长春新碱对人成骨肉瘤MG63细胞的促凋亡作用及其机制  

Apoptosis inducing effect of vincristine on human osteosarcoma MG63 cells and its mechanisms

作　　者：王洋[1] 苏学今[1] 曲珊珊[1] 娄楠[2] 张海英[1] 史艳芬[1] 李玉林[1] 李荣贵[1] 

机构地区：[1]吉林大学基础医学院病理学教研室,吉林长春130021 [2]吉林大学中日联谊医院骨科,吉林长春130033

出　　处：《吉林大学学报（医学版）》2010年第4期611-615,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30872193);吉林大学留学回国科研启动基金资助课题[科技字(419)号]

摘　　要：目的:观察长春新碱对人成骨肉瘤MG63细胞促凋亡作用,探讨其引起细胞凋亡的分子机制。方法:以0、5、10、20、50、100μg.L-1不同浓度长春新碱处理的MG63细胞,分别经AnnexinV-PI双染和DCFH-DA染色后,应用流式细胞仪检测细胞凋亡和细胞内活性氧化物(ROS)水平,用实时定量RT-PCR分析mRNA表达水平。结果:较低浓度(10μg.L-1)长春新碱即可使MG63细胞凋亡率升高至17.6%(P<0.05),且随着长春新碱浓度的增加细胞凋亡率增加,呈明确的剂量-效应关系;更低浓度(5μg.L-1)长春新碱即可引起细胞内ROS水平明显升高及过氧化氢酶(CAT)基因表达下调(P<0.05)。结论:长春新碱从较低浓度起即有促进细胞凋亡作用,其作用机制是通过下调CAT基因表达,导致细胞内ROS异常蓄积以促进MG63细胞凋亡。Objective To explore the apoptosis inducing effect of vincristine on human osteosarcoma MG63 cells and clarify the molecular mechanisms.Methods The apoptosis and intracellular reactive oxygen species（ROS）level of MG63 cells were determined by flow cytometry,after incubation with Annexin V-PI and DCFH-DA which were explored on different concerntrations of vincristine（0,5,10,20,50,100 μg·L-1）.Real-time quantitative RT-PCR was used to analyze gene expression.Results 10 μg·L-1 vincristine could significantly increase the apoptotic rate of MG63 cells to 17.6%（P〈0.05）,the apoptotic rate was increased with the increasing of concentration of vincristine in a dose-dependent manner;5 μg·L-1 vincristine increased the intracellular ROS level and decreased the expression of CAT gene（P〈0.05）.Conclusion Vincristine can promote the apoptosis of MG63 cells with low concentration,its mechanism is to down-regulate the expresion of CAT gene and induce the increase of ROS to promote apoptosis.

关 键 词：MG63 长春新碱 细胞凋亡 活性氧化物 CAT 

分 类 号：R361.3[医药卫生—病理学]