Anti-EGFR treatment effects on laryngeal cancer stem cells

Author(s):Show less -

de Mendonca Fernandes, Glaucia Maria ^[1] ; Silva Galbiatti-Dias, Ana Livia ^[1] ; Muniz Ferreira, Leticia Antunes ^[1] ; Serafim Junior, Vilson ^[1] ; Rodrigues-Fleming, Gabriela Helena ^[1] ; de Oliveira-Cucolo, Juliana Garcia ^[1] ; Biselli-Chicote, Patricia Matos ^[1] ; Kawasaki-Oyama, Rosa Sayoko ^[1] ; Maniglia, Jose Victor ^[2] ; Pavarino, Erika Cristina ^[1] ; Goloni-Bertollo, Eny Maria ^[1]

Affiliation

^[1] Sao Jose do Rio Preto Med Sch FAMERP, Genet & Mol Biol Res Unit UPGEM, Sao Jose Do Rio Preto, SP - Brazil

^[2] Sao Jose Do Rio Preto Med Sch FAMERP, Dept Otolaryngol Head & Neck Surg, Sao Jose Do Rio Preto, SP - Brazil

• Document type: Journal article
• Source: AMERICAN JOURNAL OF TRANSLATIONAL RESEARCH; v. 13, n. 1, p. 143-155, 2021.
• Web of Science Citations: 0

Abstract

Laryngeal cancer (LC) is one of the common head and neck neoplasms and is characterized by resistance to conventional therapy and poor prognosis. This may result from the presence of cancer stem cells (CSCs), which form a small population in tumors with metastatic potential, high invasive capacity, self-renewal, and differentiation. This study aimed to evaluate the effectiveness of 5-fluorouracil and cisplatin individually, as well as the combination of cetuximab and paclitaxel in a CSC subpopulation separated with biomarkers related to tumoral growth (CD44, CD117, and CD133). In addition, expression of TrkB, KRAS, HIF-1 alpha, and VEGF-A genes and proteins related to cell proliferation were evaluated in this subpopulation. The CD44, CD133, and CD117 biomarkers were used to analyze the identification and separation of both subpopulations using FACSAria Fusion. Subpopulations positive for CD44, CD133, and CD117 or lacking these biomarkers were classified as laryngeal cancer stem cells (LCSCs) or laryngeal cancer non-stem cells (non-LCSCs), respectively. Matrigel invasion and colony forming assays were performed to confirm CSC presence. Subpopulations were cultured and exposed to 5-fluorouracil, cisplatin, and cetuximab/paclitaxel drugs for 24 h. Cell proliferation was determined using MTS assay. KRAS and TrkB gene expression levels were evaluated using quantitative real time PCR with TaqMan (R) Assay in both subpopulations. The non-LCSC subpopulation was considered as the control for relative expression. We found that the LCSC subpopulation demonstrated more resistance to cetuximab and paclitaxel combination chemotherapy when compared with the non-LCSC subpopulation of the cell line. These LCSC subpopulations presented up-regulated expression of KRAS, HIF-1 alpha, and VEGF- A genes and proteins and no TrkB gene expression, but TrkB protein expression was up-regulated in the LC cell line when compared to the non-CSC subpopulation. ``In conclusion, the combination of CD44, CD133, and CD117 biomarkers has stem cell properties. Moreover, LCSCs, are capable of resisting treatment and present high KRAS, HIF-1 alpha, and VEGF- A gene expression{''}. (AU)

• FAPESP's process: 16/20087-1 - Evaluation of miRNAs expression that regulates the ZEB1, involved in epithelial-mesenchymal transition in head and neck cancer stem cells
• Grantee: José Victor Maniglia
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/04403-8 - Evaluation of angiogenesis biomarkers in cancer development and response to treatment.
• Grantee: Eny Maria Goloni Bertollo
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/15009-6 - Identification of tumoral stem cells in head and neck cancer: genetic and protein expression in the response to chemotherapy
• Grantee: Ana Lívia Silva Galbiatti Dias
• Support Opportunities: Scholarships in Brazil - Post-Doctoral