Validation of met as a therapeutic target in alveolar and embryonal rhabdomyosarcoma
Validation of met as a therapeutic target in alveolar and embryonal rhabdomyosarcoma
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DOI：
10.1158/0008-5472.can-05-4292
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发表时间：
2006-05-01
期刊：
影响因子：
11.2
通讯作者：
Ponzetto, Carola
中科院分区：
文献类型：
--
作者：
Taulli, Riccardo;Scuoppo, Claudio;Ponzetto, Carola
横纹肌肉瘤（RIMS）是一种高度恶性的儿童软组织肿瘤，来源于骨骼肌细胞。RMS可分为两种主要的组织学亚型：胚胎型（ERMS）和肺泡型（ARMS），后者的特征在于PAX3/7-FKHR易位。在这里，我们首先研究了Met受体，PAX3和PAX7的转录靶点，是否在PAX3-FKHR介导的转化中起作用。在PAX3-FKHR转导后，Met在小鼠胚胎成纤维细胞（MEF）、NIH 3T3和C2C12细胞中表达上调，并且它们都获得了锚定独立性。这种特性在低血清中丧失，但加入肝细胞生长因子/分散因子（HGF/SF）挽救了软琼脂生长。通过用来源于Met突变体（Meto（D/D））和野生型（Met（+/+））胚胎的PAX3-FKHR MEF转导，获得Met对于这种PAX3-FKHR介导的效应是必需的遗传证据。只有Met（+/+）MEFs获得了锚定非依赖性生长，而PAX3-FKHR转导的Met（D/D）细胞不能在软琼脂中形成集落。为了验证Met是否在RMS维持中发挥作用，我们通过用表达抗Met短发夹RNA（shRNA）的诱导型慢病毒转导ERMS和ARMS细胞系来沉默受体。Met下调显著影响RIMS细胞的增殖、存活、侵袭力和非贴壁依赖性生长。最后，Met指导的shRNA的诱导促进了RMS异种移植模型中肿瘤质量的显著减少。我们的数据表明，ARMS和ERMS衍生的细胞系，尽管可能已经发生了多年的文化的遗传漂移，似乎保留了“成瘾”的Met癌基因，并建议Met可能代表一个目标的选择，开发新的治疗策略ARMS。
Rhabdomyosarcoma (RIMS) is a highly malignant soft-tissue tumor of childhood deriving from skeletal muscle cells. RMS can be classified in two major histologic subtypes: embryonal (ERMS) and alveolar (ARMS), the latter being characterized by the PAX3/7-FKHR translocation. Here we first investigated whether the Met receptor, a transcriptional target of PAX3 and PAX7, has a role in PAX3-FKHR-mediated transformation. Following PAX3-FKHR transduction, Met was up-regulated in mouse embryonal fibroblasts (MEF), NIH 3T3 and C2C12 cells, and they all acquired anchorage independence. This property was lost in low serum but addition of hepatocyte growth factor/scatter factor (HGF/SF) rescued soft-agar growth. Genetic proof that Met is necessary for this PAX3-FKHR-mediated effect was obtained by transducing with PAX3-FKHR MEFs derived from Met mutant (Meto(D/D)) and wild-type (Met(+/+)) embryos. Only Met(+/+) MEFs acquired anchorage-independent growth whereas PAX3-FKHR-transduced Met(D/D) cells were unable to form colonies in soft agar. To verify if Met had a role in RMS maintenance, we silenced the receptor by transducing ERMS and ARMS cell lines with an inducible lentivirus expressing an anti-Met short hairpin RNA (shRNA). Met down-regulation significantly affected RIMS cells proliferation, survival, invasiveness, and anchorage-independent growth. Finally, induction of the Met-directed shRNA promoted a dramatic reduction of tumor mass in a xenograft model of RMS. Our data show that both ARMS- and ERMS-derived cell lines, in spite of the genetic drift which may have occurred in years of culture, seem to have retained an "addiction" to the Met oncogene and suggest that Met may represent a target of choice to develop novel therapeutic strategies for ARMS.