Modulating cancer stem cell characteristics in CD133+ melanoma cells through HIF1α, KLF4, and SHH silencing

Malignant melanoma, an aggressive skin cancer derived from melanocytes, contains a subpopulation known as cancer stem cells (CSCs), with distinct self-renewal and differentiation abilities, setting them apart from non-cancer stem cells (NCSCs). This study aims to examine how CSCs respond to the suppression of their stem cell characteristics through targeted gene silencing of HIF1α, KLF4, and SHH within the context of the extracellular matrix, with a particular focus on Matrigel. Silencing targeted genes individually induced distinct changes in CSCs behavior, revealing novel therapeutic targets through analysis of gene expression, protein levels, and cell cycle dynamics. A comparison between melanoma CSCs and NCSCs revealed significant shifts in SHH signaling, epigenetic markers, differentiation, migration, and vascularization genes. Specifically, CSCs exhibited elevated levels of SHH, Gli1, and HDAC9, while NCSCs showed increased expression of Mif. Our findings highlight the emergence of a unique cellular phenotype following gene silencing, distinct from both CSCs and NCSCs. Diverse signaling pathways underlie this phenomenon, offering valuable insights for development of melanoma therapies.