Centipede On Isolation, Purification And Antitumor Protein Pharmacological Mechanism

ObjectiveCentipede (Scolopendra subspinipes mutilans L. Koch) has good clinical grounding in anti-tumor in China. The present study is aimed to purify the major anti-tumor proteins extracted from centipede,and to study the possible anticancer pharmacological mechanisms of proteins compound CGI extracted from centipede.MethodsThe proteins were purified through precipiting with (NH4)2SO4, followed by gel filtration chromatography of Sephadex G-25, ion chromatography of Q Sepharose fast flow and gel filtration chromatography of Sephadex G-200. And MTT colorimetric method was applied to observe the main anti-tumor components. The possible pharmacological anticancer mechanisms of CGI were studied on the tumor cells of Hela and BGC-823 at two points of view, of which one is inducing apoptosis and the other is cytotoxicity. For the first, three staining skills including Giemsa, AO/EB and AO fluorescent dyes,agarose gel electrophoresis technique of DNA were adopted. For the second, MTT colorimetric method, the leakage percentage of LDH and the quantity of protein in the tumor cells were measured.ResultsOur research has purified the anti-tumor proteins extracted from centipede in same way for the first time. The results of SDS-PAGE showed that the half-purified proteins we got still have 8 components at less. And its suppressive effect on the proliferation of Hela was nearly nine times more than the crude proteins of centipede.The results of MTT showed that the proteins compound CGI have remarkable suppressive effect on the proliferation of Hela and BGC-823 in vitro. And there was better dose-dependent relationship. Besides, CGI could remarkably increase the leakage percentage of LDH and decrease the quantity of protein in those two tumor cells. However, there was no apoptosis grain was observed in the results of stains and no DNA Ladder in the agarose gel electrophoresis.ConclusionThe protein compound we got is worth further purification.The protein compound CGI extracted from centipede inhibited markedly the proliferation of Hela and BGC-823. And its possible anti-tumor pharmacological mechanism is likely killing the tumor cells through interfering structure and function of cell membrane,but not inducing apoptosis.