GANT61 and Lithium Chloride Inhibit the Growth of Head and Neck Cancer Cell Lines Through the Regulation of GLI3 Processing by GSK3β
Abstract
:1. Introduction
2. Results
2.1. The HH-GLI Signaling Pathway Is Active in HNSCC Cell Lines
2.2. HNSCC Cell Lines Respond to Downstream Inhibition More Efficiently Than to Upstream Inhibition
2.3. GANT61 and LiCl Regulate GLI3 Protein Levels in HNSCC
2.4. The Effect of LiCl Inhibition is Mediated by GLI3 Processing by GSK3β
2.5. Downstream Inhibition Affects Colony Forming and Wound Closure Capabilities of HNSCC Cell Lines
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Cell Assays
4.2. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)
4.3. Western Blotting
4.4. Immunoprecipitation and Coomassie Staining
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
HH-GLI | Hedgehog-Gli |
LiCl | Lithium chloride |
HNSCC | Head and neck squamous cell carcinoma |
HPV16 | Human papilloma virus 16 |
CSC | Cancer stem cells |
EGFR | Epidermal growth factor receptor |
PI3K | Phosphoinositol 3 kinase |
VEGF | Vascular endothelial growth factor |
PTCH1 | Patched 1 |
HH | Hedgehog |
SMO | Smoothened |
βTrCP1 | Beta-transducin repeat containing protein 1 |
GSK3β | Glycogen synthase 3 beta |
PKA | Protein kinase A |
SUFU | Suppressor of fused |
CK1 | Casein kinase 1 |
GLI | Glioma-associated oncogene homolog |
EMT | Epithelial-mesenchymal transition |
NQC | Nanoquinacrine |
DMEM:HAM’S | Dulbecco’s Modified Eagle Medium: Ham’s F12 |
EMEM | Eagle’s Minimum Essential Medium |
FBS | Fetal bovine Serum |
MTT | 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide |
PBS | Phosphate Buffered Saline |
RIPA | Radioimmunoprecipitation assay buffer |
BCA | Bicinchonic Acid |
PAA | Polyacrylamide |
TBST | Tris-Buffered Saline, 0.1% Tween® 20 Detergent |
HRP | Horseradish Peroxidase |
TENN | 50 mM Tris, 5 mM EDTA, 150 mM NaCl, 0.5% NP-40, pH 8.0 |
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Line | Tissue | Cyclopamine (µM) | LiCl (mM) | GANT61 (μM) |
---|---|---|---|---|
SCC9 | Tongue | >10 | 11.45 | >25 |
SCC25 | Tongue | 9.51 | 15.24 | 6.83 |
A253 | Salivary gland | 6.48 | 14.56 | 8.36 |
DETROIT562 | Pharynx | >10 | 11.4 | 9.54 |
FADU | Hypopharynx | >10 | 5.53 | 19.7 |
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Zubčić, V.; Rinčić, N.; Kurtović, M.; Trnski, D.; Musani, V.; Ozretić, P.; Levanat, S.; Leović, D.; Sabol, M. GANT61 and Lithium Chloride Inhibit the Growth of Head and Neck Cancer Cell Lines Through the Regulation of GLI3 Processing by GSK3β. Int. J. Mol. Sci. 2020, 21, 6410. https://doi.org/10.3390/ijms21176410
Zubčić V, Rinčić N, Kurtović M, Trnski D, Musani V, Ozretić P, Levanat S, Leović D, Sabol M. GANT61 and Lithium Chloride Inhibit the Growth of Head and Neck Cancer Cell Lines Through the Regulation of GLI3 Processing by GSK3β. International Journal of Molecular Sciences. 2020; 21(17):6410. https://doi.org/10.3390/ijms21176410
Chicago/Turabian StyleZubčić, Vedran, Nikolina Rinčić, Matea Kurtović, Diana Trnski, Vesna Musani, Petar Ozretić, Sonja Levanat, Dinko Leović, and Maja Sabol. 2020. "GANT61 and Lithium Chloride Inhibit the Growth of Head and Neck Cancer Cell Lines Through the Regulation of GLI3 Processing by GSK3β" International Journal of Molecular Sciences 21, no. 17: 6410. https://doi.org/10.3390/ijms21176410