Effects of Polycyclic Aromatic Hydrocarbon-Containing Oil Mixtures on Generation of Reactive Oxygen Species and Cell Viability in MCF-7 Breast Cancer Cells

Abstract

Clarified slurry oil (CSO), and two crude oil samples, Belridge heavy crude oil (BHCO) and Lost Hills light crude oil (LHLCO), were examined for their ability to generate reactive oxygen species (ROS) in MCF-7 cells. Intracellular ROS and cell viability were determined in a flow cytometer using dihydroxyrhodamine 123 and propidium iodide, respectively. In experiments with short-term exposure, single-cell suspensions were loaded with the fluorescent probes and then treated with the oil samples (1 or 10 ppm). Measurements were made at 5, 15, 30, 60, and 90 min after addition of oil samples. In experiments with longer term exposure, preconfluent cell cultures were treated with oil samples for 6, 12, or 24 h prior to preparing single-cell suspensions. Both short-term and longer term treatment with oil samples resulted in elevated generation of reactive oxygen species (ROS). Cell cultures also were treated with benzo[a]pyrene, a polycyclic aromatic hydrocarbon detected in all three oil samples. Treatment with benzo[a]pyrene produced a significant increase in levels of ROS. The present findings suggest that oil samples with higher concentrations of polycyclic aromatic hydrocarbons may exert adverse effects on human mammary epithelial tissue through induction of oxidative stress.

This study was supported by NIEHS award RO1 ES049795-01A1 (to KA), NIH Fogarty International Center 5D43TW00636 (to DOC), and a TUBITAK-NATO fellowship (to BY).