扁塑藤素对非小细胞肺癌增殖及线粒体膜电位的调节作用

Regulation of Pristimerin on Proliferation and Mitochondrial Membrane Potential of Non-small Cell Lung Cancer

目的分析扁塑藤素对体内外非小细胞肺癌增殖及线粒体膜电位的调节作用,初步探讨扁塑藤素的抑瘤增殖效果及可能机制.方法将适量的扁塑藤素提取物分别作用于非小细胞肺癌细胞系(A549及H226)及动物模型(BALB/c裸鼠),应用MTT方法测定其细胞存活率,应用细胞集落实验检测其长期毒性作用.制备A549非小细胞肺癌裸鼠模型,测定PTM对移植瘤体积及湿重的生长抑制.应用流式细胞术测定PTM共培养后细胞线粒体膜电位.结果与空白对照组比较,A549和H226两组细胞随PTM剂量的增加及共培养时间的延长细胞的平均存活率显著降低.与对照组(PTM:0μmol/L)比较,0.1μmol/L组A549及H226的细胞集落形成均受到抑制,并且0.2μmol/L组的细胞集落形成抑制更为明显.PTM腹腔内注射治疗第7天,PTM 2 mg/kg治疗组的移植瘤(A549)体积抑制作用优于PTM 1 mg/kg治疗组(P<0.05).湿重对比分析结果显示:PTM 2mg/kg治疗组移植瘤湿重低于空白组及PTM 1 mg/kg治疗组(P<0.05).流式细胞术测定结果显示:PTM(8μmol/L)对各组细胞(A549和H226)作用时间持续3 h即可导致细胞线粒体膜电位下降,持续6 h时线粒体膜电位下降更为明显,提示PTM作用时长与细胞(A549和H226)线粒体膜电位呈负性相关.结论 PTM对体内外非小细胞肺癌(A549和H226)的细胞抑制作用存在剂量和时间依赖性,并初步证明PTM具有诱导细胞线粒体去极化的作用.

Objective To analyze the regulation effects of pristimerin on the proliferation and mitochondrial membrane potential of non-small cell lung cancer in vivo and in vitro,and to preliminarily explore the suppressive effects of pristimerin on tumor proliferation and its possible mechanism.Method Appropriate amount of pristimerin extract was applied to non-small cell lung cancer cell lines(A549 and H226)and animal models(BALB/c nude mice),respectively.The cell viability was determined by MTT method.The long-term toxicity was detected by cell colony formation.A nude mouse model of A549 non-small cell lung cancer(NSCLC)was prepared to determine the growth inhibition effects of PTM on transplanted tumor volume and wet weight.The cell mitochondrial membrane potential was determined by flow cytometry after PTM co-culture.Results Compared with the blank control group,the average survival rate of A549 and H226 cells decreased significantly with the increase of PTM dose and the prolongation of co-culture time.Compared with the control group(PTM:0μmol/L),the cell colony formation of A549 and H226 in the 0.1μmol/L group was inhibited,and the cell colony formation inhibition in the 0.2μmol/L group was more obvious.On the 7th day of intraperitoneal injection of PTM,the volume suppression effect on the transplanted tumor(A549)in the PTM 2 mg/kg treatment group was better than that in the PTM 1 mg/kg treatment group(P<0.05).The comparative analysis of wet weight showed that the wet weight of transplanted tumor in the PTM 2 mg/kg treatment group was lower than that in the blank group and the PTM 1 mg/kg treatment group(P<0.05).The results of flow cytometry showed that the effect time of PTM(8μm ol/L)on cells(A549 and H226)in each group lasted for 3 h,which resulted in the decline of cellular mitochondrial potential,and the decline of mitochondrial potential was more obvious when it lasted for 6 h,sug-gesting that the duration of PTM action was negatively correlated with the mitochondrial membrane potential of cells(A549 and H226).Conclusion There was a dose-and time-dependent correlation between PTM and cell inhibition of non-small cell lung cancer(A549 and H226)in vivo and in vitro,and it was preliminarily proved that PTM has the effect of inducing cell mitochondrial depolarization.