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【目的】观察牛蒡子苷(ARC)与牛蒡子苷元(ARG)对人前列腺癌PC3细胞增殖的影响,并探讨其相关机制。【方法】采用不同浓度的ARC与ARG作用于PC3细胞,四甲基偶氮唑盐(MTT)法检测其对细胞增殖的影响;瑞姬氏染色观察用药前后细胞形态变化;Annexin V-异硫氰酸荧光素-碘化丙啶(FITC/PI)双染结合流式细胞术检测细胞凋亡或坏死情况;Western-blot法检测凋亡相关蛋白Bcl-2、Bax和Caspase 3的表达情况。【结果】ARC与ARG均能抑制PC3细胞的增殖,此抑制作用具有时间和浓度依赖性,两药物作用48 h组细胞存活率均显著低于24 h组(P<0.01);ARC与ARG处理组的细胞形态变化表现为细胞膜回缩,细胞质减少,胞膜紧贴胞核,胞液纤维网状结构;流式细胞术检测发现:与空白对照组比较,ARC组与ARG组可显著增加Annexin V-FITC/PI双染阳性率(P<0.05或P<0.01),PI单染阳性率在浓度为20μmol/L和5μmol/L时也显著增加(P<0.01),但Annexin V-FITC单染阳性率均无显著变化(P>0.05)。Western-blot分析结果显示:ARC或ARG作用细胞48 h时,可显著降低Bcl-2表达水平(P<0.01),但对Bax和Caspase-3蛋白的表达无显著影响(P>0.05)。【结论】ARC与ARG可诱导PC3细胞发生非凋亡性死亡,其作用机制可能与诱导Bcl-2表达下调相关。
【Objective】 To observe the effects of arctiin (ARC) and arctigenin (ARG) on the proliferation of human prostate cancer PC3 cells and to explore its mechanism. 【Methods】 PC3 cells were treated with different concentrations of ARC and ARG. The effects of ARC and ARG on the proliferation of PC3 cells were detected by MTT assay. The changes of cell morphology were observed before and after treatment with Reiki’s stain. Cell apoptosis or necrosis was detected by double staining of FITC / PI and flow cytometry. The expressions of Bcl-2, Bax and Caspase 3 were detected by Western-blot. 【Results】 Both ARC and ARG inhibited the proliferation of PC3 cells in a time and concentration-dependent manner. The survival rates of the two drugs at 48 h were significantly lower than those at 24 h (P <0.01) The morphological changes of the cells showed cell membrane retraction, cytoplasm reduction, cell membrane close to the nucleus, cytosol network structure; flow cytometry found: compared with the blank control group, ARC group and ARG group can significantly increase Annexin (P <0.05 or P <0.01). The positive rate of PI staining was also significantly increased at the concentrations of 20 μmol / L and 5 μmol / L (P <0.01), but the positive rate of Annexin V-FITC There was no significant difference in the positive staining rate (P> 0.05). Western-blot analysis showed that ARC or ARG-treated cells could significantly decrease the expression of Bcl-2 at 48 h (P <0.01), but had no effect on the expression of Bax and Caspase-3 (P> 0.05). 【Conclusion】 ARC and ARG can induce non-apoptotic PC3 cell death and its mechanism may be related to the down-regulation of Bcl-2 expression.