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目的探讨紫外线灭活仙台病毒致人胶质瘤LN229细胞凋亡作用及其机制。方法用不同剂量灭活病毒感染LN229细胞,24h后,MTT法检测灭活病毒对细胞增殖的影响;流式细胞仪检测细胞凋亡情况;分光光度法检测caspase活性;Western blotting检测凋亡相关蛋白的表达水平。结果 MTT检测显示灭活仙台病毒能够剂量依赖性抑制LN229细胞增殖;流式细胞仪检测显示灭活病毒组细胞凋亡率呈剂量依赖性升高;caspase活性测定显示灭活病毒组细胞中caspase-3,-8和-9蛋白活性呈剂量依赖性增加;Western blotting结果显示,随着灭活病毒滴度增加,细胞中Bax、细胞色素c、Fas、FasL表达水平升高、而Bcl-2、caspase-8前体、caspase-9前体和caspase-3前体蛋白表达下降。结论灭活仙台病毒能够诱导LN229细胞剂量依赖性凋亡,其机制与线粒体途径和死亡受体途径相关。
Objective To investigate the apoptosis of human glioma LN229 cells induced by ultraviolet (UV) inactivated Sendai virus and its mechanism. Methods LN229 cells were infected with different doses of inactivated virus, and the effect of inactivated virus on cell proliferation was detected by MTT assay. The apoptosis of cells was detected by flow cytometry. The caspase activity was detected by spectrophotometry. The level of expression. Results MTT assay showed that inactivation of Sendai virus could inhibit the proliferation of LN229 cells in a dose-dependent manner. The apoptosis rate of LN229 cells was increased in a dose-dependent manner by flow cytometry. The activity of caspase- 3, -8 and -9 protein in a dose-dependent manner. Western blotting showed that the expression of Bax, cytochrome c, Fas and FasL increased with the increase of inactivated virus titer, while the expression of Bcl-2, Caspase-8 precursor, caspase-9 precursor and caspase-3 precursor protein expression decreased. Conclusion Inactivation of Sendai virus can induce dose-dependent apoptosis of LN229 cells, and its mechanism is related to mitochondrial pathway and death receptor pathway.