新型热休克蛋白90（HSP90）抑制剂可有效靶向甲状腺癌干细胞的功能，阻止其迁移和侵袭。
Novel HSP90 inhibitors effectively target functions of thyroid cancer stem cell preventing migration and invasion.
作者信息
White Peter T, Subramanian Chitra, Zhu Qing, Zhang Huaping, Zhao Huiping, Gallagher Robert, Timmermann Barbara N, Blagg Brian S J, Cohen Mark S
机构信息
Department of Surgery, University of Michigan, Ann Arbor, MI.
Department of Medicinal Chemistry, The University of Kansas, Lawrence, KS.
出版信息
Surgery. 2016 Jan;159(1):142-51. doi: 10.1016/j.surg.2015.07.050. Epub 2015 Nov 2.
BACKGROUND
Thyroid cancer stem cells (CSCs) with ALDH and CD44 markers contribute to tumor growth and aggressiveness. We hypothesized that novel HSP90 inhibitors (KU711, WGA-TA) and 17-AAG can effectively target the function of thyroid CSCs in vitro and prevent migration and invasion.
METHODS
Validated papillary (TPC1), follicular (FTC238,WRO), and anaplastic (ACT1) human thyroid cancer cell lines were treated with 3 HSP90 inhibitors. CSCs were quantified for aldehyde dehydrogenase by flow cytometry, CD44 expression by Western blot, and thyrosphere formation assay. Cellular pathway proteins were analyzed by Western blot and migration/invasion by Boyden-chambers.
RESULTS
WGA-TA and 17-AAG induced HSP70 compensation (not observed with KU711) on Western blot in all cell lines (>1,000 fold vs controls). Only WGA-TA degraded HSP90-Cdc37 complexing by 60-70% versus controls. Expression of HSP90 clients β-catenin, BRAF, Akt, and phospho-Akt were significantly inhibited by WGA-TA treatment (50-80%, 50-90%, >80%, and >90%) compared with controls, KU711, and 17-AAG treatment. KU711 and WGA-TA decreased CD44 expression in all cell lines (25-60% vs controls/17-AAG), decreased ALDEFLOR activity by 69-98% (P < .005), and decreased sphere formation by 64-99% (P < .05 each). Finally, cell migration was decreased by 31-98%, 100%, and 30-38%, and invasion by 75-100%, 100%, and 47% by KU711,WGA-TA, and 17-AAG treatment (P < .05) each, respectively.
CONCLUSION
KU711 and WGA-TA are novel HSP90 inhibitors targeting CSC function and inhibiting cell migration/invasion in differentiated and anaplastic thyroid cancers, warranting further translational evaluation in vivo.
背景
具有乙醛脱氢酶（ALDH）和CD44标志物的甲状腺癌干细胞（CSCs）促进肿瘤生长和侵袭性。我们假设新型热休克蛋白90（HSP90）抑制剂（KU711、WGA-TA）和17-烯丙基氨基-17-去甲氧基格尔德霉素（17-AAG）可在体外有效靶向甲状腺癌干细胞的功能并阻止其迁移和侵袭。
方法
用3种HSP90抑制剂处理经过验证的人乳头状（TPC1）、滤泡状（FTC238、WRO）和间变性（ACT1）甲状腺癌细胞系。通过流式细胞术对醛脱氢酶进行定量分析，通过蛋白质免疫印迹法检测CD44表达，并进行甲状腺球形成试验。通过蛋白质免疫印迹法分析细胞信号通路蛋白，通过博伊登小室检测细胞迁移/侵袭能力。
结果
蛋白质免疫印迹法显示，WGA-TA和17-AAG在所有细胞系中均诱导了热休克蛋白70（HSP70）代偿（KU711未观察到）（与对照相比增加超过1000倍）。与对照相比，只有WGA-TA使HSP90-细胞分裂周期蛋白37（Cdc37）复合物降解60%-70%。与对照、KU711和17-AAG处理相比，WGA-TA处理显著抑制了HSP90客户蛋白β-连环蛋白、BRAF、蛋白激酶B（Akt）和磷酸化Akt的表达（分别为50%-80%、50%-90%、>80%和>90%）。KU711和WGA-TA降低了所有细胞系中CD44的表达（与对照/17-AAG相比降低25%-60%），使醛荧光素活性降低69%-98%（P <.005），使球体形成减少64%-99%（各P <.05）。最后，KU711、WGA-TA和17-AAG处理分别使细胞迁移减少31%-98%、100%和30%-38%，使侵袭减少75%-100%、100%和47%（各P <.05）。
结论
KU711和WGA-TA是新型HSP90抑制剂，可靶向癌干细胞功能并抑制分化型和间变性甲状腺癌的细胞迁移/侵袭，值得在体内进行进一步的转化评估。
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