Objective To investigate the effect of schizandrin A on drug resistance of glioma stem/ progenitor cells (GSPCs) and its mechanism.Methods Isolate and culture SHG-44s cells from human glioma cell line SHG-44.The SHG-44s cells were treated with different concentrations of schizandrin A (0,12.50,25.00 and 50.00 μmol/L) and vincristine (400,800 and 1200 nmol/L).The cell proliferative activity was measured by cell counting kit-8 (CCK-8) assay.Rhodamine 123 staining was used to detect the drug delivery ability of SHG-44s cells.The transcription and translation ability of ATP binding cassette subfamily B member 1 (ABCB1) gene of SHG-44s cells was detected by real-time polymerase chain reaction (PCR) and Western blotting.Results The proliferative activity of SHG-44s cells was inhibited when the concentration of schizandrin A was 50 μmol/L (P =0.001,0.001,0.039),so this concentration was removed in the follow-up study.No matter the concentration of vincristine was 400,800 or 1200 nmol/L,combining with schizandrin A could inhibit the proliferative activity of SHG-44s cells (vincristine 400 nmol/L:P =0.007,0.001;vincristine 800 nmol/L:P =0.001,0.000;vincristine 1200 nmol/L:P =0.000,0.000).Inverted fluorescence microscopy findings showed SHG-44s cells in the group of schizandrin A 0 μ mol/L rarely revealed green fluorescence,while SHG-44s cells in the groups of schizandrin A 12.50 and 25.00 μmol/L presented obvious green fluorescence.Flow cytometry showed that with the increasing of schizandrin A concentration,the percentage of positive cells by Rhodamine 123 staining was 10.40％,39.20％ and 45.20％,respectively.Real-time PCR showed that ABCB1 gene expression levels of SHG-44s cells in schizandrin A 12.50 μmol/L group and 25.00 μ mol/L group were significantly decreased comparing with schizandrin A 0 μmol/L group (P =0.027,0.006),especially in schizandrin A 25.00 μmol/L group (P =0.034).Western blotting showed that the expression level of P-glycoprotein (P-gp) in SHG-44s cells was gradually decreased with the increasing of schizandrin A concentration.Conclusions Schizandrin A can inhibit the drug delivery ability of P-gp coded by ABCB1 gene existing in the surface of GSPCs.It can further reverse the drug resistance of GSPCs by reducing the transcription and translation of ABCB1 gene.%目的 探讨五味子甲素对胶质瘤干/祖细胞耐药性的影响及作用机制.方法 自人胶质瘤细胞系SHG-44中分离培养胶质瘤干/祖细胞SHG-44s,予五味子甲素0、12.50、25.00和50.00 μmol/L联合长春新碱400、800和1200 nmol/L,细胞活性检测试剂盒CCK-8细胞毒性实验检测SHG-44s细胞增殖活性,罗丹明123染色检测SHG-44s细胞泵出药物能力,实时聚合酶链反应(PCR)和Western blotting法检测SHG-44s细胞ATP结合盒转运子B1(ABCB1)基因转录和翻译能力.结果 五味子甲素50 μmol/L即可抑制SHG-44s细胞增殖活性(P=0.001,0.001,0.039),剔除这一浓度后无论长春新碱浓度为400、800或1200 nmol/L,联合应用五味子甲素均可抑制SHG-44s细胞增殖活性(长春新碱400 nmol/L组:P=0.007,0.001;长春新碱800 nmol/L组:P=0.001,0.000;长春新碱1200 nmol/L组:P=0.000,0.000).倒置荧光显微镜观察,五味子甲素12.50 μmol/L组和25.00 μmol/L组SHG-44s细胞可见明显绿色荧光.流式细胞术显示,随着五味子甲素浓度的增加,SHG-44s细胞罗丹明123染色阳性细胞比例分别为10.40％、39.20％和45.20％.实时PCR法显示,五味子甲素12.50μmol/L组和25.00μmol/L组SHG-44s细胞ABCB1基因表达水平较0μmol/L组降低(P=0.027,0.006),尤以25.00μmol/L组显著(P=0.034).Western blotting法显示,随着五味子甲素浓度的增加,SHG-44s细胞P-糖蛋白表达水平下降.结论 五味子甲素通过抑制胶质瘤干/祖细胞表面已存在的ABCB1基因编码的P-糖蛋白泵出药物能力并降低ABCB1基因转录和翻译能力,逆转胶质瘤干/祖细胞耐药性.
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