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目的:探讨中药华蟾素对人宫颈癌Caski细胞生长及凋亡的影响。方法:MTT法检测细胞增殖抑制率,流式细胞术分析细胞周期和鉴定凋亡细胞;琼脂糖凝胶电泳分析DNA片段化;Western Blot分析不同浓度药物处理后细胞胞浆中细胞色素C的含量,荧光免疫组化法观察细胞中活性氧的含量变化,Dioc6染色法检测线粒体膜电位。结果:华蟾素能显著抑制人宫颈癌Caski细胞增殖,抑制效应呈时间和剂量依赖性,其48h的半数抑制浓度值(IC50)为1.68μg/mL;华蟾素可诱导DNA片段化,升高细胞中活性氧(ROS)含量、促进细胞色素C由线粒体向胞浆中释放,降低线粒体膜电位,同时抑制细胞分裂,将细胞阻滞在G2/M期。结论:华蟾素能显著性抑制Caski细胞增殖,其机制可能与增加细胞内活性氧的生成而诱导内源性细胞凋亡和抑制细胞分裂有关,具有用于宫颈癌临床治疗的潜在应用价值。
Objective: To investigate the effect of cinobufacini on the growth and apoptosis of human cervical carcinoma Caski cells. Methods: Cell proliferation inhibition rate was detected by MTT assay, cell cycle was analyzed by flow cytometry and apoptotic cells were identified. DNA fragmentation was analyzed by agarose gel electrophoresis. Cytochrome C content in cytoplasm Fluorescence immunohistochemistry was used to observe the change of reactive oxygen species in the cells. Dioc6 staining was used to detect mitochondrial membrane potential. Results: Cinobufacini significantly inhibited the proliferation of human cervical cancer Caski cells in a time and dose-dependent manner. The IC50 value at 48h was 1.68μg / mL. Cinobufacini induced DNA fragmentation and l High reactive oxygen species (ROS) levels in cells, promote the release of cytochrome C from the mitochondria to the cytoplasm, reduce mitochondrial membrane potential, while inhibiting cell division, the cells arrested in the G2 / M phase. CONCLUSION: Cinobufacini can significantly inhibit the proliferation of Caski cells. The mechanism may be related to the increase of intracellular reactive oxygen species (ROS) generation and the induction of endogenous apoptosis and cell division, which has the potential value of clinical treatment for cervical cancer.