TY - JOUR
T1 - Novel HSP90 inhibitors effectively target functions of thyroid cancer stem cell preventing migration and invasion
AU - White, Peter T.
AU - Subramanian, Chitra
AU - Zhu, Qing
AU - Zhang, Huaping
AU - Zhao, Huiping
AU - Gallagher, Robert
AU - Timmermann, Barbara N.
AU - Blagg, Brian S.J.
AU - Cohen, Mark S.
N1 - Funding Information:
This work was partly funded by the National Institutes of Health ( T32 CA009672 , R01 CA173292 , R01 CA120458 ), IND 0061464 from the Kansas Biosciences Authority (KBA) and Center for Heartland Plant Innovations (HPI), RIC 2506007-700 from the KU Research Innovation Fund, NSF MRI Grant #0320648 , NIH Shared Instrumentation Grant # S10RR024664 and S10RR014767 , the University of Michigan Comprehensive Cancer Center Support Grant ( P30 CA046592 ), and the University of Michigan Department of Surgery.
Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2016/1/1
Y1 - 2016/1/1
N2 - 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.
AB - 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.
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U2 - 10.1016/j.surg.2015.07.050
DO - 10.1016/j.surg.2015.07.050
M3 - Article
C2 - 26542767
AN - SCOPUS:84951779487
SN - 0039-6060
VL - 159
SP - 142
EP - 151
JO - Surgery (United States)
JF - Surgery (United States)
IS - 1
ER -