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

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

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MDRI mRNA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MDRImRNA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。

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 （H3 As03 ） 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 H3 AsO3. 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.