DLK1 Is Associated with Stemness Phenotype in Medullary Thyroid Carcinoma Cell Lines

Author(s)

da Silva, Danilo Dias ; Araldi, Rodrigo Pinheiro ; Belizario, Mariana Rocha ; Rocha, Welbert Gomes ; Maciel, Rui Monteiro de Barros ; Cerutti, Janete Maria

• Document type: Journal article
• Source: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES; v. 25, n. 22, p. 19-pg., 2024-11-01.

Abstract

Medullary thyroid carcinoma (MTC) is a rare and aggressive tumor, often requiring systemic treatment in advanced or metastatic stages, where drug resistance presents a significant challenge. Given the role of cancer stem cells (CSCs) in cancer recurrence and drug resistance, we aimed to identify CSC subpopulations within two MTC cell lines harboring pathogenic variants in the two most common MEN2-associated codons. We analyzed 15 stemness-associated markers, along with well-established thyroid stem cell markers (CD133, CD44, and ALDH1), a novel candidate (DLK1), and multidrug resistance proteins (MRP1 and MRP3). The ability to efflux the fluorescent dye Hoechst 3342 and form spheroids, representing CSC behavior, was also assessed. MZ-CRC-1 cells (p.M918T) displayed higher expressions of canonical markers, DLK1, and MRP proteins than TT cells (p.C634W). MZ-CRC-1 cells also formed more spheroids and showed less dye accumulation (p < 0.0001). Finally, we observed that DLK1+ cells (those expressing DLK1) in both cell lines exhibited significantly higher levels of stemness markers compared to DLK1- cells (those lacking DLK1 expression). These findings underscore DLK1's role in enhancing the stemness phenotype, providing valuable insights into MTC progression and resistance and suggesting potential therapeutic implications. (AU)

• FAPESP's process: 23/16471-4 - DLK1 gene editing with CRISPR/Cas9 in cell lines derived from human medullary and differentiated thyroid carcinoma
• Grantee: Mariana Rocha Belizario
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 21/02752-6 - Multiple Endocrine Neoplasia type 2 (MEN 2) and Medullary Thyroid Carcinoma (TCM): new issues in developmental biology, genetics, immunology, epidemiology, mechanisms of disease and clinical management
• Grantee: Rui Monteiro de Barros Maciel
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 22/16827-0 - Evaluation of cooperation between variants in the RAS and HIPK2 genes in the genesis and progression of differentiated thyroid Cancer
• Grantee: Welbert Gomes Rocha
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 22/09713-9 - Identification of predisposing genes to development of Familial Non-Medullary Thryoid Cancer using whole exoma sequencing
• Grantee: Janete Maria Cerutti
• Support Opportunities: Regular Research Grants