Investigating the effect of differentiation therapy using extracts from ‘Holothuria poli’ (sea cucumber) on the haemopoietic malignancy

Abstract

Leukaemia is a cancer of the blood and is known to affect both males and females all around the world, including Malta. The main research question of this investigation is to study and investigate the potential of an extract from holothurians (sea cucumbers) to be used as a differentiating agent in the treatment of leukaemia. Leukaemia is a cancer which involves a block to the normal differentiation of a whiteblood cell and cause it to proliferate at a very fast rate in immature blast cells. The aim of differentiation is to remove the block allowing the cell to continue its normal maturation and death. A number of H. poli were allowed to eviscerate their intestines and were then allowed to regenerate them in a suitable condition which did not harm the animals. Coelomic fluid was obtained from organisms using a specific bleeding method at various stages of regeneration. MTT and NBT assays were carried out and it was noted whether there were signs of reduced growth with an increase in oxidative burst which show that there is a possibility of differentiation taking place. It was concluded that at day 9 from the evisceration, there was a clear indication that this was happening. It was also confirmed statistically. The fluid was then separated into two fractions; that of < 5 kDa and > 5 kDa and it is has been shown that the effect is a combination of both fractions. Flow cytometry using antibodies has confirmed a change in the surface marker expression of the HL60cell line cells. Marine organisms are known to be a reservoir of biochemicals which are still being discovered and isolated and this investigation will aid research in the treatment of cancer. Hence the results of this dissertation are expected to have an impact on the study and extraction of such differentiation compounds.