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DLK1 Is Associated with Stemness Phenotype in Medullary Thyroid Carcinoma Cell Lines
Danilo Dias da Silva1, Rodrigo Pinheiro Araldi1, Mariana Rocha Belizario1
1Genetic Bases of Thyroid Tumour Laboratory, Division of Genetics, Department of Morphology and Genetics, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo 04039-032, SP, Brazil.
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.
Insights
This study identifies cancer stem cell (CSC) subpopulations in medullary thyroid carcinoma (MTC). DLK1 expression enhances CSC stemness, offering insights into MTC progression and drug resistance.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Medullary thyroid carcinoma (MTC) is aggressive, with drug resistance a major challenge in advanced stages.
- Cancer stem cells (CSCs) are implicated in cancer recurrence and treatment failure.
Purpose of the Study:
- To identify CSC subpopulations in MTC cell lines with common MEN2-associated variants.
- To investigate the role of DLK1 and multidrug resistance proteins (MRPs) in MTC stemness.
Main Methods:
- Analysis of 15 stemness markers, CD133, CD44, ALDH1, DLK1, MRP1, and MRP3 in two MTC cell lines (MZ-CRC-1 and TT).
- Assessment of Hoechst 3342 dye efflux and spheroid formation to evaluate CSC behavior.
- Comparison of stemness marker expression between DLK1-positive and DLK1-negative cells.
Main Results:
- MZ-CRC-1 cells (p.M918T) showed higher expression of stemness markers, DLK1, and MRPs compared to TT cells (p.C634W).
- MZ-CRC-1 cells exhibited enhanced spheroid formation and reduced dye efflux, indicative of greater CSC activity.
- DLK1-positive cells displayed significantly higher stemness marker levels than DLK1-negative cells in both MTC lines.
Conclusions:
- DLK1 plays a crucial role in promoting the stemness phenotype in MTC.
- These findings provide insights into MTC progression and drug resistance mechanisms.
- DLK1 may represent a potential therapeutic target for MTC treatment.
