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However, the lack of commercially available cSCC cell lines hinders our understanding of this disease. This study aims to establish and characterize a new metastatic cSCC cell line derived from a Brazilian patient. A tumor biopsy was taken from a metastatic cSCC patient, immortalized, and named HCB-541 after several passages. The cytokeratin expression profile, karyotypic alterations, mutational analysis, mRNA and protein differential expression, tumorigenic capacity in xenograft models, and drug sensitivity were analyzed. The HCB-541 cell line showed a doubling time between 20 and 30\u00a0h and high tumorigenic capacity in the xenograft mouse model. The HCB-541 cell line showed hypodiploid and hypotetraploidy populations. We found pathogenic mutations in <jats:italic>TP53<\/jats:italic> p.(Arg248Leu), <jats:italic>HRAS<\/jats:italic> (Gln61His) and <jats:italic>TERT<\/jats:italic> promoter (C228T) and high-level microsatellite instability (MSI-H) in both tumor and cell line. We observed 37 cancer-related genes differentially expressed when compared with HACAT control cells. The HCB-541 cells exhibited high phosphorylated levels of EGFR, AXL, Tie, FGFR, and ROR2, and high sensitivity to cisplatin, carboplatin, and EGFR inhibitors. Our study successfully established HCB-541, a new cSCC cell line that could be useful as a valuable biological model for understanding the biology and therapy of metastatic skin cancer.<\/jats:p>","DOI":"10.1007\/s13577-024-01054-1","type":"journal-article","created":{"date-parts":[[2024,4,3]],"date-time":"2024-04-03T03:41:46Z","timestamp":1712115706000},"page":"1170-1183","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Establishment and molecular characterization of HCB-541, a novel and aggressive human cutaneous squamous cell carcinoma cell line"],"prefix":"10.1007","volume":"37","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4748-8819","authenticated-orcid":false,"given":"Ana Carolina","family":"Laus","sequence":"first","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0551-138X","authenticated-orcid":false,"given":"Izabela Natalia Faria","family":"Gomes","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3260-0847","authenticated-orcid":false,"given":"Aline Larissa Virginio","family":"da Silva","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7470-4655","authenticated-orcid":false,"given":"Luciane Sussuchi","family":"da Silva","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1622-9606","authenticated-orcid":false,"given":"Mirella Baroni","family":"Milan","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2462-8535","authenticated-orcid":false,"given":"Silvia","family":"AparecidaTeixeira","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5426-2223","authenticated-orcid":false,"given":"Ana Carolina Baptista Moreno","family":"Martin","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0874-6600","authenticated-orcid":false,"given":"Let\u00edcia","family":"do Nascimento Braga Pereira","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0009-0005-6041-8307","authenticated-orcid":false,"given":"Carlos Eduardo Barbosa","family":"de Carvalho","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3604-7033","authenticated-orcid":false,"given":"Camila Souza","family":"Crovador","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0009-0001-9236-990X","authenticated-orcid":false,"given":"Fl\u00e1via Escremin","family":"de Paula","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8717-8968","authenticated-orcid":false,"given":"Fl\u00e1via Caroline","family":"Nascimento","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5885-5322","authenticated-orcid":false,"given":"Helder Teixeira","family":"de Freitas","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0325-5514","authenticated-orcid":false,"given":"Vinicius","family":"de Lima Vazquez","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9639-7940","authenticated-orcid":false,"given":"Rui Manuel","family":"Reis","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6500-4190","authenticated-orcid":false,"given":"Renato Jos\u00e9","family":"da Silva-Oliveira","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2024,4,3]]},"reference":[{"key":"1054_CR1","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1016\/j.ejca.2020.01.007","volume":"128","author":"AJ Stratigos","year":"2020","unstructured":"Stratigos AJ, Garbe C, Dessinioti C, Lebbe C, Bataille V, Bastholt L, et al. 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