Arsenic Trioxide Combined With Bso On K562/adm Cells Induced Apoptosis Research

Objective To investigate the apoptosis-inducing effect and P-gp expression of arsenic trioxide (As₂O₃ ) and buthionine sulfoximine (BSO) on multidrug-resistant K562/ADM cell. To compare the effect of As₂O₃ and the combined group .To discuss the reverse effect and the reverse mechanism of the combined group of clinic dose arsenic and BSO on multidrug resistance. Methods To investigate the effect of clinic dose arsenic group and high dose group combined BSO (100μmol/L) on human leukemia K562/ADM cell. To detect the change of the correlated index.(1) The cell proliferating activity was assessed with MTT assay;(2) The cell apoptosis effect was detected by Annexin-V and propidium iodide ( PI) staining .(3) Intracellular GSH contents were measured using Glutathione Assay Kit.(4) P-gp expression were determined by flow cytometry.(5) Mdr-1mRNA expression were directed by semi-quantitative RT-PCR.Results(1) MTT assay disclosed that followed by the extension of effect time of clinic dose arsenic group , the cell depressive effect was degraded(P﹤0.05). On the other hand ,followed by the extension of effect time of high dose group , the cell depressive effect was advanced.(P﹤0.05).After the GSH contents were degraded by the combination of clinic dose arsenic group and BSO, the K562/ADM cell proliferating activity was obviously inhibited(P﹤0.05). Compared with the high dose group, deviation in 24 hours was not significant(P>0.05).In 48 hours and 72 hours, the cell depressive effect of the combination group (clinic dose arsenic group ) was obviously stronger than high dose arsenic group(P﹤0.01).(2) Annexinâ…¤/PI assay disclosed that followed by the extension of effect time of clinic dose arsenic group , the cell apoptosis effect was degraded(P﹤0.05). On the other hand ,followed by the extension of effect time of high dose group , the cell apoptosis effect was advanced(P﹤0.05).The K562/ADM cell apoptosis effect was obvious(P﹤0.05),after the GSH contents were degraded by the combination of clinic dose arsenic group and BSO. Compared with the high dose group, deviation in 24 hours was not significan(tP>0.05).In 48 hours and 72 hours, the cell apoptosis effect of the combination group (clinic dose arsenic group ) was obviously stronger than high dose arsenic group(P﹤0.01).(3) Spectrophotometric method disclosed that,in 24 hours,the clinic dose arsenic group and...