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目的:观察20(s)-原人参二醇(PPD)对体外培养人宫颈癌Siha细胞中caspase-9和caspase-3转录表达的影响,以及caspase-3的活化形式cleaved caspase-3含量的变化,阐明其诱导Siha细胞凋亡的机制。方法:体外培养人宫颈癌Siha细胞,将其分为阴性对照组(乙醇)和实验组(20μg.L-1 PPD),分别用乙醇和PPD处理体外培养的宫颈癌Siha细胞,48h后流式细胞仪检测细胞的凋亡峰,半定量RT-PCR、Western blotting和免疫细胞化学染色方法检测PPD处理后Siha细胞中caspase-9和caspase-3基因的转录与表达水平。结果:与阴性对照组比较,20μg.L-1 PPD处理48h后,Siha细胞凋亡率增加(P<0.05),Siha细胞中caspase-9与caspase-3转录水平升高(P<0.01),表达上调(P<0.01),caspase-3的活化形式cleaved caspase-3含量增加(P<0.01)。细胞免疫化学检测可见实验组细胞出现棕色颗粒。结论:PPD可促进人宫颈癌Siha细胞凋亡,其机制与激活caspase家族级联反应有关。
OBJECTIVE: To investigate the effect of 20 (s) -protopanaxadiol (PPD) on the transcriptional expression of caspase-9 and caspase-3 in Siha cells cultured in vitro and the change of cleaved caspase-3 in caspase-3 , Clarifying its mechanism of inducing Siha cell apoptosis. Methods: Siha cells were cultured in vitro and divided into negative control group (20μg.L-1 PPD) and experimental group (20μg.L-1 PPD). Siha cells cultured in vitro were treated with ethanol and PPD respectively. After 48 hours, The apoptosis peak of Siha cells were detected by semi-quantitative RT-PCR, Western blotting and immunocytochemical staining. Results: Compared with the negative control group, the apoptotic rates of Siha cells increased (P <0.05) and the caspase-9 and caspase-3 transcriptional levels increased in Siha cells treated with 20μg.L-1 PPD for 48h (P <0.01) (P <0.01). The cleaved caspase-3, an activated form of caspase-3, was increased (P <0.01). Immunocytochemistry showed that the experimental group of cells appear brown particles. Conclusion: PPD can promote the apoptosis of Siha cells in human cervical cancer. The mechanism is related to the caspase family cascade activation.