of the research was to clarify gene and protein expression that determined the prognosis and susceptibility to treatment exhaustively and integratively by using actual clinical musculoskeletal sarcoma samples. RNA/protein were extracted from frozen samples of various malignant soft tissue sarcomas and GISTs, and exhaustive gene expression analyses were carried out using Affymetrix U133 GeneChip and exhaustive protein expression analyses using fluorescence secondary electrophoresis/mass spectrometry. Based on the results of the exhaustive gene expression analyses of malignant soft tissue sarcomas using GeneChip, we found that so-called malignant fibrous histiocytoma (MFH) could be subclassified into several subtypes … More that suggested a tendency to respectively differentiate into leiomyosarcoma, liposarcoma, etc. depending on the gene expression pattern. We also identified proteins that featured respective histological types of malignant fibrous histiocytoma by exhaustive protein expression analysis, and reported that the expression of tropomyosin subtype was involved in the subclassification of leiomyosarcoma into its subtypes. As a result of exhaustive protein expression analyses of GISTs, we detected a protein that was significantly different in its expression levels between the arm of early relapsers within one year after surgery (poor prognosis), and the arm of disease-free survivals for over two years (good prognosis). The mRNA that encoded this protein demonstrated similar changes in its expression levels also in the GeneChip analysis. This protein is called pfetin, and a significant difference (p<0.0001) was also found between the good prognosis and the poor prognosis arms in the antipfetin antibody immunostaining and Western blotting using GIST surgery samples. We also conducted parallel clinical and clinicopathologic research of primary musculoskeletal sarcoma. Less