鸦胆子油乳对Hep-2细胞增殖与凋亡诱导作用的研究

Effect of Brucea javanica oil emulsion on proliferation and apoptosis of human laryngeal cancer Hep-2 cells

目的:探讨鸦胆子油乳注射液对喉癌Hep-2细胞的增殖抑制及调亡诱导作用。方法:利用MTT比色法检测5个不同药物浓度(0.1、0.2、0.4、0.6和0.8g/L)对细胞的抑制率;采用倒置显微镜观察细胞的形态变化;流式细胞仪检测细胞凋亡率;实时荧光定量PCR检测凋亡基因Bcl-2和Bax的表达。结果:MTT显示,鸦胆子油乳对Hep-2细胞的生长抑制作用呈剂量和时间依赖性。倒置显微镜下可见细胞凋亡的形态学改变,显示加药组细胞染色体固缩,聚集呈块状、新月状,出现凋亡小体。流式细胞仪检测显示,0.2、0.4和0.6g/L药物作用48h后凋亡率分别为(42.58±4.36)%、(57.71±3.07)%和(75.50±3.93)%,显著高于对照组(0.96±0.17)%,差异有统计学意义,F=8.82,P=0.003;实时荧光定量PCR结果显示,随着鸦胆子油乳注射液的浓度增加,Bcl-2mRNA的表达减少,Bax mRNA的表达量逐渐上升,与对照组比较差异有统计学意义,P<0.01。结论:鸦胆子油乳能够抑制Hep-2细胞的增殖并诱导其凋亡,作用呈剂量和时间依赖性。

OBJECTIVE：To explore the effect of Brucea javanica oil emulsionon on the growth of human laryngeal Hep-2cells.METHODS：MTT assay was used to observe the proliferation of human laryngeal carcinoma Hep-2cells treated with various concentrations of Brucea javanica oil emulsion at different times.Morphologic changes of Hep-2cells was observed by inverted microscopy after treated with Brucea javanica oil emulsion;Flow cytimetry（FCM）and Real-time quantitative PCR was applied to measure the apoptosis rate and the expression of Bcl-2and Bax mRNA in the Hep-2cells treated with Brucea javanica oil emulsion at 0.2,0.4,0.6g/L for different time.RESULTS：Brucea javanica oil emulsion inhibited growth of Hep-2cells in dose-dependent and time-dependent manner（P0.01）.Typical morphological changes of apoptotic cells,cellular rounding-up,cytoplasmic contraction,chromatin condensation and particularly apoptotic body,the main morphological characteristic of apoptpsis were observed by reserved microscopy.Inhibited cell proliferation and induced death were in a dose-dependent manner,with apoptotic rate of（42.58±4.36）%,（57.71±3.07）% and（75.50±3.93）% accordingly.There existed a statistically significant difference between treatment and control（F=8.82,P=0.003）.Real-time quantitative PCR showed that Brucea javanica oil emulsion could reduce the Bcl-2gene expression and increase gene expression of Bax（P0.01）.CONCLUSION：Brucea javanica oil emulsion has obvious induction of apoptosis on human laryngeal cancer Hep-2cells in a dose-dependent and time-dependent manner.