硫利达嗪诱导人卵巢癌细胞凋亡及其机制研究

发布时间：2018-11-05 16:08

【摘要】：目的探讨多巴胺受体阻断剂硫利达嗪(thioridazine)对卵巢癌细胞株SKOV3、A2780增殖、凋亡的影响及其可能的作用机制。方法用不同浓度硫利达嗪(0、5、10、15、20μmol/L)作用SKOV3、A2780细胞,CCK8法检测细胞增殖,流式细胞仪及DAPI染色检测细胞凋亡,DCFH-DA法染色检测ROS水平,彗星实验观察细胞核DNA损伤情况,JC-1染色检测线粒体膜电位变化,Western blot检测P53、Bax、Bcl-2、细胞色素C、active Caspase-3蛋白的表达。结果硫利达嗪可抑制人卵巢癌细胞SKOV3、A2780的增殖,呈一定的剂量效应关系(P0.05),浓度为15μmol/L时,可产生明显增殖抑制效应;流式细胞仪检测显示处理组(15μmol/L硫利达嗪作用24 h)凋亡率明显高于对照组(P0.05);DAPI染色结果:处理组出现明显细胞核固缩、碎裂及凋亡小体;DCFH-DA染色处理组ROS水平升高(P0.05);彗星实验结果提示处理组出现明显的DNA损伤;JC-1染色发现处理组线粒体膜电位较对照组下降(P0.05);Western blot实验发现处理组P53、Bax、胞质细胞色素C、active Caspase-3表达上调,Bcl-2、线粒体细胞色素C表达下调。结论硫利达嗪可能通过诱导细胞内ROS升高损伤DNA,激活线粒体凋亡途径而诱导人卵巢癌SKOV3、A2780细胞凋亡。
[Abstract]:Objective to investigate the effect of (thioridazine), a dopamine receptor blocker, on the proliferation and apoptosis of ovarian cancer cell line SKOV3,A2780 and its possible mechanism. Methods SKOV3,A2780 cells were treated with different concentrations of thiridazide (0 0101010 ~ 1520 渭 mol/L). CCK8 assay was used to detect cell proliferation, flow cytometry and DAPI staining were used to detect cell apoptosis, and DCFH-DA staining was used to detect the level of ROS. The damage of nuclear DNA was observed by comet assay. Mitochondrial membrane potential was detected by JC-1 staining., Western blot was used to detect the expression of P53 Bax-Bcl-2 and cytochrome active Caspase-3 protein. Results the proliferation of human ovarian cancer cell line SKOV3,A2780 was inhibited by thiridamine in a dose-dependent manner (P0.05). When the concentration was 15 渭 mol/L, the proliferation inhibition effect was obvious. Flow cytometry analysis showed that the apoptotic rate of the treatment group (15 渭 mol/L tiridamine for 24 h) was significantly higher than that of the control group (P0.05); DAPI staining results: the treatment group showed obvious nuclear pyknosis, fragmentation and apoptotic bodies; The level of ROS in DCFH-DA staining group was higher than that in control group (P0.05); Comet assay showed obvious DNA damage in treatment group; JC-1 staining showed that mitochondrial membrane potential in treatment group was lower than that in control group (P0.05). Western blot assay showed that the expression of cytochrome C active Caspase-3 in cytochrome C was up-regulated and the expression of cytochrome C in mitochondria of Bcl-2, was down-regulated in the treatment group. Conclusion tiridazide may induce apoptosis of human ovarian cancer SKOV3,A2780 cells by inducing increased DNA, damage and mitochondrial apoptosis in human ovarian cancer SKOV3,A2780 cells.
【作者单位】：重庆医科大学附属第二医院妇产科;重庆医科大学附属第二医院康复科;
【基金】：国家自然科学基金面上项目(81172492) 重庆市科委重点项目(CSTC2012JJB10030)~~
【分类号】：R737.31

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