NSC-631570 alkaloid mixture modulates the human equlibrative transporter nucleoside 1 and deoxycytidine kinase gene expressions in pancreatic ductal adenocarcinoma cell cultures

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is treated by palliative chemotherapy for advanced disease. Gemcitabine is the standard drug in both adjuvant and palliative treatment, but yields a marginal impact on disease outcome. Several attempts to improve the efficacy of gemcitabine by addition of a second cytotoxic or targeted agent have not shown a significant survival advantage. NSC-631570, showed greater median survival in combination with gemcitabine compared to gemcitabine alone in the palliative treatment of unresectable PDAC. However, the authors did not study the interactions between alkaloids and molecular determinant expressions involved in the metabolism of gemcitabine. Therefore, the aim of present study was to evaluate the modulation of the expression of two pivotal genes (hENT1 and dCK) involved in gemcitabine activity. In vitro studies on cell lines and primary cell Cultures from PDAC patients were treated with NSC-631570 at IC50 concentration for 48h. the quantitation of gene expression was performed using the standard curve method and the ΔΔCT calculation. Alkaloids mixture positively modulates the expression of hENT1 mRNA in all PDAC cell cultures. The 2 (-ΔΔCt) analyses revealed a mean increase of 2.8 fold with respect to untreated control cells. In two cell lines cells alkaloids positively affects mRNA expression of dCK gene as well. Based on the previous clinical data the Alkaloids-gemcitabine combination appears a promising regimen and the results of the present study provide the experimental basis for the further clinical testing of the NSC-631570-gemcitabine schedule in PDAC patients.