ACTD处理对旁观者V79细胞ROS和细胞色素C分布的影响  

Effect of conditioned medium from actinomycin D-treated cells on ROS levels and distribution of cytochrome C in V79 bystander cells

作　　者：靳翠红[1] 刘秋芳[1] 巫生文[1] 逯晓波[1] 刘洋[1] 蔡原[1] 

机构地区：[1]中国医科大学公共卫生学院毒理教研室,辽宁沈阳110001

出　　处：《毒理学杂志》2012年第6期399-401,共3页Journal of Toxicology

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V79靶细胞获得的条件培养液(conditioned medium,CM)对V79旁观者细胞ROS水平和细胞色素C(Cytochorome C,Cyt C)分布的影响,以研究ACTD诱导旁观者效应发生的可能机制。方法 4.0 mg/L ACTD处理V79靶细胞1 h,磷酸盐缓冲液(PBS)洗3次,加入新鲜培养液开始计时,分别在4、8、12和24 h获取靶细胞条件培养液CM。用不同时段CM培养正常V79细胞24 h后,流式细胞仪观察旁观者细胞ROS水平;比色法测定胞浆和线粒体细胞色素C含量。结果 CM处理组旁观者细胞ROS水平均高于对照组,随着时段的延后,ROS水平逐渐降低(P<0.01),24 h又增加(P<0.01)。对照组线粒体内Cyt C含量最高,在各CM处理组旁观者细胞线粒体Cyt C含量均有不同程度下降,4 h CM组含量最低,随着时段延后,线粒体Cyt C含量增加,但没达到对照组水平,24 h又趋下降。胞浆内Cyt C含量变化趋势与之相反,对照组Cyt C含量最低,CM处理组胞浆Cyt C含量增加,4 h最高随后下降,24 h又明显增加。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ROS的产生,并造成细胞色素C从线粒体释放而引起的。Objective To observe the effect of conditioned medium （CM） obtained from actinomycin D （ACTD）-treated cells on ROS levels and distribution of cytochrome c to explore the underlying mechanism that ACTD induces bystander effect in V79 cells. Methods V79 cells were administrated with 4. 0 mg/L ACTD for 1 h then washed with PBS 3 times. Immediately fresh mediums were supplemented and counted as 0 h. From then on, different period conditioned mediums were collected at 4, 8, 12 and 24 h to culture bystander cells for 24 h to observe the bystander effect. ROS levels were detected by flow cytometry; colorimetric method was used to determine the distribution of cytochrome c in mitochondria and cytoplasm. Results ROS levels in CM-treated bystander cells were higher than those in control CM. With the time of CM going, ROS levels decreased till 12 h whereas increased in 24 h CM group （P 〈 0.01 ）. In mitochondria, the content of cytochrome c decreased in CM-treated bystander cells compared to the control. With the time of CM going, cytochrome c contents increased till 12 h whereas decreased in 24 h CM group （P 〈 0. 01 ）. On the contrary, the content of cytochrome c increased in cytoplasm of CM- treated bystander cells compared to the control. With the time of CM going, cytochrome c content decreased till 12 h whereas increased in 24 h CM group （P 〈 0. O1 ）. Conclusion ACTD could induce bystander effect through the conditioned medium by inducing ROS generation and rendering cytochrome c release from mitochondria into cytoplasm in bystander V79 cells.

关 键 词：放线菌素D 条件培养液 ROS 细胞色素C 

分 类 号：R114[医药卫生—卫生毒理学]