Antitumor Effects of Proteasome Inhibition in Anaplastic Thyroid Carcinoma
Antitumor Effects of Proteasome Inhibition in Anaplastic Thyroid Carcinoma
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DOI：
10.2967/jnumed.111.101295
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发表时间：
2012-11-01
影响因子：
9.3
通讯作者：
Haberkorn, Uwe
中科院分区：
文献类型：
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作者：
Altmann, Annette;Markert, Annette;Haberkorn, Uwe
泛素-蛋白酶体通路已被确定为癌症治疗的潜在分子靶点。在这项研究中，我们研究了蛋白酶体抑制剂硼替佐米对甲状腺间变性癌（ATC）的作用，其特征是对多模式治疗方法完全无效。方法：将ATC细胞株0643和SW 1736用硼替佐米（1nM至1 μ M）处理12 - 72 h。此后，通过胸苷摄取实验和活细胞数测定分析生长抑制。测定细胞凋亡并进行细胞周期分析。利用基因芯片分析和实时定量PCR系统，我们测量了转录的变化。在荧光素酶活性测定中，使用报告构建体pNF-κ B-TA-Luc和pp53-TA-Luc监测核因子（NF）-κ B和p53信号转导途径的活性。使用H-3-FDG，C-14-氨基异丁酸和Na（125）碘化物进行摄取测量，以研究体外代谢变化和碘转运体活性。此外，F-18-FDG的摄取进行了评估，在ATC荷瘤裸鼠治疗后1或2天硼替佐米。结果如下：硼替佐米诱导SW 1736和C643细胞生长抑制、凋亡和G（2）-M细胞周期阻滞，并上调p21（CIP1/WAF1）表达。此外，葡萄糖代谢和氨基异丁酸摄取显着降低，在体外的ATC细胞系在体内只有在SW 1736肿瘤在2天后硼替佐米治疗。硼替佐米处理的SW1736和0643细胞的转录谱显示参与应激反应、凋亡、细胞周期调节和分化的基因表达增加。使用实时定量PCR定量基因表达，我们还注意到肿瘤坏死因子相关的骨化诱导配体和甲状腺特异性转录因子Pax8和TTF-1的上调，导致甲状腺特异性靶基因甲状腺球蛋白，钠碘同向转运体，甲状腺过氧化物酶和促甲状腺激素受体的表达，并在ATC细胞中适度积累碘。结论：硼替佐米是一种很有前途的治疗ATC的药物。为了改善临床结果，显然有必要进一步研究硼替佐米治疗甲状腺癌的潜力。
The ubiquitin-proteasome pathway has been identified as a potential molecular target for cancer therapy. In this study, we investigated the effect of the proteasome inhibitor bortezomib on anaplastic thyroid carcinoma (ATC) characterized by complete refractoriness to multimodal therapeutic approaches. Methods: The ATC cell lines 0643 and SW1736 were treated with bortezomib (1 nM to 1 mu M) for 12-72 h. Thereafter, growth inhibition was analyzed by thymidine uptake experiments and determination of the viable cell number. Apoptosis was measured and a cell cycle analysis was done. Using gene chip analysis and the real-time quantitative PCR system, we measured transcriptional changes. The activity of the nuclear factor (NF)-kappa B and p53 signal transduction pathways was monitored using the reporter constructs pNF-kappa B-TA-Luc and pp53-TA-Luc in the luciferase activity assay. Uptake measurements using H-3-FDG, C-14-aminoisobutyric acid, and Na(125)iodide were performed to investigate metabolic changes and iodide symporter activity in vitro. Moreover, the F-18-FDG uptake was evaluated in ATC tumor-bearing nude mice 1 or 2 d after treatment with bortezomib. Results: Bortezomib induced growth inhibition, apoptosis, and G(2)-M cell cycle arrest associated with upregulation of p21(CIP1/WAF1) expression in SW1736 and C643 cells. Moreover, the glucose metabolism and aminoisobutyric acid uptake significantly decreased in vitro in both of the ATC cell lines in vivo only in SW1736 tumors at 2 d after the bortezomib treatment. The transcriptional profile in bortezomib-treated SW1736 and 0643 cells revealed increased expression of genes involved in stress response, apoptosis, regulation of the cell cycle, and differentiation. Using real-time quantitative PCR for the quantification of gene expression, we additionally noticed upregulation of the tumor necrosis factor-related apoptosis-inducing ligand and the thyroid-specific transcription factors Pax8 and TTF-1, leading to expression of the thyroid-specific target genes thyroglobulin, sodium iodide symporter, thyroperoxidase, and thyroid-stimulating hormone receptor and to a moderate accumulation of iodide in ATC cells. Conclusion: On the basis of our data, bortezomib represents a promising antineoplastic agent for the treatment of ATC. To improve the clinical outcome, further investigation into the potential of bortezomib therapy of thyroid cancer is clearly warranted.