人质膜型唾液酸酶(Neu3)抑制线粒体途径细胞凋亡  

Mechanisms of the antiapoptosis of mitochondrion of plasma membrane-associated sialidase(Neu3)

作　　者：许莉[1] 孙连坤[2] 禹彬[2] 赵雪俭[2] 李扬[2] 

机构地区：[1]东华大学化学化工与生物工程学院,上海201620 [2]吉林大学基础医学院病理生理学教研室,吉林长春130021

出　　处：《中国实验诊断学》2009年第12期1689-1691,共3页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探讨人质膜型唾液酸酶(Neu3)拮抗前列腺癌线粒体途径细胞凋亡的可能机制。方法建立转染Neu3前列腺癌细胞模型,分别或联合柔红霉素(DNR)与唾液酸酶特异性抑制剂(NeuAC2en)或白细胞介素-6(IL-6)作用;利用DCFH-DA染色法检测细胞ROS水平;JC-1染色法检测细胞线粒体膜电势;分离细胞线粒体和细胞浆检测药物作用下Bcl-xL、Bax变化;结果(1)DCFH-DA法检测ROS变化发现:应用DNR后细胞绿色荧光强度显著增强,NeuAC2en可以增强DNR这种药物作用;IL-6则可以拮抗其作用;(2)JC-1检测细胞线粒体膜电势发现:三组不同细胞中,与未加药组相比应用DNR细胞其线粒体膜电势明显降低,NeuAC2en可以增强DNR这种药物作用,IL-6则可以拮抗其作用;与对照组相比,应用DNR后PC-3-C1-Neu3细胞膜电势比较稳定(P<0.05);(3)WB法发现:与对照组相比相同处理条件下PC-3-C1-Neu3细胞浆和线粒体中Bcl-xL表达水平均较高,而Bax表达则水平较低,细胞膜上绿色荧光表达明显增多。结论Neu3通过稳定线粒体膜电势和抑制活性氧的产生抵抗细胞凋亡;Neu3可能通过改变Bcl-2家族蛋白中相关因子与其底物GD3含量抵抗细胞凋亡。Objective To investigate the impossible mechanism of the impact of Neu3 on mitochondrial pathways of apoptosis in prostate cancer cells.Methods The wild type pc-3 cells,the pc-3 cells transfected with Neu3 and the mock vector were treated by DNR alone or in DNR combine with IL-6 and NeuAC2en.The level of intracellular ROS of PC-3,PC-3-C1 and PC-3-C1-Neu3 cells treated by drugs was examined with DCFH-DA staining.The mitochondrial membrane potential change of PC-3-C1 and PC-3-C1-Neu3 cells treated by drugs was investigated with JC-1 staining.The level of Bcl-xL and Bax in cytosome and motochondria of PC-3-C1 and PC3-C1-Neu3 cells treated by drugs was examined respectively through western blot.Results PC-3 cells treated by DNR demonstrated stronger green fluorescence contrasting control group,and PC-3 cells treated by IL-6 and NeuAC2en alone.Strongest green fluorescence was produced through DNR and NeuAC2en coordination.Under identical treatment condition,fluorescence intensity of PC-3-C1-Neu3 was lower than that of PC-3-C1 Cells.The mitochondrial potential of PC-3-C1 and PC3-C1-Neu3 cells treated by DNR was reduced than that of respective control group.Mitochondria membrane potential of PC-3-C1-Neu3 cells was higher than PC-3-C1 cells under the identical treatment conditions.NeuAC2en can enforce DNR effects.Compared with control group respectively,the protein expression of Bcl-xL and Bax decreased in cytoplasm and increased in mitochondria of PC-3-C1 and PC-3-C1-Neu3 treated by DNR.The level of Bcl-xL protein was higher and the level of Bax protein was lower in both cytoplasm and mitochondria of PC-3-C1-Neu3 cells than those of PC-3-C1 cells under identical treatment conditions.PC-3 cell membrane presented green fluorescence;The cells treated by IL-6 expressing green fluorescence was less.Conclusion Neu3 could stabilize the mitochondrial membrane potential,repress the burst of ROS and reduce the proportion of Bcl-xL/Bax to resist apoptosis,but Neu3 have no effect on translocation of Bax and Bcl-xL.

关 键 词：人质膜型唾液酸酶 细胞凋亡 前列腺癌 

分 类 号：R737.25[医药卫生—肿瘤]