Abstract 5651: A novel chemopreventive regimen synergistically affects cell viability and apoptosis in Panc-1 and MIA PaCa-2 pancreatic cancer cell lines

Abstract

Abstract Cancer of the pancreas is the fourth leading cause of cancer deaths in the United States with approximately 42,270 estimated diagnosed cases in 2009. The prognosis for most patients is grim with high mortality rates. The objective of our study was to study the chemopreventive effects of a combined regimen of the non-steroidal anti-inflammatory (NSAID) drug aspirin with sulforaphane and curcumin when administered in low doses to two pancreatic cancer cell lines, Panc-1 and MIA PaCa-2. Effects of the drug combination were determined by the use of MTS cell proliferation assay and apoptosis analysis. For the MTS assay, the Panc-1 and MIA PaCa-2 cells were cultured as per established protocols. Upon 75 % confluence, 4 × 103 cells for Panc-1 and 2.5 × 103 cells for MIA PaCa-2 were transferred into each well of 96-well plates. The aspirin, sulforaphane and curcumin chemopreventive agents alone or in combination were added to the incubated cells the following day. The cells were incubated with drugs for a period of 72 h. Absorbance was recorded at 490 nm using an ELISA plate reader. For the apoptosis assay, 3 × 105 cells were cultured in 6-well plates for both Panc-1 and MIA PaCa-2 cell-lines. Drugs of interest were added the next day and the plates were incubated for 72 h, same as the MTS Assay. FITC-Annexin V and propidium iodide were then added as per established protocols and the cells were analyzed using a flow cytometer, measuring the fluorescence emission at 530 nm (FL1) and >575 nm (FL3). Panc-1 cells treated with aspirin (1 mM) combined with sulforaphane (5 μM) and curcumin (10 μM) showed a significant decrease in cell viability of > 75% using the MTS assay. Flow cytometry analysis showed that individual concentrations (aspirin 1 mM, sulforaphane 5 μM and curcumin 10 μM) of each drug demonstrated low apoptotic effects (18-21% effect) but when combined, approximately 63% apoptosis was observed. Thus, combined use of lower concentrations of the chemopreventive regimens demonstrated significant apoptotic effect. With the MIA PaCa-2 cells, treatment with similar concentrations of aspirin (1 mM), sulforaphane (5 μM) and curcumin (10 μM) showed a >60% decrease in cell viability (MTS assay) and 51% induction of apoptotic cells indicating that these chemopreventive regimens in combination were significantly effective against both cell lines. Our results demonstrate, for the first time, low doses of a novel chemopreventive combination of drugs are synergistically effective in decreasing cell proliferation and inducing apoptosis of pancreatic cells. This new data provides compelling evidence of the potential in designing novel regimens for pancreatic cancer chemoprevention. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5651.