Studies On Anti-cancer Effect Of Annonaceous Acetogenin On Melanoma And Its Mechanism

Annonaceous acetogenins is the characteristic ingredients of the Annonaceae plant and have a broad and strong anti-tumor effect both in vitro and in vivo. This article does further systematic study to investigate the activity of Annonaceous acetogenin derivative components against melanomain in vitro and its mechanism, so as to provide theoretical basis for the development and utilization of plant resources.1、The inhibiting proliferation of the crude extract AS9and monomer compounds Desacetyluvaricin, Squamocin, Bullatacin. In vitro screening adopt SRB method to detect the optical density value, IC50value was calculated to determine the sensitive strains. The results show that:the crude extract AS9and monomer compounds Desacetyluvaricin, Squamocin, Bullatacin have different cytotoxic effect on different melanoma cells. The antitumor effect of Desacetyluvaricin was the best for human melanoma cell A375, the IC50value was0.71μg/mL; Squamocin has the best antitumor effect on human melanoma cell A875,which IC50value is0.06μg/mL; The AS9and Bullatacin showed good concentration dependent manner on B16cells, the IC50values of which were5.93μg/mL、2.07μg/mL, respectively. AS9, Desacetyluvaricin, Bullatacin, Squamocin can significantly inhibit the growth of A875, A375, B16melanoma cells.2、The inhibitory mechanism study of the crude extract AS9on melanoma cell A875. H33342/PI fluorescent stain was assayed to detect the morphological changes of nucleus. Fluorescent microscopic,with the increase of concentration, A875nucleus has undergone chromatin condensation, nuclear shrinkage and marginalization. Dense, granular or block fluorescence appeared within a cell’s nucleus. Many nuclei have become fragments karyopyknosis. The relationship between LDH level and the rate of cell viability was investigated by determining the activity of lactic dehydrogenase (LDH). The results showed that, with the concentration of AS9increase, the release rate of lactate dehydrogenase increases. Content with dead cells in the medium LDH was proportional relationship. The cell cycle and apoptosis rate of A875melanoma cells incubated with AS9was analysed by Annexin V/PI double and single staining flow cytometrty methods. The results show that AS9mainly induced apoptosis of A875cells and arrested cells in G1Phase in a dose-dependent manner.3、The study on the cell cycle and apoptosis of monomer compound Desacetyluvaricin acted on A375cell. Flow cytometry analyses shows the Desacetyluvaricin has significant effect on inhibiting the cell proliferation of melanoma cell, and hold back cell course from S to G2, and induced A375apoptosis.