{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T13:49:51Z","timestamp":1776952191172,"version":"3.51.4"},"reference-count":70,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,1,4]],"date-time":"2022-01-04T00:00:00Z","timestamp":1641254400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation","doi-asserted-by":"publisher","award":["2017\/22305-9"],"award-info":[{"award-number":["2017\/22305-9"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfeicoamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["88887.137283\/2017-00"],"award-info":[{"award-number":["88887.137283\/2017-00"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Cetuximab is the sole anti-EGFR monoclonal antibody that is FDA approved to treat head and neck squamous cell carcinoma (HNSCC). However, no predictive biomarkers of cetuximab response are known for HNSCC. Herein, we address the molecular mechanisms underlying cetuximab resistance in an in vitro model. We established a cetuximab resistant model (FaDu), using increased cetuximab concentrations for more than eight months. The resistance and parental cells were evaluated for cell viability and functional assays. Protein expression was analyzed by Western blot and human cell surface panel by lyoplate. The mutational profile and copy number alterations (CNA) were analyzed using whole-exome sequencing (WES) and the NanoString platform. FaDu resistant clones exhibited at least two-fold higher IC50 compared to the parental cell line. WES showed relevant mutations in several cancer-related genes, and the comparative mRNA expression analysis showed 36 differentially expressed genes associated with EGFR tyrosine kinase inhibitors resistance, RAS, MAPK, and mTOR signaling. Importantly, we observed that overexpression of KRAS, RhoA, and CD44 was associated with cetuximab resistance. Protein analysis revealed EGFR phosphorylation inhibition and mTOR increase in resistant cells. Moreover, the resistant cell line demonstrated an aggressive phenotype with a significant increase in adhesion, the number of colonies, and migration rates. Overall, we identified several molecular alterations in the cetuximab resistant cell line that may constitute novel biomarkers of cetuximab response such as mTOR and RhoA overexpression. These findings indicate new strategies to overcome anti-EGFR resistance in HNSCC.<\/jats:p>","DOI":"10.3390\/cells11010154","type":"journal-article","created":{"date-parts":[[2022,1,5]],"date-time":"2022-01-05T20:42:16Z","timestamp":1641415336000},"page":"154","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Comprehensive Molecular Landscape of Cetuximab Resistance in Head and Neck Cancer Cell Lines"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0551-138X","authenticated-orcid":false,"given":"Izabela N. F.","family":"Gomes","sequence":"first","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"given":"Renato J.","family":"da Silva-Oliveira","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"},{"name":"Barretos School of Medicine Dr. Paulo Prata\u2014FACISB, Barretos 14785-002, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7470-4655","authenticated-orcid":false,"given":"Luciane Sussuchi","family":"da Silva","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"given":"Olga","family":"Martinho","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), Medical School, University of Minho, 4710-057 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4731-2082","authenticated-orcid":false,"given":"Adriane F.","family":"Evangelista","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"given":"Andr\u00e9","family":"van Helvoort Lengert","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2200-3985","authenticated-orcid":false,"given":"Let\u00edcia Ferro","family":"Leal","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"},{"name":"Barretos School of Medicine Dr. Paulo Prata\u2014FACISB, Barretos 14785-002, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5919-5242","authenticated-orcid":false,"given":"Viviane Aline Oliveira","family":"Silva","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"given":"St\u00e9phanie Piancenti","family":"dos Santos","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Diagnosis, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"given":"Fl\u00e1via Caroline","family":"Nascimento","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), Medical School, University of Minho, 4710-057 Braga, Portugal"}]},{"given":"Andr\u00e9","family":"Lopes Carvalho","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9639-7940","authenticated-orcid":false,"given":"Rui Manuel","family":"Reis","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil"},{"name":"Life and Health Sciences Research Institute (ICVS), Medical School, University of Minho, 4710-057 Braga, Portugal"},{"name":"Laboratory of Molecular Diagnosis, Barretos Cancer Hospital, Barretos 14784-400, Brazil"},{"name":"3ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. 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