The effect of membrane active agents on human leukaemia cells

Abstract

This Thesis investigates the effect of membrane-active agents, such as synthetic ether lipids (SEL), local anaesthetics and polyunsaturated fatty acids (PUFAs) on human leukaemia cells. The two cell lines used were human acute myeloblastic leukaemia (HL60) cells and human myelogenous leukaemia (K562) cells. SEL, local anaesthetics and PUFAs were found to be cytotoxic to both cell lines at certain concentrations. The SEL ET-18-OCH(_3) was found to be cytotoxic to both cell lines but the HL60 cells were found to be the more sensitive cell line. HL60 cells were found to be so sensitive to the action of the local anaesthetic dibucaine that a subtoxic concentration that killed ≤10% was not determined. However, in K562 cells the combination of a subtoxic dibucaine concentration together with a range of ET-I8-OCH(_3) concentrations increased the cytotoxicity over that of ether lipid alone. PUFAs were shown to incorporate into plasma membrane phospholipids at concentrations as low as 1 μM after an incubation of 48 hours. PUFAs were shown to be cytotoxic, but the addition of vitamin E reduced the cytotoxicity of arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid in HL60 cells, and of docosahexaenoic acid in K562 cells. This implied that lipid peroxidation was involved in PUFA cytotoxicity. This was, however, not confirmed. PUFA in combination with ET-I8-OCH3 resulted in a slight decrease in cytotoxicity. PUFA combined with dibucaine did not alter cytotoxicity. Cells were also treated with a combination of PUFA and 1-β-D- arabinofliranosylcytosine (ara-C), which is an agent known to induce cell differentiation. Onset of differentiation was determined by following haemoglobin accumulation in K562 cells. PUFA on their own were found to promote accumulation of haemoglobin. The greatest accumulation of haemoglobin was observed with K562 cells treated with PUFA and ara-C.