The AKT antagonist AZD5363 suppresses features associated with cancer progression in human larynx cancer cells

Abstract

Objectives: Larynx cancer (LCa) represents approximately 30% of all cancers seen in the head and neck region, with an unchanged overall survival rate over the last decades. Although several novel diagnostic and therapeutic options has been developed, an effective treatment strategy is not currently available due to the high metastatic and recurrent potential of LCa. In this study, we aimed at investigating the inhibitory potential of AZD5363 on the phenotypes associated with LCa progression in vitro.

Methods: The impacts of AZD5363 on the proliferation, colony formation, and apoptosis potentials of HEp-2 cells were tested using Cell Viability Detection Kit-8, soft agar assay and Annexin V-FITC Apoptosis assay, respectively. Migration features of cells were evaluated using scratch and transwell migration assays.

Results: We showed that AZD5363 increased phosphorylation of AKT and inhibited the phosphorylation of its downstream effector GSK3β in an in vitro LCa model in line with the findings of previous studies carried out with different cancer types. Besides, AZD5363 successfully suppressed proliferative, clonogenic, and migratory features of HEp-2 cells through induction of apoptosis.

Conclusions: We revealed putative functions of AZD5363 in vitro that points its potential to be used as an adjuvant agent against LCa. However, further comprehensive molecular and clinical research is needed to elucidate the potential use of AZD5363 in LCa therapy in detail. 

Keywords

• Larynx cancer
• AZD5363
• AKT
• chemotherapy

References

1. 1. Armstrong WB, Vokes DE, Verma SP. Malignant tumors of the larynx. In: Flint PW, Haughey BH, Lund V, Niparko JK, Robbins KT, Tomas JP, et al., eds. Cummings Otolaryngology - Head and Neck Surgery. 6th ed., Philadelphia, PA: Elsevier Inc., Sander; 2015: pp.1601-33.e10.
2. 2. Yilmaz SS, Guzel E, Karatas OF, Yilmaz M, Creighton CJ, Ozen M. MiR-221 as a pre- and postoperative plasma biomarker for larynx cancer patients. Laryngoscope 2015;125:E377-81.
3. 3. Jaipuria B, Dosemane D, Kamath PM, Sreedharan SS, Shenoy VS. Staging of Laryngeal and Hypopharyngeal Cancer: Computed Tomography versus Histopathology. Iran J Otorhinolaryngol 2018;30:189-94.
4. 4. Zhang Y, Hu H. Long non-coding RNA CCAT1/miR-218/ZFX axis modulates the progression of laryngeal squamous cell cancer. Tumour Biol 2017;39:1010428317699417.
5. 5. Yu Q, Zhang X, Ji C, Yang H, Gao M, Hong S, et al. Survival analysis of laryngeal carcinoma without laryngectomy, radiotherapy, or chemotherapy. Eur Arch Otorhinolaryngol 2012;269:2103-9.
6. 6. Wu Y, Zhang Y, Niu M, Shi Y, Liu H, Yang D, et al. Whole-transcriptome analysis of CD133+CD144+ cancer stem cells derived from human laryngeal squamous cell carcinoma cells. Cell Physiol Biochem 2018;47:1696-710.
7. 7. Cantley LC. The phosphoinositide 3-kinase pathway. Science 2002;296:1655-7.
8. 8. Gupta AK, McKenna WG, Weber CN, Feldman MD, Goldsmith JD, Mick R, et al. Local recurrence in head and neck cancer: relationship to radiation resistance and signal transduction. Clin Cancer Res 2002;8:885-92.

9. 9. Pérez-Tenorio G, Alkhori L, Olsson B, Waltersson MA, Nordenskjöld B, Rutqvist LE, et al. PIK3CA mutations and PTEN loss correlate with similar prognostic factors and are not mutually exclusive in breast cancer. Clin Cancer Res 2007;13:3577-84.
10. 10. Harima Y, Sawada S, Nagata K, Sougawa M, Ostapenko V, Ohnishi T. Mutation of the PTEN gene in advanced cervical cancer correlated with tumor progression and poor outcome after radiotherapy. Int J Oncol 2001;18:493-7.
11. 11. Bedolla R, Prihoda TJ, Kreisberg JI, Malik SN, Krishnegowda NK, Troyer DA, et al. Determining risk of biochemical recurrence in prostate cancer by immunohistochemical detection of PTEN expression and Akt activation. Clin Cancer Res 2007;13:3860-7.
12. 12. Snietura M, Jaworska M, Mlynarczyk-Liszka J, Goraj-Zajac A, Piglowski W, Lange D, et al. PTEN as a prognostic and predictive marker in postoperative radiotherapy for squamous cell cancer of the head and neck. PLoS One 2012;7:e33396.
13. 13. Liu P, Cheng H, Roberts TM, Zhao JJ. Targeting the phosphoinositide 3-kinase pathway in cancer. Nat Rev Drug Discov 2009;8:627-44.
14. 14. Bellacosa A, Kumar CC, Di Cristofano A, Testa JR. Activation of AKT kinases in cancer: implications for therapeutic targeting. Adv Cancer Res 2005;94:29-86.
15. 15. Altomare DA, Testa JR. Perturbations of the AKT signaling pathway in human cancer. Oncogene 2005;24:7455-64.
16. 16. Chen Y, He Y, Zhang S, Li L, Zhu X, Liu Y. [The expression of oncogene AKT2 in laryngeal squamous cell carcinoma and its clinical significance]. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2009;23:539-541, 545. [Article in Chinese]
17. 17. Dionysopoulos D, Pavlakis K, Kotoula V, Fountzilas E, Markou K, Karasmanis U, et al. Cyclin D1, EGFR, and Akt/mTOR pathway. Potential prognostic markers in localized laryngeal squamous cell carcinoma. Strahlenther Onkol 2013;189:202-14.
18. 18. Crabb SJ, Birtle AJ, Martin K, Downs N, Ratcliffe I, Maishman T, et al. ProCAID: a phase I clinical trial to combine the AKT inhibitor AZD5363 with docetaxel and prednisolone chemotherapy for metastatic castration resistant prostate cancer. Invest New Drugs 2017;35:599-607.
19. 19. Davies BR, Greenwood H, Dudley P, Crafter C, Yu D-H, Zhang J, et al. Preclinical pharmacology of AZD5363, an inhibitor of AKT: pharmacodynamics, antitumor activity, and correlation of monotherapy activity with genetic background. Mol Cancer Ther 2012;11:873-87.
20. 20. Toren P, Kim S, Cordonnier T, Crafter C, Davies BR, Fazli L, et al. Combination AZD5363 with enzalutamide significantly delays enzalutamide-resistant prostate cancer in preclinical models. Eur Urol 2015;67:986-90.
21. 21. Lamoureux F, Thomas C, Crafter C, Kumano M, Zhang F, Davies BR, et al. Blocked autophagy using lysosomotropic agents sensitizes resistant prostate tumor cells to the novel Akt inhibitor AZD5363. Clin Cancer Res 2013;19:833-44.
22. 22. Feng S, Shao L, Castro P, Coleman I, Nelson PS, Smith PD, et al. Combination treatment of prostate cancer with FGF receptor and AKT kinase inhibitors. Oncotarget 2017;8:6179-92.
23. 23. De Velasco MA, Kura Y, Yoshikawa K, Nishio K, Davies BR, Uemura H. Efficacy of targeted AKT inhibition in genetically engineered mouse models of PTEN-deficient prostate cancer. Oncotarget 2016;7:15959-76.
24. 24. Wang Y, Zhao Y, Liu Y, Tian L, Jin D. Chamaejasmine inactivates Akt to trigger apoptosis in human HEp-2 larynx carcinoma cells. Molecules 2011;16:8152-64.
25. 25. Steuer CE, El-Deiry M, Parks JR, Higgins KA, Saba NF. An update on larynx cancer. CA Cancer J Clin. 2017;67:31-50.
26. 26. Adelstein DJ, Li Y, Adams GL, Wagner H Jr, Kish JA, Ensley JF, et al. An intergroup phase III comparison of standard radiation therapy and two schedules of concurrent chemoradiotherapy in patients with unresectable squamous cell head and neck cancer. J Clin Oncol 2003;21:92-8.
27. 27. Haigentz M Jr, Silver CE, Hartl DM, Takes RP, Rodrigo JP, Robbins KT, et al. Chemotherapy regimens and treatment protocols for laryngeal cancer. Expert Opin Pharmacother 2010;11:1305-16.
28. 28. Polivka J, Janku F. Molecular targets for cancer therapy in the PI3K/AKT/mTOR pathway. Pharmacol Ther 2014;142:164-75.
29. 29. Zhang Y, Zheng Y, Faheem A, Sun T, Li C, Li Z, et al. A novel AKT inhibitor, AZD5363, inhibits phosphorylation of AKT downstream molecules, and activates phosphorylation of mTOR and SMG-1 dependent on the liver cancer cell type. Oncol Lett 2016;11:1685-92.
30. 30. Ribas R, Pancholi S, Guest SK, Marangoni E, Gao Q, Thuleau A, et al. AKT Antagonist AZD5363 influences estrogen receptor function in endocrine-resistant breast cancer and synergizes with fulvestrant (ICI182780) in vivo. Mol Cancer Ther 2015;14:2035-48.
31. 31. Tamura K, Hashimoto J, Tanabe Y, Kodaira M, Yonemori K, Seto T, et al. Safety and tolerability of AZD5363 in Japanese patients with advanced solid tumors. Cancer Chemother Pharmacol.2016;77:787-95.
32. 32. Choi AR, Kim JH, Woo YH, Cheon JH, Kim HS, Yoon S. Co-treatment of LY294002 or MK-2206 with AZD5363 attenuates AZD5363-induced increase in the level of phosphorylated AKT. Anticancer Res 2016;36:5849-58.