ACTD的CM对旁观者V79细胞Δψm和胞质内细胞色素C的影响  

Effect of ACTD-treated conditioned medium on mitochondria membrane potential and cytochrome C content in bystander V79 cells

作　　者：靳翠红[1] 巫生文[1] 刘秋芳[1] 蔡原[1] 

机构地区：[1]中国医科大学公共卫生学院,辽宁沈阳110001

出　　处：《工业卫生与职业病》2014年第1期18-20,共3页Industrial Health and Occupational Diseases

基　　金：国家自然基金项目资助(30471475)

摘　　要：目的观察放线菌素D(actinomycin D,ACTD)处理中国仓鼠成纤维靶细胞(the Chinese hamster fibroblast V79-C3cell line,V79细胞)获得的条件培养液(conditioned medium,CM)对V79旁观者细胞线粒体膜电位(Δψm)和胞质内细胞色素C(cytochrome C,Cyt C)含量的影响,以研究ACTD诱导旁观者效应发生的可能机制。方法用4.0mg/L ACTD处理V79靶细胞1h,磷酸盐缓冲液(phosphate buffer solution,PBS)漂洗3次,加入新鲜培养液开始计时,分别在第4、8、12和24h获取靶细胞条件培养液CM。不同时段CM培养正常V79细胞24h后,采用罗丹明123(Rhodamine 123,Rh123)荧光染色流式细胞仪法检测旁观者细胞的线粒体膜电位(Δψm);比色法测定胞质内Cyt C的含量。结果各CM处理组旁观者细胞Δψm均比对照组下降(P<0.01)。随着时段的延后,Δψm平均荧光强度在逐渐增加,24h又减少(P<0.01)。胞质内Cyt C含量以对照组最低,CM处理组胞质内Cyt C含量增加(P<0.01),4h最高,随后下降,24h又明显增加(P<0.01)。结论 ACTD诱导旁观者效应可能通过靶细胞的CM影响旁观者细胞线粒体膜电位下降和胞质内Cyt C含量增加,进而引起旁观者细胞凋亡。Objective To observe the effect of conditioned medium（CM）obtained from actinomycin D （ACTD） treated cells on mitochondria membrane potential（Abm）and cytochrome C content in cytoplasm and to explore the potential mechanism of ACTD inducing the bystander effect in Chinese hamster fibroblast V79-- C3 cells. Methods V79 cells were administrated with 4.0 mg/L ACTD for 1 h, and washed with PBS 3 times. Immediately fresh medium were supplemented and counted as 0 h. From then on, different period conditioned medium was collected at 4, 8, 12 and 24 h to culture bystander cells for 24 h and to observe the bystander effect. Immuno staining flow cytometry was used to detect Abm. Colorimetric method was used to determine the content of cytochrome C in cytoplasm. Results Ams in CM--treated bystander cells were lower than those in control CM. With the time of CM prolonged, Abms increased till 12 h whereas decreased in 24 h CM group（P〈0.01）. The content of cytochrome C decreased in cytoplasm of CM--treated bystander cells compared to that in control CM. With the time of CM passed on, cytochrome C contents increased till 12 h whereas decreased in 24 h CM group （P〈0.01）. Conclusions ACTD could induce bystander effect through the conditioned medium destroying Abm and inducing cytochrome C release into cytoplasm in bystander V79 cells.

关 键 词：放线菌素D 条件培养液 线粒体膜电位 细胞色素C 

分 类 号：R114[医药卫生—卫生毒理学]