ANTI TUMORAL AND DIFFERENTIATION EFFECTS OF ALKALOIDS OF HARMINE AND HARMALINE ON LEUKAEMIC CELLS TREATED WITH ATRA AND G-CSF

In acute leukaemia, the maturation of the malignant cells is arrested and the cells merely proliferate. In the recent years, beside chemotherapy combination of differential factors cytokins and cytotoxic agents have been used in treatment of acute leukaemia particularly acute promyelocytic leukaemia. The present study was an evaluation of proliferation, cytotoxicity and differentiation of Harmine and Harmaline as alkaloid derivates of Peganum Harmala on HL60 cells individually or in combination with ATRA and G-CSF. The data showed that these agents caused cessation of proliferation in dose and time dependent manner. Optimal concentration of anti proliferative effect was 0.4, 0.8 and 1.6μg/mL for Harmine and 6-10 μg/mL for Harmaline. However, both agents in concentration of over 6.4 and 10 μg/mL for Harmine and Harmaline were cytotoxic respectively. Combination of ATRA(10-7 M) and G-CSF(100 ng/mL) with each optimal concentration of agents extensively reduced proliferation compared with ATRA and G-CSF. Cells, induced with optimal concentration of these agents, showed some morphological changes and NBT positivity however, it was in lesser extent compared to ARTA and G-CSF. Most overt sign of myeloid differentiation was observed using 10 μg/mL Harmaline. Fluorescent analysis showed an increase in CD11b and CD14 using Harmaline. Combination of ATRA and G-CSF with each agent caused differentiation similar to ARTA and G-CSF. These preliminary data showed that either Harmine or Harmaline in optimal and non toxic dose caused cessation in proliferation and some degree of differentiation using Harmaline. These results have opened a new window in the leukaemic in vitro investigation.