Onoda Naoyoshi, Nakamura Masanori, Aomatsu Naoki, Noda Satoru, Kashiwagi Shinichiro, Kurata Kento, Uchino Shinya, Hirakawa Kosei
Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Osaka 545-8585, Japan.
Noguchi Thyroid Clinic, Beppu, Oita 874-0932, Japan.
Mol Clin Oncol. 2015 May;3(3):522-526. doi: 10.3892/mco.2015.496. Epub 2015 Jan 23.
We previously demonstrated the efficacy of gefitinib, a tyrosine kinase inhibitor of the epidermal growth factor receptor (EGFR), on an anaplastic thyroid cancer (ATC) cell line. We also observed that gefitinib was not effective in regulating cell growth in a different ATC cell line that exhibited an altered EGFR-initiated signal transduction pathway. In the present study, we attempted to regulate the downstream effector of EGFR-Akt-mammalian target of rapamycin (mTOR) pathway by an mTOR inhibitor, everolimus. A total of 8 ATC cell lines were employed, 7 of which were established in our institute. OCUT-2 was known to carry a mutation in the phosphoinositide-3-kinase, catalytic, α polypeptide gene (PI3KCA) and to be gefitinib-resistant, whereas ACT-1 exhibited a remarkable growth arrest by gefitinib. All the cell lines were tested for the cytotoxic effect of everolimus. The mechanisms of cellular toxicity were investigated by EGFR stimulation, cell cycle and concurrent exposure to paclitaxel. In OCUT-2, but not in any of the other cell lines, everolimus achieved a significant growth inhibition (inhibition of 30 and 50% was achieved by concentrations of 0.8 and 5 nM, respectively). The growth in OCUT-2 was inhibited by everolimus, even with concordant EGFR stimulation. This effect was demonstrated by a G2M cell cycle arrest. An additive effect of everolimus onto the cytotoxic effect of paclitaxel was demonstrated at a dose of 1-2 nM. A significant growth inhibitory effect of everolimus on the gefitinib-resistant ATC cell line was demonstrated, suggesting a possible correlation between the efficacy of everolimus and PI3KCA gene mutation and the significance of molecular-targeted therapy in the management of ATC.
我们之前已证明表皮生长因子受体(EGFR)的酪氨酸激酶抑制剂吉非替尼对一种间变性甲状腺癌(ATC)细胞系具有疗效。我们还观察到,吉非替尼在调节另一种显示EGFR启动的信号转导途径改变的ATC细胞系中的细胞生长方面无效。在本研究中,我们试图通过一种mTOR抑制剂依维莫司来调节EGFR-Akt-雷帕霉素哺乳动物靶点(mTOR)途径的下游效应器。总共使用了8种ATC细胞系,其中7种是在我们研究所建立的。已知OCUT-2在磷脂酰肌醇-3-激酶催化α多肽基因(PI3KCA)中携带突变且对吉非替尼耐药,而ACT-1对吉非替尼表现出显著的生长停滞。对所有细胞系进行了依维莫司的细胞毒性测试。通过EGFR刺激、细胞周期以及与紫杉醇同时暴露来研究细胞毒性机制。在OCUT-2中,而非其他任何细胞系中,依维莫司实现了显著的生长抑制(分别通过0.8 nM和5 nM的浓度实现了30%和50%的抑制)。即使存在一致的EGFR刺激,依维莫司也能抑制OCUT-2中的生长。这种效应通过G2M期细胞周期停滞得以证明。在1 - 2 nM的剂量下,依维莫司对紫杉醇的细胞毒性具有相加作用。证明了依维莫司对吉非替尼耐药的ATC细胞系具有显著的生长抑制作用,提示依维莫司的疗效与PI3KCA基因突变之间可能存在关联,以及分子靶向治疗在ATC管理中的重要性。