异紫堇定碱对人宫颈癌Siha细胞增殖影响的实验研究

Effects of isocorydione on cell proliferation in SiHa human cervical carcinoma cell lines

目的:探讨异紫堇定碱(Isocorydione)对人宫颈癌Siha细胞增殖的影响。方法:以100、200、400、800、1 200μmol/L浓度的异紫堇定碱对人宫颈癌Siha细胞体外干预,分别作用24、48、72 h后,采用MTT法检测异紫堇定碱对人宫颈癌Siha细胞生长增殖的作用;流式细胞仪检测异紫堇定碱对人宫颈癌Siha细胞周期的影响;结合Hoechst染色观察处理后Siha细胞核微形态的变化;Western Blot法检测异紫堇定碱对人宫颈癌Siha细胞作用后凋亡相关蛋白Bax、Bcl-2、Caspase-3表达的影响。结果:MTT结果显示不同浓度的异紫堇定碱作用于人宫颈癌Siha细胞后具有明显的抑制其增殖的作用,呈剂量-时间依赖关系(P<0.05);流式细胞仪检测结果显示400μmol/L的异紫堇定碱作用于人宫颈癌Siha细胞后,细胞周期发生明显改变,其中G1和S期细胞比例变化不明显,但G2期细胞明显减少(P<0.05);Hoechst染色结果显示,与对照组比较,400μmol/L的异紫堇定碱作用于Siha细胞48 h后形态缩小、细胞核固缩,出现核碎裂形成凋亡小体;Western Blot结果显示400μmol/L的异紫堇定碱分别作用于人宫颈癌Siha细胞24、48、72 h后,其Bax蛋白表达增加,而Bcl-2蛋白表达逐渐减少,Bax/Bcl-2比值增加,Caspase-3蛋白表达逐渐增多。结论:异紫堇定碱对人宫颈癌Siha细胞增殖具有明显的抑制作用,其促使细胞发生凋亡行为可能与线粒体凋亡途径的蛋白有关。

Objective：This paper aimed to investigate the effects of isocorydinone on cell proliferation in SiHa human cervical carcinoma cell lines. Methods：Different concentrations of isocorydione （100, 200, 400, 800, and 1200 μmol/L） were used to treat SiHa human cervical carcinoma cells in vitro for 24, 48, and 72 h. Methyl thiazolyl tetrazolium （MTT） assays were conducted to determine the inhibitory action of isocorydione. Flow cytometry was performed to detect the cell cycle in SiHa human cervical carcinoma cells af-ter treatment with 400 μmol/L isocorydione. Hoechst 33342 staining was used to observe the micro-morphological changes of SiHa cell nucleus after the treatment. The expression of Bcl-2, Bax, and caspase-3 proteins in cervical carcinoma SiHa cell lines was determined using western blot analysis. Results： MTT assays showed that isocorydione inhibits the proliferation of SiHa cells in a dose- and time-dependent manner （P〈0.05）. The flow cytometry results showed that SiHa cervical carcinoma cells treated with different concen-trations of isocorydione exhibited increased cell cycle. Compared with the control group, Hoechst 33342 staining showed that SiHa cells became narrow, with nuclear pyknosis and fragmentation, and formed an apoptotic body after treatment with 400 μmol/L isocoryd-ione for 48 h. Furthermore, western blot analysis proved that isocorydione significantly inhibited the proliferation of SiHa cell lines, and the expression of Bax protein was increased. By contrast, the expression of Bcl-2 protein decreased gradually. Consequently, the ra-tio of Bax/Bcl-2 increased, as well as the expression of caspase-3 protein. Conclusion：Isocorydione exhibited an overt inhibitory ac-tion on SiHa cells. Isocorydione promoted the occurrence of cell apoptosis, which may be associated with related proteins of mitochon-drial apoptotic pathway.