四氧嘧啶对人神经瘤SK-N-SH细胞增殖、凋亡的影响及机制探讨  

Effect and mechnism of alloxan on multiplication and apoptosis in human neuroblastma cells

作　　者：丁楠[1] 王景景[1] 熊慧[1] 禇玉晶 高蕊[1] 丁玲[1] 戎凯[1] 赵纲[2] 邓艳秋[1] 

机构地区：[1]天津医科大学基础医学院,天津300070 [2]天津医科大学肿瘤医院

出　　处：《山东医药》2012年第2期1-3,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30973156);美国NIHFIRCA国际合作项目(R03TW008123-01A1);天津市高等学校科技发展基金项目(20070203)

摘　　要：目的观察四氧嘧啶对人神经瘤SK-N-SH细胞增殖和凋亡的影响,并探讨其机制。方法将人神经瘤SK-N-SH细胞分为两组,对照组常规培养12 h,观察组分别加不同浓度的四氧嘧啶培养12 h。用MTT法测细胞吸光度值观察细胞增殖情况,用PI/Hoechst染色法观察细胞凋亡,用蛋白免疫印迹法检测细胞中糖基化和磷酸化的神经丝蛋白。结果对照组细胞吸光度值为0.43±0.01,观察组加入2、4、6、8、10 mM四氧嘧啶作用12 h,细胞吸光度值分别为0.39±0.01、0.35±0.02、0.30±0.01、0.26±0.03、0.21±0.02;观察组与对照组比较,P均<0.05;观察组不同浓度间比较,P均<0.05。对照组细胞凋亡率为5%±1%,观察组加入2、4、6 mM四氧嘧啶作用12 h,细胞凋亡率分别为10%±1%、18%±2%、29%±1%;观察组与对照组比较,P均<0.05;观察组不同浓度间比较,P均<0.05。对照组细胞中磷酸化、糖基化神经丝蛋白OD值为1.00±0.20、1.00±0.20,观察组加入2、4、6 mM四氧嘧啶作用12 h,细胞中磷酸化、糖基化神经丝蛋白OD值分别为1.21±0.42和0.95±0.21、1.32±0.51和0.78±0.32、1.51±0.30和0.75±0.24;观察组与对照组比较,P均<0.05;观察组不同浓度间比较,P均<0.05。结论四氧嘧啶可抑制SK-N-SH细胞增殖,并诱导其凋亡;可能与神经丝蛋白的糖基化降低、磷酸化增加有关。Objective To observe the effect of alloxan on multiplication and apoptosis in human neuroblastma SK-N- SH cells, and explore the mechanism. Methods Human neuroblastoma SK-N-SH cells were divided into two groups, the control group were cultured routinely for 12 h, the experiment group were cultured with different concentration of alloxan for 12 h. MTT assey was used to observe the multiplication of cells. Pl/Hoechst staining was used to observe cell apoptosis. Western blot was used to detect O-Glycosalytion and phosphorylation of cell neurofilament proteins. Results The OD value of the control group was 0.43 ±0.01, the OD values of the experiment with 2,4,6,8,10 mM of Alloxan for 12 h were 0.39 ± 0.01,0.35 ± 0.02,0.30 ± 0.01, 0.26 ± 0.03, 0.21 ± 0.02 individually （ all P 〈 0.05 ）. Apoptosis rate of the control group was 5% ± 1%, apoptosis rate of the experiment group with 2,4,6 mM of alloxan for 12 h were 10% ± 1% ,18% ± 2% ,29% ± 1% individually（ all P 〈 0.05）. The OD values of phosphorylation and O-Glycosylation of the control group were 1.00 ±0.20 and 1.00 ±0.20 individually, those of the experiment group with 2,4,6,8,10 mM of alloxan for 12 h were 1.21 ±0.42,0.95 ±0.21,1.32 ±0.51,0.78 ±0.32,1.51 ±0.30,0.75 ±0.24 individually（ all P 〈0.05）. Conclusion Alloxan can inhibit the multiplication of the SK-N-SH cells, and induce apoptosis. It maybe relative to the desease of O-Glycosylation and the increase of phosphorylation of neurofilament proteins.

关 键 词：四氧嘧啶 神经丝蛋白 蛋白糖基化 蛋白磷酸化 细胞增殖 细胞凋亡 

分 类 号：R743.1[医药卫生—神经病学与精神病学]