The Apoptosis Induction Effect Of Brucine On Human Chronic Myeloid Leukemia Cell Line K562 Cells

ObjeetiveTo investigate the apoPtosis induction effects of brucine on human chronic myeloidleukemia cell line K562 cells.Method1 .To Examine The Growth Inhibition ofBrucine on K562 Cells:K562 cells were exPosedto various dosages ofbrucine.MTT assay was used to examine the growth inhibition ofdifferentconcentration brucine(50,100,200,400,800μg/ml)on K562 cells.2 .Cell Morphofogy Observation of Brucine on K562 Cells:The apoPtosis of K562 cellswas detected by Acridine Orange/Ethidinm Bromlde(AO/EB)double staining.Cells treatedwith the brucine observed under the 490nm excitation wavelength offluorescentmlcroscoPe.3 .To Examine the APoPtosis Induction Effect of Brucine on K562 Cells at the CellularLevel:Changes in cell cycle Progression and DNA Ploidy were examlned before and aftertreatment with brucine of different concentration(50,100,200,400,800μg/m 1) brucine.Two-color irnmunofluorescence flow cytometry was also carried out to examine for oPoPtosis ofbrucine-treated cells.4 .To Examlne the APoPtosis Induetion Effect of Brucine on K562 Cells at the MolecularLevel:Cells treated with the brucine of 400μg/m 1 for 72h and control cells were harvested inorder to confirm the apoPtosis induction effects ofbrucine on human chronic myeloid leukemiacell line K562 cells.ResllltS1 .The Growth Inhibition of Brucine on K562 Cells.The resuits showed that brucine couklremarkabty inhibit the K562 cell growth in a concentration-dePedent andtmle-dePendentmarmer at the ranse of 50 to 800ug·mr,,andt址most significant inhibition 15 at 400μg树ror72h and the inhibition rate 15 to 90.0%.The difference of absorbance vatue in A570nrn was notstatisticalty significant between 400 and 800μg/rnl(P>0.05).The IC50 vatue at 24h,48h,72h oftreatment were about 184μg/ml,107μg/ml,83μg/ml.2 .Staining of cells with AO-EB revealed that brucine induced nuclear chromatincondensation.Afte the K562 cells were treated with the brucine of400μg/rnlfor 72h,the mostof the nucleus was stained orange and condensation-like or bead-llke,was showed apoPtoticmorphology.3 .The APoPtosis Induction Effect ofBrucine on K562 Cells at the Cellular Level.The K562cells treated with brucine of different concentration(50,100,200,400,800μg/ml)for 48h,Annexin一V/Pl apoPtosis detection showed brucine coukl induce apoPtosis of K562 cells.Withthe concentration increase ofbrucine,apoPtosis rate increased graduallv.The K562 cells treatedwith brucine for 72h showed morphofogical characteristics ofapoPtotic cells.4 .The APoPtosis Induction Effect of Brucine on K562 Cells at the Molecular Level:TheK562 cells treated with brucine of different concentration400μg/ml for 72h,and there wastypical ladder strap in DNA gel electroPhoresis.ConClllsion1 .Brucine coukl rernarkabty inhibit the K562 cell growth .The resuits showed that brucinecoukl remarkably inhibit the K562 cell growth in a concentration一dePedent andtime-dePendent rnanner.2 .Brucine coukl induce the K562 cell apoPtosis.The apoPtosis ofK562 cells was detectedby Acridine Orange/Ethidium Broimde(AO/EB)double staining,Annexin-V/ Pl double labelingand DNA agarose gel elelectroPhoresis method,and showed that brucine could induce the K562cell apoPtosis.3 .It will Provide experimental basis for studying molecular mechanism of brucineanti~tumor effect in future.This research Primarity showed that the brucine coukl inhibit theK562 cell growth,and the inhibition may be caused by inducing cell apoPtosis.It will ProvideexPerimental basis for studying molecular mechanism ofbrucine anti-tumor effect in future.