Preparation and interaction of a low-density lipoprotein:daunomycin complex with P388 leukemic cells

Abstract

A low-density lipoprotein (LDL):daunomycin complex was prepared which contains, on the average, 8 nmol of daunomycin/mg of apolipoprotein. LDL was not perturbed to any significant degree by association with daunomycin, and the complex remained stable at 4 degrees. Fluorescence quenching with DNA and potassium iodide, antibody precipitation, and density flotation studies suggested at least two domains for localization of the daunomycin, the surface region and the hydrophobic core of the LDL particle. Interaction of the LDL:daunomycin complex with P388 leukemic cells which were sensitive or resistant to daunomycin occurred rapidly in both sublines, with quantitatively larger amounts of the drug being cell associated than when free drug was used alone. The cells appeared morphologically intact, with no nuclear damage after short-term incubation with LDL:daunomycin. Subfractionation of cell membranes and organelles from the sensitive and resistant cell lines incubated with LDL:daunomycin showed accumulation of drug in the plasma membrane and microsomal-lysosomal-mitochondrial membranes and an insoluble nuclear chromatin material. Both 125l-labeled apoprotein and daunomycin from a 125l-LDL:daunomycin complex showed enrichment in similar membrane:organelle fractions of the P388 cells.