5′-AMP-Activated Protein Kinase Regulates Papillary (TPC-1 and BCPAP) Thyroid Cancer Cell Survival, Migration, Invasion, and Epithelial-to-Mesenchymal Transition
5′-AMP-Activated Protein Kinase Regulates Papillary (TPC-1 and BCPAP) Thyroid Cancer Cell Survival, Migration, Invasion, and Epithelial-to-Mesenchymal Transition
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DOI：
10.1089/thy.2015.0440
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发表时间：
2016-07-01
期刊：
影响因子：
6.6
通讯作者：
Carvalho, Denise P.
中科院分区：
文献类型：
--
作者：
Cazarin, Juliana M.;Coelho, Raquel G.;Carvalho, Denise P.
背景：分化型甲状腺癌（DTC）预后良好，生存率高。然而，大约20-30%的DTC患者发生肿瘤复发，这加强了寻找新的分子靶点对癌症治疗的重要性。研究表明，5′-腺苷酸活化蛋白激酶（AMPK）在乳头状甲状腺癌（PTC）中过度活化。本研究旨在探讨AMPK激活剂5-氨基咪唑-4-carboxamide-核糖核苷（AICAR）对人甲状腺癌细胞BCPAP和TPC-1中细胞存活、凋亡、迁移、侵袭和上皮-间质转化（EMT）等各方面行为的影响。方法：在Dulbecco改良Eagle培养基中培养BCPAP和TPC-1细胞，在RPMI中培养非肿瘤源性细胞系nhy - ori。细胞在不同的时间内接受或不接受AICAR处理。利用transwell插入物分析细胞生长速率、细胞周期、细胞凋亡、细胞迁移和侵袭，并通过间充质和上皮标记物的表达来量化EMT。结果：AMPK在甲状腺癌细胞中被激活，AICAR治疗进一步增加了AMPK的磷酸化。AICAR处理48h后，两种细胞系G2/M期细胞比例下降，均出现G0/ g1期停滞。AMPK激活能有效诱导BCPAP和TPC-1癌细胞凋亡，而在nty - ori细胞中未观察到凋亡诱导作用。AICAR还使Nthy-ORI和BCPAP细胞在TPC-1细胞中的迁移分别减少30%和约60%。AICAR对Nthy-ORI和TPC-1细胞的细胞侵袭无影响，但对BCPAP细胞的细胞侵袭有显著降低。AICAR诱导BCPAP细胞N-cadherin显著降低，而vimentin和TCF/Zeb1蛋白的表达没有变化。在aicar处理的Nthy-ORI细胞中，EMT标记物的表达没有差异。在TPC-1细胞中检测到vimentin、TCF/Zeb1和N-cadherin蛋白表达显著降低。结论：通过抑制细胞增殖、细胞迁移和诱导细胞死亡，PTC细胞系中AMPK激活的增加导致了强烈的抗肿瘤反应。AMPK激活也逆转了TPC-1细胞中的EMT。
Background: Differentiated thyroid carcinomas (DTC) are associated with a good prognosis and a high survival rate. However, tumor recurrence occurs in approximately 20-30% of DTC patients, reinforcing the importance of identifying new molecular targets for cancer management. It has been shown that the 5 '-AMP-activated protein kinase (AMPK) is over-activated in papillary thyroid cancer (PTC). This study aimed to investigate the effects of 5-aminoimidazole-4-carboxamide-ribonucleoside (AICAR), an AMPK activator, on various aspects of thyroid cancer cell behavior, including cell survival, apoptosis, migration, invasion, and epithelial-to-mesenchymal transition (EMT), in the human thyroid cancer cell lines BCPAP and TPC-1.Methods: BCPAP and TPC-1 cells were cultivated in Dulbecco's modified Eagle's medium, and the non-tumor-derived cell line Nthy-ORI was grown in RPMI. Cells were treated or not with AICAR for different periods of time. The cell growth rate, cell cycle phase, apoptosis, cell migration, and invasion were analyzed using transwell inserts, and EMT was quantified by the expression of mesenchymal and epithelial markers.Results: AMPK is activated in thyroid cancer cell lines, and AICAR treatment further increased AMPK phosphorylation. After 48 hours of AICAR treatment, the percentage of cells in the G2/M phase decreased, and a G0/G1-phase arrest was induced in both cell lines. AMPK activation effectively induced apoptosis in the BCPAP and TPC-1 cancer cell lines, while no apoptosis induction was observed in Nthy-ORI cells. AICAR also reduced the migration of Nthy-ORI and BCPAP cells by 30% and approximately 60% in TPC-1 cells. AICAR had no effect on cell invasion in Nthy-ORI and TPC-1 cells, but a significant reduction of cell invasion was observed in BCPAP cells. AICAR induced a significant reduction of N-cadherin and no changes in the expression of vimentin or TCF/Zeb1 protein in BCPAP cells. No differences in the expression of EMT markers were found in the AICAR-treated Nthy-ORI cells. A remarkable reduction of vimentin, TCF/Zeb1, and N-cadherin protein expression was detected in the TPC-1 cells.Conclusions: Increased activation of AMPK in PTC cell lines leads to a strong antitumor response, as measured by the inhibition of cell proliferation, cell migration, and induction of cell death. AMPK activation also reverses EMT in TPC-1 cells.