Survivin抑制剂YM155对视网膜母细胞瘤Y79细胞线粒体凋亡途径的影响

发布时间：2018-07-27 15:59

【摘要】：目的:探讨人生存素(survivin)抑制剂YM155{4,9-二氢-1-(2-甲氧基乙基)-2-甲基-4,9-二氧代-3-(2-吡嗪甲基)-1H-萘并[2,3-d]咪唑摀溴化物}对视网膜母细胞瘤Y79细胞凋亡、线粒体膜电位(△ψ_m)和细胞色素C(Cyt C)的影响,探讨其诱导Y79细胞凋亡的线粒体机制。方法:体外培养Y79细胞,分别以0、0.5、1、2、4、8nmol/L的YM155进行处理,采用CCK-8法和溴脱氧尿嘧啶核苷(bromodeoxyuridine,BrdU)掺入标记法检测YM155对Y79细胞增殖的抑制作用。Y79细胞随机分为4组:对照组、阳性对照组[加入10nmol/L拓扑替康(topotecan)]和低剂量(1 nmol/L)、高剂量(2 nmol/L)YM155组。每组设3个复孔,处理24h。分别采用TUNEL染色和Annexin V-FITC/PI双染法检测凋亡的情况;采用JC-1活细胞染色检测△ψ_m的变化;采用免疫荧光分析检测Cyt C的分布;采用Western blot法检测survivin和线粒体内Cyt C的蛋白表达。结果:与对照组比较,YM155对Y79细胞增殖具有明显抑制作用,并诱导Y79细胞凋亡(P0.05);YM155显著降低Y79细胞的△ψ_m,促进Cyt C由线粒体释放至胞浆,降低线粒体内Cyt C的水平(P0.05)。结论:YM155对Y79细胞增殖具有抑制作用,并诱导细胞凋亡其机制可能是通过线粒体介导的细胞凋亡途径实现的。
[Abstract]:Objective: to investigate the apoptosis of retinoblastoma Y79 cells induced by human survivin (survivin) inhibitor YM155 {4O9-dihydro-1- (2-methoxyethyl) -2-methyl-4-dioxy-9- (2-pyrazinomethyl) -1H-naphthyl [2H- 3-d] imidazolium bromide}. The effects of mitochondrial membrane potential (蠄 M) and cytochrome C (Cyt C) (C (Cyt C) on apoptosis of Y79 cells were investigated. Methods: Y79 cells were cultured in vitro. Y79 cells were treated with YM155 of 0 0. 5, 1, 1, 2, 4 and 4 nmol / L, respectively. The inhibitory effect of YM155 on the proliferation of Y79 cells was detected by CCK-8 assay and bromodeoxyuridine BrdU incorporation method. Y79 cells were randomly divided into 4 groups: control group. Positive control group [10nmol/L topotecan (topotecan)] and low dose (1 nmol/L), high dose (2 nmol/L) YM155 group. Each group was treated with 3 multiple holes for 24 h. TUNEL staining and Annexin V-FITC/PI double staining were used to detect apoptosis; JC-1 living cell staining was used to detect the change of 蠄 m; immunofluorescence assay was used to detect the distribution of Cyt C; Western blot method was used to detect the expression of Cyt C in survivin and mitochondria. Results: compared with the control group, YM155 significantly inhibited the proliferation of Y79 cells, and induced apoptosis of Y79 cells (P0.05). YM155 significantly decreased 蠄 m of Y79 cells, promoted the release of Cyt C from mitochondria to cytoplasm and decreased the level of Cyt C in mitochondria (P0.05). Conclusion the cell proliferation of Y79 cells was inhibited by WYM155, and the mechanism of inducing cell apoptosis may be mediated by mitochondria mediated apoptosis.
【作者单位】：牡丹江医学院红旗医院眼科;牡丹江医学院红旗医院放射科;
【基金】：黑龙江省卫生计生委科研课题(No.JS215H623)
【分类号】：R739.7

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