IFITM3 knockdown reduces the expression of CCND1 and CDK4 and suppresses the growth of oral squamous cell carcinoma cells
- 96 Downloads
Oral squamous cell carcinoma (OSCC) is a challenging disease to treat. Up to 50% of OSCC patients with advanced disease develop recurrences. Elucidation of key molecular mechanisms underlying OSCC development may provide opportunities to target specific genes and, thus, to improve patient survival. In this study, we examined the expression and functional role of interferon transmembrane protein 3 (IFITM3) in OSCC development.
The expression of IFITM3 in OSCC and normal oral mucosal tissues was assessed by qRT-PCR and immunohistochemistry. The role of IFITM3 in driving OSCC cell proliferation and survival was examined using siRNA-mediated gene knockdown, and the role of IFITM3 in driving cell cycle regulators was examined using Western blotting.
We found that IFITM3 is overexpressed in more than 79% of primary OSCCs. We also found that IFITM3 knockdown led to impaired OSCC cell growth through inhibition of cell proliferation, induction of cell cycle arrest, senescence and apoptosis. In addition, we found that IFITM3 knockdown led to reduced expressions of CCND1 and CDK4 and reduced RB phosphorylation, leading to inhibition of OSCC cell growth. This information may be instrumental for the design of novel targeted therapeutic strategies.
From our data we conclude that IFITM3 is overexpressed in OSCC and may regulate the CCND1-CDK4/6-pRB axis to mediate OSCC cell growth.
KeywordsOral squamous cell carcinoma IFITM3 Proliferation CCND1 CDK4
We would like to thank all consented patients who participated in this study. This study was funded by Ministry of Science, Technology and Innovation (MOSTI 06-00-00-0000), University of Malaya (UM.C/625/1/HIR/MOHE/DENT-03) and sponsors of Cancer Research Malaysia. Cancer Research Malaysia is a non-profit research organization. We are committed to an understanding of cancer prevention, diagnosis and treatment through a fundamental research program.
Compliance with ethical standards
Conflict of interest
All authors declared that they have no conflict of interest.
- 3.A.R. Everitt, S. Clare, T. Pertel, S.P. John, R.S. Wash, S.E. Smith, C.R. Chin, E.M. Feeley, J.S. Sims, D.J. Adams, H.M. Wise, L. Kane, D. Goulding, P. Digard, V. Anttila, J.K. Baillie, T.S. Walsh, D.A. Hume, A. Palotie, Y. Xue, V. Colonna, C. Tyler-Smith, J. Dunning, S.B. Gordon, R.L. Smyth, P.J. Openshaw, G. Dougan, A.L. Brass, P. Kellam, IFITM3 restricts the morbidity and mortality associated with influenza. Nature 484, 519–523 (2012)CrossRefGoogle Scholar
- 4.A.L. Brass, I.C. Huang, Y. Benita, S.P. John, M.N. Krishnan, E.M. Feeley, B.J. Ryan, J.L. Weyer, L. van der Weyden, E. Fikrig, D.J. Adams, R.J. Xavier, M. Farzan, S.J. Elledge, The IFITM proteins mediate cellular resistance to influenza A H1N1 virus, West Nile virus, and dengue virus. Cell 139, 1243–1254 (2009)CrossRefGoogle Scholar
- 5.S.S. Evans, R.P. Collea, J.A. Leasure, D.B. Lee, IFN-alpha induces homotypic adhesion and Leu-13 expression in human B lymphoid cells. J Immunol 150, 736–747 (1993)Google Scholar
- 15.A.J. Lui, E.S. Geanes, J. Ogony, F. Behbod, J. Marquess, K. Valdez, W. Jewell, O. Tawfik, J. Lewis-Wambi, IFITM1 suppression blocks proliferation and invasion of aromatase inhibitor-resistant breast cancer in vivo by JAK/STAT-mediated induction of p21. Cancer Lett 399, 29–43 (2017)CrossRefGoogle Scholar
- 20.J. Fan, Z. Peng, C. Zhou, G. Qiu, H. Tang, Y. Sun, X. Wang, Q. Li, X. Le, K. Xie, Gene-expression profiling in Chinese patients with colon cancer by coupling experimental and bioinformatic genomewide gene-expression analyses: Identification and validation of IFITM3 as a biomarker of early colon carcinogenesis. Cancer 113, 266–275 (2008)CrossRefGoogle Scholar
- 28.T. Hisamatsu, M. Watanabe, H. Ogata, T. Ezaki, S. Hozawa, H. Ishii, T. Kanai, T. Hibi, Interferon-inducible gene family 1-8U expression in colitis-associated colon cancer and severely inflamed mucosa in ulcerative colitis. Cancer Res 59, 5927–5931 (1999)Google Scholar
- 29.F. Wu, T. Dassopoulos, L. Cope, A. Maitra, S.R. Brant, M.L. Harris, T.M. Bayless, G. Parmigiani, S. Chakravarti, Genome-wide gene expression differences in Crohn's disease and ulcerative colitis from endoscopic pinch biopsies: Insights into distinctive pathogenesis. Inflamm Bowel Dis 13, 807–821 (2007)CrossRefGoogle Scholar
- 35.R.B. Zain, W.M. Ghani, I.A. Razak, R.J. Latifah, A.R. Samsuddin, S.C. Cheong, N. Abdullah, A.R. Ismail, H.B. Hussaini, N.A. Talib, A. Jallaludin, Building partnership in oral cancer research in a developing country: Processes and barriers. Asian Pac J Cancer Prev 10, 513–518 (2009)Google Scholar
- 38.S. Hamid, K.P. Lim, R.B. Zain, S.M. Ismail, S.H. Lau, W.M. Mustafa, M.T. Abraham, N.A. Nam, S.H. Teo, S.C. Cheong, Establishment and characterization of Asian oral cancer cell lines as in vitro models to study a disease prevalent in Asia. Int J Mol Med 19, 453–460 (2007)Google Scholar
- 39.M.Z. Fadlullah, I.K. Chiang, K.R. Dionne, P.S. Yee, C.P. Gan, K.K. Sam, K.H. Tiong, A.K. Ng, D. Martin, K.P. Lim, T.G. Kallarakkal, W.M. Mustafa, S.H. Lau, M.T. Abraham, R.B. Zain, Z.A. Rahman, A. Molinolo, V. Patel, J.S. Gutkind, A.C. Tan, S.C. Cheong, Genetically-defined novel oral squamous cell carcinoma cell lines for the development of molecular therapies. Oncotarget 7, 27802–27818 (2016)CrossRefGoogle Scholar
- 41.D. Szklarczyk, J.H. Morris, H. Cook, M. Kuhn, S. Wyder, M. Simonovic, A. Santos, N.T. Doncheva, A. Roth, P. Bork, L.J. Jensen, C. von Mering, The STRING database in 2017: Quality-controlled protein-protein association networks, made broadly accessible. Nucleic Acids Res 45, D362–D368 (2017)CrossRefGoogle Scholar
- 46.K. Kollmann, G. Heller, C. Schneckenleithner, W. Warsch, R. Scheicher, R.G. Ott, M. Schafer, S. Fajmann, M. Schlederer, A.I. Schiefer, U. Reichart, M. Mayerhofer, C. Hoeller, S. Zochbauer-Muller, D. Kerjaschki, C. Bock, L. Kenner, G. Hoefler, M. Freissmuth, A.R. Green, R. Moriggl, M. Busslinger, M. Malumbres, V. Sexl, A kinase-independent function of CDK6 links the cell cycle to tumor angiogenesis. Cancer Cell 30, 359–360 (2016)CrossRefGoogle Scholar
- 47.J.J. Lucas, J. Domenico, E.W. Gelfand, Cyclin-dependent kinase 6 inhibits proliferation of human mammary epithelial cells. Mol Cancer Res 2, 105–114 (2004)Google Scholar
- 48.C.R. Pickering, J. Zhang, S.Y. Yoo, L. Bengtsson, S. Moorthy, D.M. Neskey, M. Zhao, M.V. Ortega Alves, K. Chang, J. Drummond, E. Cortez, T.X. Xie, D. Zhang, W. Chung, J.P. Issa, P.A. Zweidler-McKay, X. Wu, A.K. El-Naggar, J.N. Weinstein, J. Wang, D.M. Muzny, R.A. Gibbs, D.A. Wheeler, J.N. Myers, M.J. Frederick, Integrative genomic characterization of oral squamous cell carcinoma identifies frequent somatic drivers. Cancer Discov 3, 770–781 (2013)CrossRefGoogle Scholar
- 49.T.N. Beck, E.A. Golemis, Genomic insights into head and neck cancer. Cancers of the head & neck 1, (2016). https://doi.org/10.1186/s41199-016-0003-z
- 50.T.C.G.A. Network, M.S. Lawrence, C. Sougnez, L. Lichtenstein, K. Cibulskis, E. Lander, S.B. Gabriel, G. Getz, A. Ally, M. Balasundaram, I. Birol, R. Bowlby, D. Brooks, Y.S.N. Butterfield, R. Carlsen, D. Cheng, A. Chu, N. Dhalla, R. Guin, R.A. Holt, S.J.M. Jones, D. Lee, H.I. Li, M.A. Marra, M. Mayo, R.A. Moore, A.J. Mungall, A.G. Robertson, J.E. Schein, P. Sipahimalani, A. Tam, N. Thiessen, T. Wong, A. Protopopov, N. Santoso, S. Lee, M. Parfenov, J. Zhang, H.S. Mahadeshwar, J. Tang, X. Ren, S. Seth, P. Haseley, D. Zeng, L. Yang, A.W. Xu, X. Song, A. Pantazi, C.A. Bristow, A. Hadjipanayis, J. Seidman, L. Chin, P.J. Park, R. Kucherlapati, R. Akbani, T. Casasent, W. Liu, Y. Lu, G. Mills, T. Motter, J. Weinstein, L. Diao, J. Wang, Y.H. Fan, J. Liu, K. Wang, J.T. Auman, S. Balu, T. Bodenheimer, E. Buda, D.N. Hayes, K.A. Hoadley, A.P. Hoyle, S.R. Jefferys, C.D. Jones, P.K. Kimes, Y. Liu, J.S. Marron, S. Meng, P.A. Mieczkowski, L.E. Mose, J.S. Parker, C.M. Perou, J.F. Prins, J. Roach, Y. Shi, J.V. Simons, D. Singh, M.G. Soloway, D. Tan, U. Veluvolu, V. Walter, S. Waring, M.D. Wilkerson, J. Wu, N. Zhao, A.D. Cherniack, P.S. Hammerman, A.D. Tward, C.S. Pedamallu, G. Saksena, J. Jung, A.I. Ojesina, S.L. Carter, T.I. Zack, S.E. Schumacher, R. Beroukhim, S.S. Freeman, M. Meyerson, J. Cho, L. Chin, G. Getz, M.S. Noble, D. DiCara, H. Zhang, D.I. Heiman, N. Gehlenborg, D. Voet, P. Lin, S. Frazer, P. Stojanov, Y. Liu, L. Zou, J. Kim, C. Sougnez, S.B. Gabriel, M.S. Lawrence, D. Muzny, H. Doddapaneni, C. Kovar, J. Reid, D. Morton, Y. Han, W. Hale, H. Chao, K. Chang, J.A. Drummond, R.A. Gibbs, N. Kakkar, D. Wheeler, L. Xi, G. Ciriello, M. Ladanyi, W. Lee, R. Ramirez, C. Sander, R. Shen, R. Sinha, N. Weinhold, B.S. Taylor, B.A. Aksoy, G. Dresdner, J. Gao, B. Gross, A. Jacobsen, B. Reva, N. Schultz, S.O. Sumer, Y. Sun, T.A. Chan, L.G. Morris, J. Stuart, S. Benz, S. Ng, C. Benz, C. Yau, S.B. Baylin, L. Cope, L. Danilova, J.G. Herman, M. Bootwalla, D.T. Maglinte, P.W. Laird, T. Triche Jr., D.J. Weisenberger, D.J. Van Den Berg, N. Agrawal, J. Bishop, P.C. Boutros, J.P. Bruce, L.A. Byers, J. Califano, T.E. Carey, Z. Chen, H. Cheng, S.I. Chiosea, E. Cohen, B. Diergaarde, A.M. Egloff, A.K. El-Naggar, R.L. Ferris, M.J. Frederick, J.R. Grandis, Y. Guo, R.I. Haddad, P.S. Hammerman, T. Harris, D.N. Hayes, A.B.Y. Hui, J.J. Lee, S.M. Lippman, F.-F. Liu, J.B. McHugh, J. Myers, P.K.S. Ng, B. Perez-Ordonez, C.R. Pickering, M. Prystowsky, M. Romkes, A.D. Saleh, M.A. Sartor, R. Seethala, T.Y. Seiwert, H. Si, A.D. Tward, C. Van Waes, D.M. Waggott, M. Wiznerowicz, W.G. Yarbrough, J. Zhang, Z. Zuo, K. Burnett, D. Crain, J. Gardner, K. Lau, D. Mallery, S. Morris, J. Paulauskis, R. Penny, C. Shelton, T. Shelton, M. Sherman, P. Yena, A.D. Black, J. Bowen, J. Frick, J.M. Gastier-Foster, H.A. Harper, K. Leraas, T.M. Lichtenberg, N.C. Ramirez, L. Wise, E. Zmuda, J. Baboud, M.A. Jensen, A.B. Kahn, T.D. Pihl, D.A. Pot, D. Srinivasan, J.S. Walton, Y. Wan, R.A. Burton, T. Davidsen, J.A. Demchok, G. Eley, M.L. Ferguson, K.R.M. Shaw, B.A. Ozenberger, M. Sheth, H.J. Sofia, R. Tarnuzzer, Z. Wang, L. Yang, J.C. Zenklusen, C. Saller, K. Tarvin, C. Chen, R. Bollag, P. Weinberger, W. Golusiński, P. Golusiński, M. Ibbs, K. Korski, A. Mackiewicz, W. Suchorska, B. Szybiak, M. Wiznerowicz, K. Burnett, E. Curley, J. Gardner, D. Mallery, R. Penny, T. Shelton, P. Yena, C. Beard, C. Mitchell, G. Sandusky, N. Agrawal, J. Ahn, J. Bishop, J. Califano, Z. Khan, J.P. Bruce, A.B.Y. Hui, J. Irish, F.-F. Liu, B. Perez-Ordonez, J. Waldron, P.C. Boutros, D.M. Waggott, J. Myers, W.N. William Jr., S.M. Lippman, S. Egea, C. Gomez-Fernandez, L. Herbert, C.R. Bradford, T.E. Carey, D.B. Chepeha, A.S. Haddad, T.R. Jones, C.M. Komarck, M. Malakh, J.B. McHugh, J.S. Moyer, A. Nguyen, L.A. Peterson, M.E. Prince, L.S. Rozek, M.A. Sartor, E.G. Taylor, H.M. Walline, G.T. Wolf, L. Boice, B.S. Chera, W.K. Funkhouser, M.L. Gulley, T.G. Hackman, D.N. Hayes, M.C. Hayward, M. Huang, W.K. Rathmell, A.H. Salazar, W.W. Shockley, C.G. Shores, L. Thorne, M.C. Weissler, S. Wrenn, A.M. Zanation, S.I. Chiosea, B. Diergaarde, A.M. Egloff, R.L. Ferris, M. Romkes, R. Seethala, B.T. Brown, Y. Guo, M. Pham, W.G. Yarbrough, Comprehensive genomic characterization of head and neck squamous cell carcinomas. Nature 517, 576–582 (2015)CrossRefGoogle Scholar
- 51.L. Michel, J. Ley, T.M. Wildes, A. Schaffer, A. Robinson, S.E. Chun, W. Lee, J. Lewis Jr., K. Trinkaus, D. Adkins, Phase I trial of palbociclib, a selective cyclin dependent kinase 4/6 inhibitor, in combination with cetuximab in patients with recurrent/metastatic head and neck squamous cell carcinoma. Oral Oncol 58, 41–48 (2016)CrossRefGoogle Scholar