雄黄微生物提取液对K562／ADM细胞P-糖蛋白和多药耐药基因表达的影响

目的：研究雄黄微生物提取液（RBS）诱导K562／ADM细胞凋亡的作用，并探讨其对P-糖蛋白（P-gP）和多药耐药基因（MDRI）表达的影响。方法：采用“微生物浸出”法制备RBS，并以H3AsO3作为阳性对照，AnnexinV—PI双染法检测细胞凋亡；流式细胞仪检测细胞P—gP表达率；逆转录聚合酶链技术（RT-PCR）检测细胞MDRImRNA表达水平。结果：RBS可诱导多药耐药白血病细胞发生典型凋亡，抑制P-gp的表达，下调MDRI mRNA表达水平。在含砷量相同的条件下，RBS诱导效应明显强于H3AsO3。结论：诱导细胞凋亡、下调P-gp／MDRI蛋白的表达，可能是雄黄逆转白血病耐药的重要分子机制。

Effects of realgar bioleaching solution on the expression of MDRI mRNA and P-gp in K562/ADM cells

AIM:To investigate the role of apoptosis in K562/ADM cells induced by realgar bioleaching solution(RBS),and illustrate the possible molecular mechanism.METHODS:RBS was prepared by a new method of bioleaching with bacteria.The cell apoptosis was determined by annexin V/PI double staining.The expressions of MDRI Mrna and P-gp were detected by RT-PCR and flow cytometry,respectively.The parallel experiments with arsenic acid(H_3AsO_3) were conducted for comparison.RESULTS:RBS inhibited K562/ADM cells growth effectively,and the apoptosis rate of the cells by AnnexinV/PI staining was obviously increased,the expressions of MDRI Mrna and P-gp were significantly down-regulated.With the same concentration of As,the inducing effect of RBS was more stronger than H_3AsO_3.CONCLUSION:RBS induces the apoptosis in K562/ADM cells.The down-regulation of MDRI/P-gp expression may be the important molecular mechanisms in reversing the multi-drug resistance leukemia cells.