Prognostic implication of MET overexpression in myxofibrosarcomas: an integrative array comparative genomic hybridization, real-time quantitative PCR, immunoblotting, and immunohistochemical analysis
Prognostic implication of MET overexpression in myxofibrosarcomas: an integrative array comparative genomic hybridization, real-time quantitative PCR, immunoblotting, and immunohistochemical analysis
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DOI：
10.1038/modpathol.2010.128
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发表时间：
2010-10-01
期刊：
影响因子：
7.5
通讯作者：
Huang, Hsuan-Ying
中科院分区：
文献类型：
--
作者：
Lee, Jen-Chieh;Li, Chien-Feng;Huang, Hsuan-Ying
粘液纤维肉瘤的发生、发展和转移的基础仍不清楚。7号染色体的增加在一些肉瘤中很常见，包括粘液纤维肉瘤，而具体的癌基因还有待鉴定。我们对7q31.2的MET基因进行了整合研究，以阐明其在粘液纤维肉瘤中的意义。以7号染色体上的候选癌基因为研究对象，采用385 K微阵列比较基因组杂交技术，对12个样本的DNA拷贝数变化进行了分析。实时定量RT-PCR成功定量MET转录16激光显微切割肿瘤和两个粘液纤维肉瘤细胞系（NMFH-1，OH 931）。MET免疫表达在随访的86例原发性局部肿瘤中可评估。为了分析内源性MET表达和激活，对均具有野生型MET基因的NMFH-1和OH 931细胞进行蛋白质印迹，并评估肝细胞生长因子处理的NMFH-1细胞的MET酪氨酸磷酸化动力学。在5例病例中检测到7 q上的非随机大规模增益，描绘了3个复发扩增子，7q21.11-7q21.3，7q22.1-22.3和7q31.1-7q32.3，其中MET基因座显示拷贝数增加等。MET mRNA在OH 931、NMFH-1和9个肿瘤（56%）中上调，而MET的基因剂量和mRNA表达均与临床病理因素无关。相比之下，MET蛋白过度表达，存在于67%的病例中，与深部位置（P=0.004）、较高级别（P=0.001）和较晚期（P
It remains obscure in myxofibrosarcoma about the basis of tumorigenesis, progression, and metastasis. Chromosome 7 gains are common in some sarcomas, including myxofibrosarcoma, whereas the specific oncogenes are yet to be characterized. We performed an integrative study of MET gene at 7q31.2 to elucidate its implication in myxofibrosarcoma. Focused on candidate oncogenes on chromosome 7, 385K array comparative genomic hybridization was used to profile DNA copy number alterations of 12 samples. MET transcript was successfully quantified by real-time RT-PCR for 16 laser-microdissected tumors and two myxofibrosarcoma cell lines (NMFH-1, OH931). MET immunoexpression was assessable in 86 primary localized tumors with follow-up. To analyze endogenous MET expression and activation, NMFH-1 and OH931 cells, both with wild-type MET gene, were subjected to Western blotting and hepatocyte growth factor-treated NMFH-1 cells were evaluated for the kinetics of MET tyrosine phosphorylation. Non-random large-scale gains on 7q were detected in five cases, delineating three recurrent amplicons, 7q21.11-7q21.3, 7q22.1-22.3, and 7q31.1-7q32.3, in which the locus of MET displayed increased copy number, among others. MET mRNA was upregulated in OH931, NMFH-1, and nine tumors (56%), whereas neither gene dosage nor mRNA expression of MET was associated with clinicopathological factors. In contrast, MET protein overexpression, present in 67% of cases, was highly related to deep location (P=0.004), higher grades (P=0.001), and more advanced stages (P