Jensen Kirk, Bikas Athanasios, Patel Aneeta, Kushchayeva Yevgeniya, Costello John, McDaniel Dennis, Burman Kenneth, Vasko Vasyl
Department of PediatricsUniformed Services University of the Health Sciences, Bethesda, Maryland, USA
Department of Internal MedicineGeorgetown University Hospital MedStar, Washington Hospital Center Internal Medicine Residency Program, Washington, District of Columbia, USA.
Endocr Relat Cancer. 2017 Mar;24(3):147-156. doi: 10.1530/ERC-16-0568. Epub 2017 Jan 30.
The HIV protease inhibitor Nelfinavir (NFV) inhibits PI3K/AKT and MAPK/ERK signaling pathways, emerging targets in thyroid cancers. We examined the effects of NFV on cancer cells that derived from follicular (FTC), papillary (PTC) and anaplastic (ATC) thyroid cancers. NFV (1-20 µM) was tested in FTC133, BCPAP and SW1736 cell lines. The effects of NFV on cell proliferation were determined using real-time microscopy and by flow cytometry. DNA damage, apoptotic cell death and expression of molecular markers of epithelial-mesenchymal transition (EMT) were determined by Western blot and real-time PCR. Real-time imaging demonstrated that NFV (10 µM) increased the time required for the cell passage through the phases of cell cycle and induced DNA fragmentation. Growth inhibitory effects of NFV were associated with the accumulation of cells in G0/G1 phase, downregulation of cyclin D1 and cyclin-dependent kinase 4 (CDK4). NFV also induced the expression of γH2AX and p53BP1 indicating DNA damage. Treatment with NFV (20 µM) resulted in caspase-3 cleavage in all examined cells. NFV (20 µM) decreased the levels of total and p-AKT in PTEN-deficient FTC133 cells. NFV had no significant effects on total ERK and p-ERK in BRAF-positive BCPAP and SW1736 cells. NFV had no effects on the expression of EMT markers (Twist, Vimentin, E- and N-Cadherin), but inhibited the migration and decreased the abilities of thyroid cancer cells to survive in non-adherent conditions. We conclude that NFV inhibits proliferation and induces DNA damage in thyroid cancer cell lines. Our data suggest that NFV has a potential to become a new thyroid cancer therapeutic agent.
艾滋病病毒蛋白酶抑制剂奈非那韦（NFV）可抑制PI3K/AKT和MAPK/ERK信号通路，而这两条信号通路是甲状腺癌中新出现的治疗靶点。我们研究了NFV对源自滤泡状甲状腺癌（FTC）、乳头状甲状腺癌（PTC）和间变性甲状腺癌（ATC）的癌细胞的影响。在FTC133、BCPAP和SW1736细胞系中测试了NFV（1 - 20 μM）。通过实时显微镜观察和流式细胞术确定NFV对细胞增殖的影响。通过蛋白质免疫印迹法和实时聚合酶链反应确定DNA损伤、凋亡性细胞死亡以及上皮-间质转化（EMT）分子标志物的表达。实时成像显示，NFV（10 μM）延长了细胞通过细胞周期各阶段所需的时间，并诱导了DNA片段化。NFV的生长抑制作用与细胞在G0/G1期的积累、细胞周期蛋白D1和细胞周期蛋白依赖性激酶4（CDK4）的下调有关。NFV还诱导了γH2AX和p53BP1的表达，表明存在DNA损伤。用NFV（20 μM）处理导致所有检测细胞中的半胱天冬酶-3裂解。NFV（20 μM）降低了PTEN缺陷的FTC133细胞中总AKT和磷酸化AKT的水平。NFV对BRAF阳性的BCPAP和SW1736细胞中的总ERK和磷酸化ERK没有显著影响。NFV对EMT标志物（Twist、波形蛋白、E-钙黏蛋白和N-钙黏蛋白）的表达没有影响，但抑制了迁移并降低了甲状腺癌细胞在非贴壁条件下的存活能力。我们得出结论，NFV抑制甲状腺癌细胞系的增殖并诱导DNA损伤。我们的数据表明，NFV有潜力成为一种新型的甲状腺癌治疗药物。
