Inhibition Efefct And Mechanism Of20(S)-ginsenoside Rg3in Vitro On Multiple Myeloma Cell Line U266

Objective:To study the natural compounds20(S)-ginsenoside Rg3in vitro on multiple myelomacell line U266proliferation inhibition, analysis of effects,of20(S)-ginsenoside Rg3onmyeloma cells secreting VEGF, IL-6; illustrate mechanism of20(S)-ginsenoside Rg3inhibitsmyeloma cell proliferation, provide theoretical and experimental basis about20(S)-ginsenoside Rg3as a novel therapy drugs for multiple myeloma.Material and methods:To detecte the inhibition rate through MTT method after the U266cell was treated withdifferent doses of20(S)-ginsenoside Rg3in24h,48h and72h; To detect the cell circles andapoptosis after treated with20(S)-ginsenoside Rg3for24h at different concentrations flowCytometry(FCM) and the effect of apoptosis was observed by Wright-Giemsa staining in oilmicroscope; the application of Western blot to detect the Caspase-3, Caspase-8, caspase-9protein expression influence after20(S)-ginsenoside Rg3on multiple myeloma cell lineU266for24h; the concentrations of interleukin-6and vascular endothelial growth factor ofthe supernant were measured by ELISA.Results:The results showed that within a certain range of concentration, IC50of20(S)-ginse-noside Rg3to U266at24hours,48hours and72hours were71.07±2.63μmol/L,44.06±3.98μmol/L and25.93±2.22μmol/L,respectively.Its inhibiting effect on prolife rationof U266showed both time-and dose-dependent manners(P <0.05); Flow cytometry withAnnexin-V/PI double staining revealed that20(S)-ginsenoside Rg3may induce multiplemyeloma U266cells apoptosis and in a concentration-dependent manner(P <0.05);Drugtreatment group appeared chromatin condensation in the cells, nuclear edge set and typicalapoptotic bodies, apoptotic appearance under oil microscope Wright-Giemsa staining; Flowcytometry with PI staining showed that the ratio of cells in G0/G1phase increased(P <0.05),while it in S phase decreased(P <0.05),and ratio of cells at G2/M phase did not presenta significant change(P>0.05); U266cells were treated after24hours20(S)-ginsenosideRg3,Western blot detection that with drug concentration increased, procaspase-3,-8,-9,a milddecline, cleaved-caspase-3,-8,-9significantly increased expression(P <0.05); Used different concentrations of the drug treatment of cells by24h, VEGF and IL-6volume showed adose-dependent reduction in cell culture supernatant (P <0.05).Conclusion:In a certain range of concentration,20(S)-ginsenoside Rg3inhibits proliferation ofU266cells and showed both time-and dose-dependent manners;20(S)-ginsenoside Rg3caninduce apoptosis of U266cells through the excitation/exogenous apoptotic pathway andshowed both time-and dose-dependent manners;20(S)-ginsenoside Rg3can also affectU266cell line G1/S phase, cells arrest in the G1phase, and the G2/M had no significanteffect, which may be one of the mechanisms of tumor suppression;20(S)-ginsenoside Rg3can inhibit the secretion of VEGF and IL-6of U266cells； Makes it a potential therapy formultiple myeloma with new agents.