Deguelin Potentiates Apoptotic Activity of an EGFR Tyrosine Kinase Inhibitor (AG1478) in PIK3CA-Mutated Head and Neck Squamous Cell Carcinoma
Abstract
:1. Introduction
2. Results
2.1. Head and Neck Squamous Cell Carcinoma (HNSCC) Cell Lines after AG1478 Treatment
2.1.1. AG1478 Suppressed the Phosphorylation of EGFR in a Dose-Dependent Manner in HSC4 Cells
2.1.2. AG1478 Did Not Induce Apoptosis in HSC-4 (Mutant PIK3CA), but Induced Apoptosis in Ca9-22 Cells (WT PIK3CA)
2.1.3. While AG1478 Suppressed the Expression of Both Phosphorylated ERK and AKT in Ca9-22 Cells (WT PIK3CA), AG1478 Suppressed the Expression of Phosphorylated ERK and Did Not Significantly Suppress the Expression of Phosphorylated AKT in HSC-4 Cells (Mutant PIK3CA)
2.1.4. The Sensitivity of HNSCC Cells to AG1478 Depended on PIK3CA Gene Status
2.2. Deguelin Suppressed Phosphorylation Level of AKT in Dose- and Time-Dependent Manner in HSC-4 Cells
2.3. The Combination of AG1478 and Deguelin in HSC-4 Cells
2.3.1. Combination of AG1478 and Deguelin Was Stronger Than AG1478 Alone with Regard to the Effect on the Active Form of AKT, and the Combination Was Stronger than Deguelin Alone with Regard to the Effect on the Active Form of ERK in HSC-4 Cells
2.3.2. The Combination of Deguelin and AG1478 Significantly Induced Apoptosis Relative to the Control or Alone in HSC-4 Cells
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell and Cell Cultures
4.3. Cell Viability Assay
4.4. Protein Preparation and Western Blot Analysis
4.5. Vectors
4.6. Vector Construction and Transfection
4.7. Protein Assay
4.8. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Saranath, D.; Panchal, R.G.; Nair, R.; Mehta, A.R.; Sanghavi, V.D.; Deo, M.G. Amplification and overexpression of epidermal growth factor receptor gene in human oropharyngeal cancer. Eur. J. Cancer B Oral. Oncol. 1992, 28, 139–143. [Google Scholar] [CrossRef]
- Burtness, B. The role of cetuximab in the treatment of squamous cell cancer of the head and neck. Expert Opin. Biol. Ther. 2005, 5, 1085–1093. [Google Scholar] [CrossRef] [PubMed]
- Yun, C.H.; Boggon, T.J.; Li, Y.; Woo, M.S.; Greulich, H.; Meyerson, M.; Eck, M.J. Structures of lung cancer-derived EGFR mutants and inhibitor complexes: Mechanism of activation and insights into differential inhibitor sensitivity. Cancer Cell 2007, 11, 217–227. [Google Scholar] [CrossRef] [PubMed]
- Mitsudomi, T.; Yatabe, Y. Mutations of the epidermal growth factor receptor gene and related genes as determinants of epidermal growth factor receptor tyrosine kinase inhibitors sensitivity in lung cancer. Cancer Sci. 2007, 98, 1817–1824. [Google Scholar] [CrossRef] [PubMed]
- Sordella, R.; Bell, D.W.; Haber, D.A.; Settleman, J. Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science 2004, 305, 1163–1167. [Google Scholar] [CrossRef] [PubMed]
- Yun, C.H.; Mengwasser, K.E.; Toms, A.V.; Woo, M.S.; Greulich, H.; Wong, K.K.; Meyerson, M.; Eck, M.J. The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP. Proc. Natl. Acad. Sci. USA 2008, 105, 2070–2075. [Google Scholar] [CrossRef] [PubMed]
- Baba, Y.; Fujii, M.; Tokumaru, Y.; Kato, Y. Present and Future of EGFR Inhibitors for Head and Neck Squamous Cell Cancer. J. Oncol. 2012, 2012, 986725. [Google Scholar] [CrossRef] [PubMed]
- Cohen, E.E.; Rosen, F.; Stadler, W.M.; Recant, W.; Stenson, K.; Huo, D.; Vokes, E.E. Phase II trial of ZD1839 in recurrent or metastatic squamous cell carcinoma of the head and neck. J. Clin. Oncol. 2003, 21, 1980–1987. [Google Scholar] [CrossRef] [PubMed]
- Soulieres, D.; Senzer, N.N.; Vokes, E.E.; Hidalgo, M.; Agarwala, S.S.; Siu, L.L. Multicenter phase II study of erlotinib, an oral epidermal growth factor receptor tyrosine kinase inhibitor, in patients with recurrent or metastatic squamous cell cancer of the head and neck. J. Clin. Oncol. 2004, 22, 77–85. [Google Scholar] [CrossRef] [PubMed]
- Khattri, A.; Zuo, Z.; Bragelmann, J.; Keck, M.K.; El Dinali, M.; Brown, C.D.; Stricker, T.; Munagala, A.; Cohen, E.E.; Lingen, M.W.; et al. Rare occurrence of EGFRvIII deletion in head and neck squamous cell carcinoma. Oral Oncol. 2015, 51, 53–58. [Google Scholar] [CrossRef] [PubMed]
- Eberhard, D.A.; Johnson, B.E.; Amler, L.C.; Goddard, A.D.; Heldens, S.L.; Herbst, R.S.; Ince, W.L.; Jänne, P.A.; Januario, T.; Johnson, D.H.; et al. Mutations in the epidermal growth factor receptor and in KRAS are predictive and prognostic indicators in patients with non-small-cell lung cancer treated with chemotherapy alone and in combination with erlotinib. J. Clin. Oncol. 2005, 23, 5900–5909. [Google Scholar] [CrossRef] [PubMed]
- Anderson, J.A.; Irish, J.C.; McLachlin, C.M.; Ngan, B.Y. H-ras oncogene mutation and human papillomavirus infection in oral carcinomas. Arch. Otolaryngol. Head Neck Surg. 1994, 120, 755–760. [Google Scholar] [CrossRef] [PubMed]
- Qiu, W.; Schönleben, F.; Li, X.; Ho, D.J.; Close, L.G.; Manolidis, S.; Bennett, B.P.; Su, G.H. PIK3CA mutations in head and neck squamous cell carcinoma. Clin. Cancer Res. 2006, 12, 1441–1446. [Google Scholar] [CrossRef] [PubMed]
- Kozaki, K.; Imoto, I.; Pimkhaokham, A.; Hasegawa, S.; Tsuda, H.; Omura, K.; Inazawa, J. PIK3CA mutation is an oncogenic aberration at advanced stages of oral squamous cell carcinoma. Cancer Sci. 2006, 97, 1351–1358. [Google Scholar] [CrossRef] [PubMed]
- Mriouah, J.; Boura, C.; Pinel, S.; Chretien, A.S.; Fifre, A.; Merlin, J.L.; Faivre, B. Cellular response to cetuximab in PTEN-silenced head and neck squamous cell carcinoma cell line. Int. J. Oncol. 2010, 37, 1555–1563. [Google Scholar] [PubMed]
- Murillo, G.; Kosmeder, J.W., 2nd; Pezzuto, J.M.; Mehta, R.G. Deguelin suppresses the formation of carcinogen-induced aberrant crypt foci in the colon of CF-1 mice. Int. J. Cancer 2003, 104, 7–11. [Google Scholar] [CrossRef] [PubMed]
- Gerhäuser, C.; Mar, W.; Lee, S.K.; Suh, N.; Luo, Y.; Kosmeder, J.; Luyengi, L.; Fong, H.H.; Kinghorn, A.D.; Moriarty, R.M.; et al. Rotenoids mediate potent cancer chemopreventive activity through transcriptional regulation of ornithine decarboxylase. Nat. Med. 1995, 1, 260–266. [Google Scholar] [CrossRef] [PubMed]
- Udeani, G.O.; Gerhauser, C.; Thomas, C.F.; Moon, R.C.; Kosmeder, J.W.; Kinghorn, A.D.; Moriarty, R.M.; Pezzuto, J.M. Cancer chemopreventive activity mediated by deguelin, a naturally occurring rotenoid. Cancer Res. 1997, 57, 3424–3428. [Google Scholar] [PubMed]
- Lee, H.Y.; Oh, S.H.; Woo, J.K.; Kim, W.Y.; van Pelt, C.S.; Price, R.E.; Cody, D.; Tran, H.; Pezzuto, J.M.; Moriarty, R.M.; et al. Chemopreventive effects of deguelin, a novel Akt inhibitor, on tobacco-induced lung tumorigenesis. J. Natl. Cancer Inst. 2005, 97, 1695–1699. [Google Scholar] [CrossRef] [PubMed]
- Yang, Y.L.; Ji, C.; Bi, Z.G.; Lu, C.C.; Wang, R.; Gu, B.; Cheng, L. Deguelin induces both apoptosis and autophagy in cultured head and neck squamous cell carcinoma cells. PLoS ONE 2013, 8, e54736. [Google Scholar] [CrossRef] [PubMed]
- Baba, Y.; Fujii, M.; Maeda, T.; Suzuki, A.; Yuzawa, S.; Kato, Y. Deguelin Induces Apoptosis by Targeting Both EGFR-Akt and IGF1R-Akt Pathways in Head and Neck Squamous Cell Cancer Cell Lines. BioMed Res. Int. 2015, 2015, 657179. [Google Scholar] [CrossRef] [PubMed]
- Sheikh Ali, M.A.; Gunduz, M.; Nagatsuka, H.; Gunduz, E.; Cengiz, B.; Fukushima, K.; Beder, L.B.; Demircan, K.; Fujii, M.; Yamanaka, N.; et al. Expression and mutation analysis of epidermal growth factor receptor in head and neck squamous cell carcinoma. Cancer Sci. 2008, 99, 1589–1594. [Google Scholar] [PubMed]
- Liu, Y.P.; Lee, J.J.; Lai, T.C.; Lee, C.H.; Hsiao, Y.W.; Chen, P.S.; Liu, W.T.; Hong, C.Y.; Lin, S.K.; Ping Kuo, M.Y.; et al. Suppressive function of low-dose deguelin on the invasion of oral cancer cells by downregulating tumor necrosis factor α-induced nuclear factor-kappa B signaling. Head Neck 2016, 38, E524–E534. [Google Scholar] [CrossRef] [PubMed]
- Rebucci, M.; Peixoto, P.; Dewitte, A.; Wattez, N.; De Nuncques, M.A.; Rezvoy, N.; Vautravers-Dewas, C.; Buisine, M.P.; Guerin, E.; Peyrat, J.P.; et al. Mechanisms underlying resistance to cetuximab in the HNSCC cell line: Role of AKT inhibition in bypassing this resistance. Int. J. Oncol. 2011, 38, 189–200. [Google Scholar] [PubMed]
- D’Amato, V.; Rosa, R.; D’Amato, C.; Formisano, L.; Marciano, R.; Nappi, L.; Raimondo, L.; di Mauro, C.; Servetto, A.; Fusciello, C.; et al. The dual PI3K/mTOR inhibitor PKI-587 enhances sensitivity to cetuximab in EGFR-resistant human head and neck cancer models. Br. J. Cancer 2014, 110, 2887–2895. [Google Scholar] [CrossRef] [PubMed]
- Massarelli, E.; Lin, H.; Ginsberg, L.E.; Tran, H.T.; Lee, J.J.; Canales, J.R.; Williams, M.D.; Blumenschein, G.R., Jr.; Lu, C.; Heymach, J.V.; et al. Phase II trial of everolimus and erlotinib in patients with platinum-resistant recurrent and/or metastatic head and neck squamous cell carcinoma. Ann. Oncol. 2015, 26, 1476–1480. [Google Scholar] [CrossRef] [PubMed]
- Bauman, J.E.; Arias-Pulido, H.; Lee, S.J.; Fekrazad, M.H.; Ozawa, H.; Fertig, E.; Howard, J.; Bishop, J.; Wang, H.; Olson, G.T.; et al. A phase II study of temsirolimus and erlotinib in patients with recurrent and/or metastatic, platinum-refractory head and neck squamous cell carcinoma. Oral Oncol. 2013, 49, 461–467. [Google Scholar] [CrossRef] [PubMed]
- Loeffler-Ragg, J.; Witsch-Baumgartner, M.; Tzankov, A.; Hilbe, W.; Schwentner, I.; Sprinzl, G.M.; Utermann, G.; Zwierzina, H. Low incidence of mutations in EGFR kinase domain in Caucasian patients with head and neck squamous cell carcinoma. Eur. J. Cancer 2006, 42, 109–111. [Google Scholar] [CrossRef] [PubMed]
- Bouali, S.; Chretien, A.S.; Ramacci, C.; Rouyer, M.; Becuwe, P.; Merlin, J.L. PTEN expression controls cellular response to cetuximab by mediating PI3K/AKT and RAS/RAF/MAPK downstream signaling in KRAS wild-type, hormone refractory prostate cancer cells. Oncol. Rep. 2009, 21, 731–735. [Google Scholar] [PubMed]
- Hirose, K.; Isogai, E.; Mizugai, H.; Ueda, I. Adhesion of Porphyromonas gingivalis fimbriae to human gingival cell line Ca9-22. Oral Microbiol. Immunol. 1996, 11, 402–406. [Google Scholar] [CrossRef] [PubMed]
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Baba, Y.; Maeda, T.; Suzuki, A.; Takada, S.; Fujii, M.; Kato, Y. Deguelin Potentiates Apoptotic Activity of an EGFR Tyrosine Kinase Inhibitor (AG1478) in PIK3CA-Mutated Head and Neck Squamous Cell Carcinoma. Int. J. Mol. Sci. 2017, 18, 262. https://doi.org/10.3390/ijms18020262
Baba Y, Maeda T, Suzuki A, Takada S, Fujii M, Kato Y. Deguelin Potentiates Apoptotic Activity of an EGFR Tyrosine Kinase Inhibitor (AG1478) in PIK3CA-Mutated Head and Neck Squamous Cell Carcinoma. International Journal of Molecular Sciences. 2017; 18(2):262. https://doi.org/10.3390/ijms18020262
Chicago/Turabian StyleBaba, Yuh, Toyonobu Maeda, Atsuko Suzuki, Satoshi Takada, Masato Fujii, and Yasumasa Kato. 2017. "Deguelin Potentiates Apoptotic Activity of an EGFR Tyrosine Kinase Inhibitor (AG1478) in PIK3CA-Mutated Head and Neck Squamous Cell Carcinoma" International Journal of Molecular Sciences 18, no. 2: 262. https://doi.org/10.3390/ijms18020262