Department of Internal Medicine, University of Pisa, School of Medicine, Via Roma 67, I-56100, Pisa, Italy.
J Clin Endocrinol Metab. 2011 Feb;96(2):E288-96. doi: 10.1210/jc.2010-1905. Epub 2010 Dec 8.
We have studied the antitumoral activity of two new pyrazolo[3,4-d]pyrimidine compounds (CLM3 and CLM29) in primary papillary dedifferentiated thyroid cancer (DePTC) cells.
The antiproliferative effect was tested in DePTC cells obtained at reoperation from patients with recurrence of the tumor. The concentrations of CLM3 and CLM29 used in the in vitro experiments were 1, 10, 30, and 50 μm.
Proliferation assays in DePTC cells showed a significant reduction of proliferation by CLM3 and CLM29, which was by 12% with CLM3 (the most potent compound) 10 μm, 43% with CLM3 30 μm, and 60% with CLM3 50 μm. CLM3 and CLM29 increased the percentage of apoptotic cells in DePTC cells dose dependently (P < 0.001) and inhibited migration (P < 0.001). A DePTC cell line (AL) was injected sc in CD nu/nu mice, and tumor masses became detectable 10 d after xenotransplantation. CLM3 (40 mg/kg · die) significantly inhibited tumor growth and weight, and the therapeutic effect was significant starting on the 19th day after cell implantation (4 d after the beginning of treatment). The CLM3-treated group of animals did not show any appreciable toxicity. CLM3 and CLM29 increased thrombospondin-1 expression in the AL cell line. A significant reduction of microvessels and in the percentage of antivascular endothelial growth factor antibody immunoreactivity was observed in the CLM3 treated tumors, with a simultaneous increase of the percentage of necrosis.
The antitumoral activity of two new pyrazolo[3,4-d]pyrimidine compounds (CLM3, CLM29) in vitro and CLM3 in vivo in DePTC has been shown, opening the way to a future clinical evaluation.
我们研究了两种新的吡唑并[3,4-d]嘧啶化合物（CLM3 和 CLM29）在原发性甲状腺乳头状去分化癌（DePTC）细胞中的抗肿瘤活性。
在肿瘤复发的患者再次手术时获得的 DePTC 细胞中测试了抗增殖作用。体外实验中使用的 CLM3 和 CLM29 浓度分别为 1、10、30 和 50μm。
DePTC 细胞增殖试验显示 CLM3 和 CLM29 显著抑制了细胞增殖，CLM3（最有效化合物）10μm 时抑制率为 12%，30μm 时为 43%，50μm 时为 60%。CLM3 和 CLM29 剂量依赖性地增加 DePTC 细胞中的凋亡细胞百分比（P < 0.001）并抑制迁移（P < 0.001）。将 DePTC 细胞系（AL）sc 注射到 CD nu/nu 小鼠中，异种移植后 10d 可检测到肿瘤肿块。CLM3（40mg/kg·天）显著抑制肿瘤生长和重量，从细胞植入后第 19 天（治疗开始后第 4 天）开始治疗效果显著。接受 CLM3 治疗的动物组未表现出任何明显的毒性。CLM3 和 CLM29 增加了 AL 细胞系中血小板反应蛋白-1 的表达。在 CLM3 治疗的肿瘤中观察到微血管和血管内皮生长因子抗体免疫反应性百分比的显著减少，同时坏死百分比增加。
两种新的吡唑并[3,4-d]嘧啶化合物（CLM3、CLM29）在体外和 CLM3 在体内对 DePTC 的抗肿瘤活性已被证明，为未来的临床评估开辟了道路。
