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

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 expression of canonical markers, DLK1 and MRP proteins than TT cells (p.C634W). MZ-CRC-1 cells also formed more spheroids and showed lower dye accumulation (p < 0.0001). Finally, we found that DLK1+ cells (populations of cells expressing DLK1), exhibited significantly higher expression of stemness markers compared to DLK1- cells (populations of cells negative for DLK1). These findings highlight the role of DLK1 in promoting the stemness phenotype and offer new insights into MTC progression and resistance, with potential therapeutic implications.