Research On Apoptosis Of K562 Tumor Cells Induced By Several Venom Components Of Bungarus Multicinctus

Aim: TO make sure whether Buoprus multicinctus crude venom induces theaPoptosis of K562 tumor cells of CML or not, to find out the components inducingaPoptosis of K562 cells from the crude venom, and to make certain of the time rangeand concentfation in which the crude venom can kill K562 cells largely Theconclusions maybe provide a new tool for the research or therapy of leukaemia andtheory fOr the exploitation of snake venom.Method8: The paPef, fOr the first time, studied the apoptosis or necrosis of K562cclls induced by bungarus multicinclus crude venom and several components frOmcrude venom by cation exchange chromatography' through the quatitative analysis offlow cytometry as well as the qualitative analysis, such as inversed opticalmicroscopy, MTT method and fluorescent microscopy, and so on.Re8ult8: In the MTT assay, the absorbency and livability of the K562 cells is thesame high as the blank control group in 24 or 48 or 72 hours after administration ofthe crude venom of below 8 X l0'nglml, but a rapidly decrease occurred afteradministration of 8 X l0'ng/ml toxin or 5 P g/ml Cycloheximide (CHX). theabsorbency and livability of the K562 ce1ls is the saxne low as the blank control groupin 96 or l20 hours after administration of the crude venom of below 8 X l0'ng/mL,but a significantly increase was observcd after administration of 8 X l0'nglml toxin,and following by the decreasing after administration of 8 X l0'ng/ml crude venom.APOptotic bodies were observed among the K562 tumor cells in fluorescentmicroscopy aftcr administmtion of 5 v glml C14X or the sixth peak of about 8Xl0'ng/ml. The same results were detected by the flow cytometer' In the figures, asub-Gl pcilk appearcd bcfOrc tl1c GI cells group after adn1inistration of CHX or tl1e3sixth peak solution.Conclusions: (l) The bungarus n1ulIicinclus crude venom can kill K562 tumorcells of human CML or restrain the growth of them in the time- and dose-dependentmanners, and the effect is significant in 24 hours when administrating 8 X l0'ng/mlcrude toxin. (2) It is difficult to make sure whether apoptosis takes place or not. (3)Only the sixth peak solution can induce apoptosis among the several componentsseparated from the crude venom. (4) Q -BTX can't induce apoptosis of K562 cells. (5)the aPoptosis-inducing factors, of the toxins in the sixth peak whose content is 2.2%of the crude venom, may be kappa-bungarotoxin or an unknown protein whosemolecule wcight is about 20 kDa.