Abstract
Objective: Three-dimensional (3D) culture systems offer a more physiologically relevant environment than conventional two-dimensional (2D) cultures, particularly for studying tumor biology. Here, we systematically compared two widely used 3D platforms-poly(2-hydroxyethyl methacrylate) (Poly-HEMA, PH)-coating and ultra-low attachment (ULA) plates-to evaluate their impact on pancreatic cancer (PCa) cell behavior. We assessed spheroid morphology, chemotherapeutic response, invasion potential, and adhesion molecule expression in two PCa cell lines (PANC-1 and SU.86.86). Results: Spheroid morphology differed markedly between PH and ULA platforms, with ULA generally promoting larger and more cohesive spheroids. Gemcitabine resistance was the highest in SU.86.86 spheroids on ULA plates. Matrigel invasion assays revealed enhanced single-cell migration in SU.86.86 spheroids grown on PH, whereas ULA spheroids exhibited broader matrix degradation and collective invasion. Moreover, gene and protein expression levels of key adhesion molecules, including E-Cadherin, N-Cadherin, and integrins, varied between platforms in both cells. These findings demonstrate that 3D culture systems distinctly influence PCa cell characteristics and highlight the necessity of selecting appropriate culture environment for studying tumor biology and drug response. Further mechanistic studies are warranted to uncover how different 3D environments shape tumor cell behavior and therapy resistance.