Characterization of equilibrative nucleoside transport of the pancreatic cancer cell line: Panc-1

INTRODUCTION: Gemcitabine is a nucleoside analog drug used as a first-line chemotherapeutic agent against pancreatic cancer. The characterization of cell lines is crucial for understanding drug resistance development in pancreatic cancer. In this study, we aimed to determine the nucleoside transport properties of Panc-1 cells, one of the commonly used pancreatic adenocarcinoma cell lines.
METHODS: To assess the presence of Equilibrative nucleoside transporter 1 (ENT1) in Panc-1 cells, we performed immunofluorescence staining, western blot analysis, and S-(4-nitrobenzyl)-6-thioinosine (NBTI) binding assays. We also conducted standard uptake assays to measure the sodium-independent uptake of [3H]-labeled chloroadenosine, hypoxanthine, and uridine. Additionally, we determined the half-maximal inhibitory concentration (IC50) of gemcitabine. Statistical analyses were performed using GraphPad Prism version 8.0 for Windows.
RESULTS: Sodium-independent uptake of [3H]-labeled chloroadenosine, hypoxanthine, and uridine was measured using standard uptake assays, and the transport rates were determined as 111.1 ± 3.4 pmol/mg protein/10 secs, 62.5 ± 4.8 pmol/mg protein/10 secs, and 101.3 ± 2.5 pmol/mg protein/10 secs, respectively. Furthermore, the presence of ENT1 protein was confirmed using S-(4-nitrobenzyl)-6-thioinosine (NBTI) binding assays (Bmax 1.52 ± 0.1 pmol/mg protein; Kd 0.42 ± 0.1 nM). Immunofluorescence assays and western blot analysis also showed the presence of ENT-1 in Panc-1 cells. The determined IC50 of gemcitabine in Panc-1 cells was 2 μM, indicating moderate sensitivity.
DISCUSSION AND CONCLUSION: These results suggest that Panc-1 is a suitable preclinical cellular model for studying nucleoside analog drug transport properties and potential therapies in pancreatic cancer and pharmaceutical research.