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目的研究manumycin抑制Tca8113细胞的生长和诱导细胞凋亡的作用及机制。方法细胞毒作用以MTT法测定;凋亡细胞形态以Hoechst33258染色后荧光显微镜观察;凋亡率的检测以AnnexinV染色,流式细胞仪检测;细胞内活性氧(ROS)和线粒体跨膜电位(ΔΨm)分别用DCFH-DA和DiOC6荧光探针标记,流式细胞仪检测;蛋白质定量检测以Westernblot法。结果manumycin浓度依赖性抑制Tca8113细胞的生长,IC50为(11.33±0.63)μmol.L-1;manumycin可浓度和时间依赖性诱导Tca8113细胞的凋亡,过氧化物清除剂N-乙酰半胱氨酸能清除manumycin介导的细胞活性氧的增加,抑制线粒体跨膜电位降低及细胞凋亡。结论manumycin体外显著抑制舌鳞癌Tca8113的生长及诱导细胞凋亡,其机制可能通过激活线粒体依赖性凋亡通路有关。
Objective To study the effect and mechanism of manumycin on inhibiting Tca8113 cell growth and inducing apoptosis. Methods The cytotoxicity was determined by MTT assay. The morphology of apoptotic cells was observed by fluorescence microscopy with Hoechst33258 staining. The apoptosis rate was detected by Annexin V staining and flow cytometry. The intracellular reactive oxygen species (ROS) and mitochondrial transmembrane potential (ΔΨm ) Were labeled with DCFH-DA and DiOC6 fluorescent probes, respectively, and detected by flow cytometry. The protein was detected by Western blot. Results Manumycin inhibited the growth of Tca8113 cells in a concentration-dependent manner with an IC50 of (11.33 ± 0.63) μmol·L-1. Manumycin induced the apoptosis of Tca8113 cells in a concentration-dependent and time-dependent manner. The activity of peroxide scavenger N-acetylcysteine Can eliminate manumycin-mediated increase of reactive oxygen species, inhibit mitochondrial transmembrane potential and apoptosis. Conclusion Manumycin can significantly inhibit the growth and induce the apoptosis of Tca8113 in tongue squamous cell carcinoma in vitro, which may be related to the activation of mitochondria-dependent apoptotic pathway.