论文部分内容阅读
目的 研究粉防己碱抗乳腺癌细胞株多药耐药的作用及可能的抗氧化机制.方法 利用亲代BCap37细胞筛选建立的Bads 200乳腺癌耐药细胞为模型,粉防己碱联合紫杉醇处理Bads 200乳腺癌耐药细胞系24、48、72 h,并设正常培养的对照组以及单纯紫杉醇处理的药物治疗组(单纯药物组).采用MTT法检测各组细胞的增殖活力;流式细胞仪分析各组细胞周期处于各时相的比例;采用反转录酶聚合酶链锁反应(RT-PCR)法检测细胞内皮型一氧化氮合酶(eNOS)及诱导性一氧化氮合酶(iNOS)的表达水平.结果 粉防己碱联合紫杉醇对Bads 200乳腺癌耐药细胞的增殖具有明显的抑制作用,且具有时间相关性.与对照组相比,粉防己碱联合紫杉醇作用24、48、72 h后,Bads 200乳腺癌耐药细胞的凋亡率逐渐增加(P<0.01);S期和G2/M期细胞比例逐渐下降,G0/G1期细胞比例逐渐升高(P<0.01);与对照组相比,粉防己碱组细胞iNOS含量下降,而NO、eNOS含量升高.RT-PCR结果与细胞分光光度计法测定结果相一致.结论 粉防己碱可以抑制Bads 200乳腺癌耐药细胞的增殖,其作用机制可能与抗氧化作用、增加eNOS表达及抑制iNOS表达有关.“,”Objective To research the antagonistic effect of tetrandrine on multidrug resistance of breast cancer cells and the probable antioxidant mechanism.Methods The parental BCap37 cell screening was used to establish Bads 200 breast cancer resistant cell model,then the cells were treated by tetrandrine combined with paclitaxel for 24,48,and 72 hours,respectively.Bads 200 breast cancer resistant cells cultured by normal method and simple paclitaxel were selected as control group and simple drug group,respectively.MTT assay was used to detect proliferation activity of cells in the three groups.Flow-cytometry was used to analyze the proportions of cells during different phases in the three groups.RT-PCR was used to detect the expression levels of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS).Results The inhibiting effect of tetrandrine combined with paclitaxel on proliferation of Bads 200 breast cancer resistant cells was significant,showing a time-dependent manner.Compared with control group,the apoptotic rate of Bads 200 breast cancer resistant cells increased gradually after treated by tetrandrine combined with paclitaxel for 24,48,and 72 hours,respectively (P<0.01).The proportions of Bads 200 breast cancer resistant cells at S stage and G2/M stage decreased gradually,and the proportion of Bads 200 breast cancer resistant cells at G0/G1 stage increased gradually (P< 0.01).Compared with control group,the expression level of iNOS in tetrandrine group decreased,while the expression levels of NO and eNOS increased.The results of RT-PCR were coincidence with the results of cell spectrophotometer method.Conclusion Tetrandrine can significantly inhibit proliferation of Bads 200 breast cancer resistant cells,the mechanism is correlated with antioxidant effect,up-regualtion of eNOS expression,and down-regulation of iNOS expression.