Cardamonin Induces Caspase-dependent Apoptosis In Human Multiple Myeloma Cells In Bone Marrow Microenvironment

PART1Cardamonin induces apoptosis in human multiple myeloma cells via Caspase-depent pathway in vitroObjective:To determine the effect of Cardamonin on the viability, proliferation and apoptosis in human multiple myeloma (MM) cells and the involving mechanisms.Methods:CCK-8assay was used to assess the effect of Cardamonin on viability and proliferation of MM cells. Flow cytometry was performed to evaluate the effect of Cardamonin on apoptosis and the change of mitochondrial membrane potential of MM cells. Western blot was applied to survey the effect of Cardamonin on activation of Caspase and PARP whilst on the regulation of pro-apoptosis and pro-survial Bcl-2expression in MM cells.Results:Cardamonin showed inhitited viability and proliferation of MM cells, which was revealed by CCK-8assay in a dose-and time-denpendent manner. Flow cytometry exhibited that Cardamonin induced apoptosis and decline of mitochondrial membrane potential of MM cells. Western blot relealed that Cardamonin induced activiation of Caspase and PARP of MM cells,whilst down-regulation of pro-survival Bcl-2family protein expression and up-regulation of pro-apoptosis Bcl-2family protein expression. A pan Caspase inhibitor, Z-VAD-FMK, showed suppressive effect on inhibitory activity of Cardamonin angaist MM cells, induction of apoptosis and activation of Caspase and PARP in MM cells.Conclusion:Cardamonin supresses viability and proliferation and induced apoptosis of MM cells via Caspase-dependent pathway. PART2Cardamonin induces apoptosis in human multiple myeloma cells in bone marrow microenvironment in vitroObjective:To determine the effect of Cardamonin on the viability, proliferation and apoptosis in human multiple myeloma (MM) cells in bone marrow microenvironment in vitro and the involving mechanisms.Methods:Co-culture of IL-6, VEGF or bone marrow stromal cells (BMSCs) and MM cells to simulate bone marrow miroenvironment. CCK-8assay was used to assess the effect of Cardamonin on viability and proliferation of MM cells. Flow cytometry was performed to evaluate the effect of Cardamonin on apoptosis and the change of mitochondrial membrane potential of MM cells. Western blot was applied to survey the effect of Cardamonin on activation of Caspase and PARP.Results:With or without co-culture of MM cells and IL-6, VEGF or BMSCs, Cardamonin showed similar inhititory activity against viability and proliferation of MM cells which was evaluated by CCK-8assay, notable apoptosis and decline of mitochondrial membrane potential releaved by Flow cytometry, and relealed that Cardamonin induced alike activiation of Caspase and PARP by Western blot.Conclusion:Cardamonin supresses viability and proliferation and induced apoptosis of MM cells in bone marrow microenvironment in vitro.