Impact of Di(2-ethylhexyl)phthalate on migration rate of human promyelocytic leukemia cells (HL-60)

Abstract

INTRODUCTION: Softeners like phthalate esters are ubiquitous in the environment and have been detected in transfusion bags, though there is only a limited amount of studies on the effect of phthalates on blood cells. This study seeks to determine effects on cell migration of human promyelocytic leukemia cells (HL-60) incubated with di(2-ethylhexyl)phthalate (DEHP) at concentrations found in blood bags. MATERIAL AND METHODS: HL-60 cells were incubated with DEHP concentrations ranging from 0.1 μg/ml to 1000 μg/ml, diluted in DMSO, over 24 h, 48 h, and 72 h. Migration rate was analyzed along an SDF-1α gradient using Transwell migration inserts. RESULTS: Of the applied concentrations 100 μg/ml, 250 μg/ml, 500 μg/ml, and 1000 μg/ml showed a significant decrease in migration rates relative to DMSO control at all measuring points (p < 0.05), with relative migration rates between 37.87 % for 100 μg/ml and 25.34 % for 1000 μg/ml relative to DMSO after 24 h of stimulation and 19.73 % for 100 μg/ml and 14.69 % for 1000 μg/ml respectively after 72 h of incubation. CONCLUSION: Our results indicate HL-60 to be a suitable in vitro model for examining effects of DEHP on the migration of blood and nucleated cells at concentrations found in blood bags.