{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"Research Square"}],"indexed":{"date-parts":[[2024,2,22]],"date-time":"2024-02-22T15:57:34Z","timestamp":1708617454840},"posted":{"date-parts":[[2023,4,25]]},"group-title":"In Review","reference-count":36,"publisher":"Research Square Platform LLC","license":[{"start":{"date-parts":[[2023,4,25]],"date-time":"2023-04-25T00:00:00Z","timestamp":1682380800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,4,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n        <jats:p>In the present study, we describe the establishment of a Brazilian primary laryngeal HNSCC cell line, HCB-289, and isolated a putative cancer stem-like cell subpopulation. Primary cell culture was established from a mechanically disrupted fresh tumor by adherence to cell culture flask. Tumor cells were enriched by fibroblast elimination via differential trypsinization. After establishment, a CD44HIGH \/ALDHHIGH subpopulation was isolated and purified from the parental HCB-289 cells by cell sorting. The CD44HIGH \/ALDHHIGH proliferation rates and lower sensitivity to cisplatin, paclitaxel, cetuximab, and allitinib than its CD44LOW \/ALDHLOW counterpart. Moreover, nanoString subpopulation exhibited greater clonogenic potential, lower miRNome analysis of these two subpopulations showed a higher expression of has-miR-205-5p CD44HIGH \/ALDHHIGH and a lower expression of has-miR146a-5p (ratio = 0.08) in the subpopulation (ratio = 35.37). This novel study model will help in the understanding of the biology and molecular features of larynx squamous cell carcinoma, and its chemotherapeutic resistance. \n<jats:bold>Implications:<\/jats:bold>Our study established and characterized a new cell lineage from the Brazilian population derived from a solid primary tumor of the laryngeal HNSCC primary cell line and can be a useful tool for studying laryngeal squamous cell carcinoma<\/jats:p>","DOI":"10.21203\/rs.3.rs-2787430\/v1","type":"posted-content","created":{"date-parts":[[2023,4,25]],"date-time":"2023-04-25T15:58:46Z","timestamp":1682438326000},"source":"Crossref","is-referenced-by-count":1,"title":["Biological and molecular characterization of HCB-289: a Brazilian head and neck cancer stem-like cell line"],"prefix":"10.21203","author":[{"given":"Matias Eliseo","family":"Melendez","sequence":"first","affiliation":[{"name":"Brazilian National Cancer Institute"}]},{"given":"Renato Jos\u00e9","family":"Silva-Oliveira","sequence":"additional","affiliation":[{"name":"Barretos Cancer Hospital"}]},{"given":"Adriane Feij\u00f3","family":"Evangelista","sequence":"additional","affiliation":[{"name":"Barretos Cancer Hospital"}]},{"given":"Ana Rubia Alcantara","family":"Pelloso","sequence":"additional","affiliation":[{"name":"Brazilian National Cancer Institute"}]},{"given":"Lidia Maria Rebolho","family":"Arantes","sequence":"additional","affiliation":[{"name":"Barretos Cancer Hospital"}]},{"given":"Rui Manuel","family":"Reis","sequence":"additional","affiliation":[{"name":"Barretos Cancer Hospital"}]},{"given":"Ana Carolina","family":"De Carvalho","sequence":"additional","affiliation":[{"name":"Barretos Cancer Hospital"}]},{"given":"Andr\u00e9 Lopes","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Barretos Cancer Hospital"}]}],"member":"8761","reference":[{"key":"ref1","doi-asserted-by":"crossref","first-page":"2289","DOI":"10.1016\/S0140-6736(21)01550-6","article-title":"Head and neck cancer","volume":"398","author":"Mody MD","year":"2021","unstructured":"Mody, M. D., Rocco, J. W., Yom, S. S., Haddad, R. I. & Saba, N. F. Head and neck cancer. The Lancet 398, 2289\u20132299 (2021).","journal-title":"The Lancet"},{"key":"ref2","doi-asserted-by":"crossref","first-page":"394","DOI":"10.3322\/caac.21492","article-title":"Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"68","author":"Bray F","year":"2018","unstructured":"Bray, F. et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68, 394\u2013424 (2018).","journal-title":"CA Cancer J Clin"},{"key":"ref3","unstructured":"INCA. Estimativa 2020: Incid\u00eancia de c\u00e2ncer no Brasil. https:\/\/www.inca.gov.br\/estimativa\/introducao (2020)."},{"key":"ref4","doi-asserted-by":"crossref","DOI":"10.1038\/s41572-020-00224-3","article-title":"Head and neck squamous cell carcinoma","volume":"6","author":"Johnson DE","year":"2020","unstructured":"Johnson, D. E. et al. Head and neck squamous cell carcinoma. Nat Rev Dis Primers 6, (2020).","journal-title":"Nat Rev Dis Primers"},{"key":"ref5","doi-asserted-by":"crossref","first-page":"1695","DOI":"10.1016\/S0140-6736(08)60728-X","article-title":"Head and neck cancer","volume":"371","author":"Argiris A","year":"2008","unstructured":"Argiris, A., Karamouzis, M. V., Raben, D. & Ferris, R. L. Head and neck cancer. Lancet 371, 1695\u20131709 (2008).","journal-title":"Lancet"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1158\/1078-0432.CCR-07-0722","article-title":"Evaluation of Promoter Hypermethylation Detection in Body Fluids as a Screening\/Diagnosis Tool for Head and Neck Squamous Cell Carcinoma","volume":"14","author":"Carvalho AL","year":"2008","unstructured":"Carvalho, A. L. et al. Evaluation of Promoter Hypermethylation Detection in Body Fluids as a Screening\/Diagnosis Tool for Head and Neck Squamous Cell Carcinoma. Clinical Cancer Research 14, 97\u2013107 (2008).","journal-title":"Clinical Cancer Research"},{"key":"ref7","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1016\/0360-3016(94)90306-9","article-title":"The effect of loco-regional control on distant metastatic dissemination in carcinoma of the nasopharynx: An analysis of 1301 patients","volume":"30","author":"Kwong D","year":"1994","unstructured":"Kwong, D., Sham, J. & Choy, D. The effect of loco-regional control on distant metastatic dissemination in carcinoma of the nasopharynx: An analysis of 1301 patients. Int J Radiat Oncol Biol Phys 30, 1029\u20131036 (1994).","journal-title":"Int J Radiat Oncol Biol Phys"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1056\/NEJMra1715715","article-title":"Head and Neck Cancer","volume":"382","author":"Longo DL","year":"2020","unstructured":"Longo, D. L. & Chow, L. Q. M. Head and Neck Cancer. New England Journal of Medicine 382, 60\u201372 (2020).","journal-title":"New England Journal of Medicine"},{"key":"ref9","author":"Lawrence MS","year":"2015","unstructured":"Lawrence, M. S. et al. Comprehensive genomic characterization of head and neck squamous cell carcinomas. Nature 2015 517:7536 517, 576\u2013582 (2015)."},{"key":"ref10","author":"Leemans CR","year":"2018","unstructured":"Leemans, C. R., Snijders, P. J. F. & Brakenhoff, R. H. The molecular landscape of head and neck cancer. Nature Reviews Cancer 2018 18:5 18, 269\u2013282 (2018)."},{"key":"ref11","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/j.ijbiomac.2020.01.131","article-title":"Inhibition of CD44 sensitizes cisplatin-resistance and affects Wnt\/\u03b2-catenin signaling in HNSCC cells","volume":"149","author":"Roy S","year":"2020","unstructured":"Roy, S. et al. Inhibition of CD44 sensitizes cisplatin-resistance and affects Wnt\/\u03b2-catenin signaling in HNSCC cells. Int J Biol Macromol 149, 501\u2013512 (2020).","journal-title":"Int J Biol Macromol"},{"key":"ref12","author":"Koo BS","year":"2014","unstructured":"Koo, B. S. et al. Oct4 is a critical regulator of stemness in head and neck squamous carcinoma cells. Oncogene 2015 34:18 34, 2317\u20132324 (2014)."},{"key":"ref13","doi-asserted-by":"crossref","first-page":"3603","DOI":"10.1007\/s13277-013-0941-6","article-title":"Clinical implications of cancer stem cell-like side population cells in human laryngeal cancer","volume":"34","author":"Yu D","year":"2013","unstructured":"Yu, D. et al. Clinical implications of cancer stem cell-like side population cells in human laryngeal cancer. Tumour Biol 34, 3603\u20133610 (2013).","journal-title":"Tumour Biol"},{"key":"ref14","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1073\/pnas.0610117104","article-title":"Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma","volume":"104","author":"Prince ME","year":"2007","unstructured":"Prince, M. E. et al. Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci U S A 104, 973\u2013978 (2007).","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref15","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1007\/s00432-014-1685-4","article-title":"Prognostic significance of ALDH1A1-positive cancer stem cells in patients with locally advanced, metastasized head and neck squamous cell carcinoma","volume":"140","author":"Qian X","year":"2014","unstructured":"Qian, X. et al. Prognostic significance of ALDH1A1-positive cancer stem cells in patients with locally advanced, metastasized head and neck squamous cell carcinoma. J Cancer Res Clin Oncol 140, 1151\u20131158 (2014).","journal-title":"J Cancer Res Clin Oncol"},{"key":"ref16","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.3892\/or.2013.2340","article-title":"ALDH1-positive cancer stem-like cells are enriched in nodal metastases of oropharyngeal squamous cell carcinoma independent of HPV status","volume":"29","author":"Qian X","year":"2013","unstructured":"Qian, X. et al. ALDH1-positive cancer stem-like cells are enriched in nodal metastases of oropharyngeal squamous cell carcinoma independent of HPV status. Oncol Rep 29, 1777\u20131784 (2013).","journal-title":"Oncol Rep"},{"key":"ref17","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1007\/s13402-016-0270-z","article-title":"Cytotoxicity of allitinib, an irreversible anti-EGFR agent, in a large panel of human cancer-derived cell lines: KRAS mutation status as a predictive biomarker","volume":"39","author":"Silva-Oliveira RJ","year":"2016","unstructured":"Silva-Oliveira, R. J. et al. Cytotoxicity of allitinib, an irreversible anti-EGFR agent, in a large panel of human cancer-derived cell lines: KRAS mutation status as a predictive biomarker. Cell Oncol (Dordr) 39, 253\u2013263 (2016).","journal-title":"Cell Oncol (Dordr)"},{"key":"ref18","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1007\/978-1-61779-080-5_5","article-title":"Online verification of human cell line identity by STR DNA typing","volume":"731","author":"Dirks WG","year":"2011","unstructured":"Dirks, W. G. & Drexler, H. G. Online verification of human cell line identity by STR DNA typing. Methods Mol Biol 731, 45\u201355 (2011).","journal-title":"Methods Mol Biol"},{"key":"ref19","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/978-1-61779-080-5_14","article-title":"Isolation and culture of squamous cell carcinoma lines","volume":"731","author":"Purdie KJ","year":"2011","unstructured":"Purdie, K. J., Pourreyron, C. & South, A. P. Isolation and culture of squamous cell carcinoma lines. Methods Mol Biol 731, 151\u2013159 (2011).","journal-title":"Methods Mol Biol"},{"key":"ref20","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1038\/s41598-018-38315-7","article-title":"Establishment, molecular and biological characterization of HCB-514: a novel human cervical cancer cell line","volume":"9","author":"Rosa MN","year":"2019","unstructured":"Rosa, M. N. et al. Establishment, molecular and biological characterization of HCB-514: a novel human cervical cancer cell line. Sci Rep 9, 1913 (2019).","journal-title":"Sci Rep"},{"key":"ref21","author":"Souza KCB","year":"2019","unstructured":"Souza, K. C. B. et al. Identification of Cell-Free Circulating MicroRNAs for the Detection of Early Breast Cancer and Molecular Subtyping. J Oncol 2019, (2019)."},{"key":"ref22","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1007\/s12253-018-0462-0","article-title":"Clinical and Molecular Characterization of Surgically Treated Oropharynx Squamous Cell Carcinoma Samples","volume":"25","author":"Carvalho AC","year":"2019","unstructured":"de Carvalho, A. C. et al. Clinical and Molecular Characterization of Surgically Treated Oropharynx Squamous Cell Carcinoma Samples. Pathol Oncol Res 25, 1047\u20131058 (2019).","journal-title":"Pathol Oncol Res"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1016\/j.ijrobp.2014.04.044","article-title":"Human papillomavirus (HPV) infection in squamous cell carcinomas arising from the oropharynx: detection of HPV DNA and p16 immunohistochemistry as diagnostic and prognostic indicators\u2013a pilot study","volume":"89","author":"Bussu F","year":"2014","unstructured":"Bussu, F. et al. Human papillomavirus (HPV) infection in squamous cell carcinomas arising from the oropharynx: detection of HPV DNA and p16 immunohistochemistry as diagnostic and prognostic indicators\u2013a pilot study. Int J Radiat Oncol Biol Phys 89, 1115\u20131120 (2014).","journal-title":"Int J Radiat Oncol Biol Phys"},{"key":"ref24","first-page":"180005","article-title":"Fraction of head and neck cancer attributable to tobacco and alcohol in cities of three Brazilian regions","volume":"21","author":"Kfouri SA","year":"2018","unstructured":"Kfouri, S. A. et al. Fraction of head and neck cancer attributable to tobacco and alcohol in cities of three Brazilian regions. Revista Brasileira de Epidemiologia 21, 180005 (2018).","journal-title":"Revista Brasileira de Epidemiologia"},{"key":"ref25","doi-asserted-by":"crossref","first-page":"7656","DOI":"10.1073\/pnas.141237598","article-title":"Cell culture forensics","volume":"98","author":"O\u2019Brien SJ","year":"2001","unstructured":"O\u2019Brien, S. J. Cell culture forensics. Proc Natl Acad Sci U S A 98, 7656 (2001).","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"Suppl 1","key":"ref26","doi-asserted-by":"crossref","first-page":"E970","DOI":"10.1002\/hed.24137","article-title":"Efficacy and safety of a cisplatin and paclitaxel induction regimen followed by chemoradiotherapy for patients with locally advanced head and neck squamous cell carcinoma","volume":"38","author":"Viana Souza","year":"2016","unstructured":"De Souza Viana, L. et al. Efficacy and safety of a cisplatin and paclitaxel induction regimen followed by chemoradiotherapy for patients with locally advanced head and neck squamous cell carcinoma. Head Neck 38 Suppl 1, E970\u2013E980 (2016).","journal-title":"Head Neck"},{"key":"ref27","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1111\/jop.12348","article-title":"CD44(high) \/ALDH1(high) head and neck squamous cell carcinoma cells exhibit mesenchymal characteristics and GSK3\u03b2-dependent cancer stem cell properties","volume":"45","author":"Seino S","year":"2016","unstructured":"Seino, S. et al. CD44(high) \/ALDH1(high) head and neck squamous cell carcinoma cells exhibit mesenchymal characteristics and GSK3\u03b2-dependent cancer stem cell properties. J Oral Pathol Med 45, 180\u2013188 (2016).","journal-title":"J Oral Pathol Med"},{"key":"ref28","doi-asserted-by":"crossref","first-page":"2013","DOI":"10.7314\/APJCP.2014.15.5.2013","article-title":"ALDH1 in combination with CD44 as putative cancer stem cell markers are correlated with poor prognosis in urothelial carcinoma of the urinary bladder","volume":"15","author":"Keymoosi H","year":"2014","unstructured":"Keymoosi, H., Gheytanchi, E., Asgari, M., Shariftabrizi, A. & Madjd, Z. ALDH1 in combination with CD44 as putative cancer stem cell markers are correlated with poor prognosis in urothelial carcinoma of the urinary bladder. Asian Pac J Cancer Prev 15, 2013\u20132020 (2014).","journal-title":"Asian Pac J Cancer Prev"},{"key":"ref29","author":"Torrezan GT","year":"2014","unstructured":"Torrezan, G. T. et al. Recurrent somatic mutation in DROSHA induces microRNA profile changes in Wilms tumour. Nature Communications 2014 5:1 5, 1\u201310 (2014)."},{"key":"ref30","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1016\/j.ajpath.2011.11.015","article-title":"ALDH1-bright epithelial ovarian cancer cells are associated with CD44 expression, drug resistance, and poor clinical outcome","volume":"180","author":"Wang YC","year":"2012","unstructured":"Wang, Y. C. et al. ALDH1-bright epithelial ovarian cancer cells are associated with CD44 expression, drug resistance, and poor clinical outcome. Am J Pathol 180, 1159\u20131169 (2012).","journal-title":"Am J Pathol"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1038\/nrc3184","article-title":"Cancer stem cells: an evolving concept","volume":"12","author":"Nguyen LV","year":"2012","unstructured":"Nguyen, L. V., Vanner, R., Dirks, P. & Eaves, C. J. Cancer stem cells: an evolving concept. Nat Rev Cancer 12, 133\u2013143 (2012).","journal-title":"Nat Rev Cancer"},{"key":"ref32","doi-asserted-by":"crossref","first-page":"7252","DOI":"10.1158\/1078-0432.CCR-06-0626","article-title":"Mechanisms of egfr gene transcription modulation: relationship to cancer risk and therapy response","volume":"12","author":"Brandt B","year":"2006","unstructured":"Brandt, B., Meyer-Staeckling, S., Schmidt, H., Agelopoulos, K. & Buerger, H. Mechanisms of egfr gene transcription modulation: relationship to cancer risk and therapy response. Clin Cancer Res 12, 7252\u20137260 (2006).","journal-title":"Clin Cancer Res"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"55","DOI":"10.3233\/CBM-150540","article-title":"Unpredictable changes of selected miRNA in expression profile of HNSCC","volume":"16","author":"Lamperska KM","year":"2016","unstructured":"Lamperska, K. M. et al. Unpredictable changes of selected miRNA in expression profile of HNSCC. Cancer Biomark 16, 55\u201364 (2016).","journal-title":"Cancer Biomark"},{"key":"ref34","doi-asserted-by":"crossref","DOI":"10.3390\/cancers11091347","article-title":"The miR-205-5p\/BRCA1\/RAD17 Axis Promotes Genomic Instability in Head and Neck Squamous Cell Carcinomas","volume":"11","author":"Valenti F","year":"2019","unstructured":"Valenti, F. et al. The miR-205-5p\/BRCA1\/RAD17 Axis Promotes Genomic Instability in Head and Neck Squamous Cell Carcinomas. Cancers (Basel) 11, (2019).","journal-title":"Cancers (Basel)"},{"key":"ref35","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/j.humpath.2010.07.020","article-title":"Down-regulation of microRNAs of the miR-200 family and miR-205, and an altered expression of classic and desmosomal cadherins in spindle cell carcinoma of the head and neck\u2013hallmark of epithelial-mesenchymal transition","volume":"42","author":"Zidar N","year":"2011","unstructured":"Zidar, N. et al. Down-regulation of microRNAs of the miR-200 family and miR-205, and an altered expression of classic and desmosomal cadherins in spindle cell carcinoma of the head and neck\u2013hallmark of epithelial-mesenchymal transition. Hum Pathol 42, 482\u2013488 (2011).","journal-title":"Hum Pathol"},{"key":"ref36","author":"ADDITIONAL","unstructured":"ADDITIONAL INFORMATION"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/www.researchsquare.com\/article\/rs-2787430\/v1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.researchsquare.com\/article\/rs-2787430\/v1.html","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,21]],"date-time":"2023-07-21T07:32:30Z","timestamp":1689924750000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.researchsquare.com\/article\/rs-2787430\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,25]]},"references-count":36,"URL":"https:\/\/doi.org\/10.21203\/rs.3.rs-2787430\/v1","relation":{},"subject":[],"published":{"date-parts":[[2023,4,25]]},"subtype":"preprint"}}