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Characterization of Equilibrative Nucleoside Transport of the Pancreatic Cancer Cell Line: Panc-1.
Appak Basköy, Sila; Khunkhuna, Amardeep; Scuric, Bianca; Naydenova, Zlatina; Coe, Imogen R.
Afiliación
  • Appak Basköy S; Toronto Metropolitan University Faculty of Science, Department of Chemistry and Biology, Toronto, Ontario, Canada.
  • Khunkhuna A; Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Ontario, Canada.
  • Scuric B; University College London, Faculty of Pharmacy, London, United Kingdom.
  • Naydenova Z; Toronto Metropolitan University Faculty of Science, Department of Chemistry and Biology, Toronto, Ontario, Canada.
  • Coe IR; Toronto Metropolitan University Faculty of Science, Department of Chemistry and Biology, Toronto, Ontario, Canada.
Turk J Pharm Sci ; 21(3): 167-173, 2024 Jul 12.
Article en En | MEDLINE | ID: mdl-38994796
ABSTRACT

Objectives:

Gemcitabine, a first-line chemotherapeutic nucleoside analog drug (NAD) for pancreatic cancer, faces limitations due to drug resistance. Characterizing pancreatic cancer cells' transport characteristics may help identify the mechanisms behind drug resistance, and develop more effective therapeutic strategies. Therefore, in this study, we aimed to determine the nucleoside transport properties of Panc-1 cells, one of the commonly used pancreatic adenocarcinoma cell lines. Materials and

Methods:

To assess the presence of equilibrative nucleoside transporter-1 (ENT-1) 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. In addition, we determined the half-maximal inhibitory concentration (IC50) of gemcitabine. Statistical analyses were performed using GraphPad Prism version 8.0 for Windows.

Results:

The 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 s, 62.5 ± 4.8 pmol/mg protein/10 s, and 101.3 ± 2.5 pmol/mg protein/10 s, respectively. Furthermore, the presence of ENT-1 protein was confirmed using NBTI binding assays (Bmax 1.52 ± 0.1 pmol/mg protein; equilibrium dissociation constant 0.42 ± 0.1 nM). Immunofluorescence assays and western blot analysis also revealed ENT-1 in Panc-1 cells. The determined IC50 of gemcitabine in Panc-1 cells was 2 µM, indicating moderate sensitivity.

Conclusion:

These results suggest that Panc-1 is a suitable preclinical cellular model for studying NAD transport properties and potential therapies in pancreatic cancer and pharmaceutical research.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Turk J Pharm Sci Año: 2024 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Turquía
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Turk J Pharm Sci Año: 2024 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Turquía