Kobayashi Daiki, Hirayama Mio, Komohara Yoshihiro, Mizuguchi Souhei, Wilson Morifuji Masayo, Ihn Hironobu, Takeya Motohiro, Kuramochi Akira, Araki Norie
Department of Tumor Genetics and Biology, Kumamoto University, Kumamoto 860-8556, Japan.
Department of Cell Pathology, Graduate School of Medical Sciences, and Kumamoto University, Kumamoto 860-8556, Japan.
J Biol Chem. 2014 Sep 19;289(38):26314-26326. doi: 10.1074/jbc.M114.568253. Epub 2014 Aug 4.
Neurofibromatosis type 1 (NF1) is an autosomal dominant disease that predisposes individuals to develop benign neurofibromas and malignant peripheral nerve sheath tumors (MPNSTs). Due to the lack of information on the molecular mechanism of NF1-associated tumor pathogenesis or biomarkers/therapeutic targets, an effective treatment for NF1 tumors has not been established. In this study, the novel NF1-associated protein, translationally controlled tumor protein (TCTP), was identified by integrated proteomics and found to be up-regulated via activated MAPK/PI3K-AKT signaling in response to growth factors in NF1-deficient Schwann cells. Immunohistochemical analysis of NF1-associated tumors revealed that the TCTP expression level correlated with tumorigenicity. In NF1-deficient MPNST cells, TCTP protein but not mRNA was down-regulated by NF1 GTPase-activating protein-related domain or MAPK/PI3K inhibitors, and this correlated with suppression of mammalian target of rapamycin (mTOR) signaling. mTOR inhibition by rapamycin also down-regulated TCTP protein expression, whereas knockdown or overexpression of TCTP suppressed or activated mTOR signaling, respectively, and affected cell viability. These results suggest that a positive feedback loop between TCTP and mTOR contributes to NF1-associated tumor formation. Last, the anti-tumor effect of artesunate, which binds to and degrades TCTP, was evaluated. Artesunate significantly suppressed the viability of MPNST cells but not normal Schwann cells, and the TCTP level inversely correlated with artesunate sensitivity. Moreover, combinational use of artesunate and rapamycin enhanced the cytotoxic effect on MPNST cells. These findings suggest that TCTP is functionally implicated in the progression of NF1-associated tumors and could serve as a biological target for their therapy.
1型神经纤维瘤病(NF1)是一种常染色体显性疾病,使个体易患良性神经纤维瘤和恶性外周神经鞘瘤(MPNST)。由于缺乏关于NF1相关肿瘤发病机制的分子机制或生物标志物/治疗靶点的信息,尚未建立针对NF1肿瘤的有效治疗方法。在本研究中,通过整合蛋白质组学鉴定了新型NF1相关蛋白——翻译调控肿瘤蛋白(TCTP),发现在NF1缺陷的施万细胞中,TCTP通过激活的MAPK/PI3K-AKT信号通路对生长因子作出反应而上调。对NF1相关肿瘤的免疫组织化学分析显示,TCTP表达水平与致瘤性相关。在NF1缺陷的MPNST细胞中,NF1 GTP酶激活蛋白相关结构域或MAPK/PI3K抑制剂可下调TCTP蛋白而非mRNA,这与雷帕霉素靶蛋白(mTOR)信号通路的抑制相关。雷帕霉素抑制mTOR也下调TCTP蛋白表达,而敲低或过表达TCTP分别抑制或激活mTOR信号通路,并影响细胞活力。这些结果表明,TCTP与mTOR之间的正反馈环有助于NF1相关肿瘤的形成。最后,评估了与TCTP结合并降解TCTP的青蒿琥酯的抗肿瘤作用。青蒿琥酯显著抑制MPNST细胞的活力,但不影响正常施万细胞,TCTP水平与青蒿琥酯敏感性呈负相关。此外,青蒿琥酯与雷帕霉素联合使用增强了对MPNST细胞的细胞毒性作用。这些发现表明,TCTP在功能上与NF1相关肿瘤的进展有关,可作为其治疗的生物学靶点。