Analysis of the Epidermal Growth Factor Receptor/phosphoinositide-dependent Protein Kinase-1 Axis in Tumor of the External Auditory Canal in Response to Epidermal Growth Factor Stimulation

Overview

Journal Laryngoscope Investig Otolaryngol

Publisher Wiley

Specialty Otorhinolaryngology

Date 2022 Jun 23

PMID 35734041

Authors

Naotaro Akiyama

Tomomi Yamamoto-Fukuda

Mamoru Yoshikawa

Hiromi Kojima

Affiliations

Soon will be listed here.

Abstract

Objectives: The epidermal growth factor receptor (EGFR) is related to the invasion and metastasis of external auditory canal (EAC) squamous cell carcinoma (SCC). The phosphoinositide-dependent protein kinase-1 (PDPK1) accelerates tumor cell growth through anti-apoptotic signaling under the influence of downstream EGFR-mediated signaling pathways. In this study, we investigated the EGFR/PDPK1 axis in the EAC under EGF stimulation.

Methods: We confirmed EGFR and PDPK1 expression in human EACSCC specimens immunohistochemically. We next transfected the EGF expression vector in the mouse EAC and then conducted a PDPK1 inhibitory experiment. Immunohistochemical analysis was performed in the mouse EAC, using anti-EGF, anti-EGFR, anti-PDPK1, and anti-Ki67 antibodies. Immunohistochemical analysis of cleaved caspase-3 and terminal deoxy(d)-UTP nick end labeling (TUNEL) detection assays were also performed for the assessment of apoptosis in the inhibitory experiment.

Results: Immunohistochemical analysis revealed overexpression and colocalization of EGFR and PDPK1 in human EACSCC specimens. The growth of a protuberant tumor was observed in the mouse EAC in which EGF expression vector was transfected, and EGF, EGFR, PDPK1, and Ki67 labeling indexes (LIs) were significantly increased. PDPK1 inhibition then induced normal epithelial appearance in the EAC. Moreover, EGF, EGFR, PDPK1, and Ki67 LIs were decreased, and cleaved caspase-3 and TUNEL LIs were increased in the EAC.

Conclusion: We demonstrated the possibility that PDPK1 plays an important role in EACSCC.Level of Evidence: NA.

Citing Articles

Analysis of L1 Cell Adhesion Molecule and Fucosyltransferase 8 Expression in Cells After Stretch and Human EACSCC Tissue.

Akiyama N, Yamamoto-Fukuda T, Kojima H J Int Adv Otol. 2025; 21(1):1-8.

PMID: 39936830 PMC: 11843287. DOI: 10.5152/iao.2025.241652.

Analysis of the epidermal growth factor receptor/phosphoinositide-dependent protein kinase-1 axis in tumor of the external auditory canal in response to epidermal growth factor stimulation.

Akiyama N, Yamamoto-Fukuda T, Yoshikawa M, Kojima H Laryngoscope Investig Otolaryngol. 2022; 7(3):730-739.

PMID: 35734041 PMC: 9195017. DOI: 10.1002/lio2.785.

References

Zhou W, Wu G, Huang J, Li J, Hao C, He Q . Low expression of PDK1 inhibits renal cell carcinoma cell proliferation, migration, invasion and epithelial mesenchymal transition through inhibition of the PI3K-PDK1-Akt pathway. Cell Signal. 2018; 56:1-14. DOI: 10.1016/j.cellsig.2018.11.016. View

Sato K, Komune N, Hongo T, Koike K, Niida A, Uchi R . Genetic landscape of external auditory canal squamous cell carcinoma. Cancer Sci. 2020; 111(8):3010-3019. PMC: 7419060. DOI: 10.1111/cas.14515. View

Ruicci K, Plantinga P, Pinto N, Khan M, Stecho W, Dhaliwal S . Disruption of the RICTOR/mTORC2 complex enhances the response of head and neck squamous cell carcinoma cells to PI3K inhibition. Mol Oncol. 2019; 13(10):2160-2177. PMC: 6763779. DOI: 10.1002/1878-0261.12558. View

Akiyama N, Yamamoto-Fukuda T, Yoshikawa M, Kojima H . Regulation of DNA methylation levels in the process of oral mucosal regeneration in a rat oral ulcer model. Histol Histopathol. 2019; 35(3):247-256. DOI: 10.14670/HH-18-147. View

Gagliardi P, di Blasio L, Orso F, Seano G, Sessa R, Taverna D . 3-phosphoinositide-dependent kinase 1 controls breast tumor growth in a kinase-dependent but Akt-independent manner. Neoplasia. 2012; 14(8):719-31. PMC: 3431179. DOI: 10.1593/neo.12856. View

Ericson K, Gan C, Cheong I, Rago C, Samuels Y, Velculescu V . Genetic inactivation of AKT1, AKT2, and PDPK1 in human colorectal cancer cells clarifies their roles in tumor growth regulation. Proc Natl Acad Sci U S A. 2010; 107(6):2598-603. PMC: 2823889. DOI: 10.1073/pnas.0914018107. View

Maki D, Okami K, Ebisumoto K, Sakai A, Hamada M, Ogura G . Immunohistochemical Marker Expression in Temporal Bone Squamous Cell Carcinoma. Tokai J Exp Clin Med. 2021; 46(2):89-93. View

Pollock N, Grandis J . HER2 as a therapeutic target in head and neck squamous cell carcinoma. Clin Cancer Res. 2014; 21(3):526-33. PMC: 4315724. DOI: 10.1158/1078-0432.CCR-14-1432. View

Wang R, Zhang Q, Peng X, Zhou C, Zhong Y, Chen X . Stellettin B Induces G1 Arrest, Apoptosis and Autophagy in Human Non-small Cell Lung Cancer A549 Cells via Blocking PI3K/Akt/mTOR Pathway. Sci Rep. 2016; 6:27071. PMC: 4886687. DOI: 10.1038/srep27071. View

10.

Thompson L . Update From the 4th Edition of the World Health Organization Classification of Head and Neck Tumours: Tumours of the Ear. Head Neck Pathol. 2017; 11(1):78-87. PMC: 5340731. DOI: 10.1007/s12105-017-0790-5. View

11.

Makita K, Hamamoto Y, Takata N, Ishikawa H, Tsuruoka S, Uwatsu K . Prognostic significance of inflammatory response markers for locally advanced squamous cell carcinoma of the external auditory canal and middle ear. J Radiat Res. 2021; 62(4):662-668. PMC: 8273788. DOI: 10.1093/jrr/rrab048. View

12.

Schneider C, Rasband W, Eliceiri K . NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012; 9(7):671-5. PMC: 5554542. DOI: 10.1038/nmeth.2089. View

13.

Swaney D, Ramms D, Wang Z, Park J, Goto Y, Soucheray M . A protein network map of head and neck cancer reveals PIK3CA mutant drug sensitivity. Science. 2021; 374(6563):eabf2911. PMC: 9005332. DOI: 10.1126/science.abf2911. View

14.

Brant J, Eliades S, Chen J, Newman J, Ruckenstein M . Carcinoma of the Middle Ear: A Review of the National Cancer Database. Otol Neurotol. 2017; 38(8):1153-1157. DOI: 10.1097/MAO.0000000000001491. View

15.

Lu H, Lu Y, Xie Y, Qiu S, Li X, Fan Z . Rational combination with PDK1 inhibition overcomes cetuximab resistance in head and neck squamous cell carcinoma. JCI Insight. 2019; 4(19). PMC: 6795401. DOI: 10.1172/jci.insight.131106. View

16.

Schafer M, Werner S . Cancer as an overhealing wound: an old hypothesis revisited. Nat Rev Mol Cell Biol. 2008; 9(8):628-38. DOI: 10.1038/nrm2455. View

17.

Nabuurs C, Kievit W, Labbe N, Leemans C, Smit C, van den Brekel M . Evaluation of the modified Pittsburgh classification for predicting the disease-free survival outcome of squamous cell carcinoma of the external auditory canal. Head Neck. 2020; 42(12):3609-3622. PMC: 7754129. DOI: 10.1002/hed.26424. View

18.

Basura G, Smith J, Ellsperman S, Bhangale A, Brenner J . Targeted molecular characterization of external auditory canal squamous cell carcinomas. Laryngoscope Investig Otolaryngol. 2021; 6(5):1151-1157. PMC: 8513438. DOI: 10.1002/lio2.654. View

19.

Yamamoto-Fukuda T, Akiyama N, Kojima H . Super-enhancer Acquisition Drives FOXC2 Expression in Middle Ear Cholesteatoma. J Assoc Res Otolaryngol. 2021; 22(4):405-424. PMC: 8329101. DOI: 10.1007/s10162-021-00801-7. View

20.

Akiyama N, Yamamoto-Fukuda T, Yoshikawa M, Kojima H . Evaluation of YAP signaling in a rat tympanic membrane under a continuous negative pressure load and in human middle ear cholesteatoma. Acta Otolaryngol. 2017; 137(11):1158-1165. DOI: 10.1080/00016489.2017.1351040. View