放线菌素D对仓鼠成纤维细胞V79靶细胞和旁观者细胞活力和染色体畸变的影响  

Effect of actinomycin D on cell viability and chromosome aberration in hamster fibroblast V79 target and bystander cells

作　　者：靳翠红[1] 巫生文[1] 逯晓波[1] 刘秋芳[1] 蔡原[1] 肖云[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：《癌变．畸变．突变》2012年第3期213-216,共4页Carcinogenesis,Teratogenesis & Mutagenesis

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)对中国仓鼠成纤维细胞V79靶细胞和旁观者细胞的细胞活力和染色体畸变的影响,以确定ACTD能否诱导旁观者效应的发生。方法:用不同剂量ACTD(0、0.25、0.5、1.0、2.0、4.0和8.0 mg/L)处理V79靶细胞1 h,并选择其中4 mg/L ACTD作用1 h开始计时,分别在第4、8、12和24 h时取靶细胞去细胞培养液(conditionedmedium,CM),培养正常细胞24 h后,进行旁观者效应的观察。用MTT法测定靶细胞和旁观者细胞活力;并对靶细胞和旁观者细胞进行染色体畸变分析。结果:随着ACTD剂量的增加,靶细胞的活力下降,存在明显的剂量一效应关系(P<0.05)。4 mg/L ACTD作用1h时细胞活力接近50%且稳定。旁观者细胞活力在用4～12 h CM培养时逐渐增加(P<0.05),用24 h CM时又下降(P<0.05),4 h CM诱导旁观者细胞损伤的作用最强。靶细胞的染色体畸变率随ACTD剂量的增加而增高,具有剂量-效应关系(P<0.05)。旁观者细胞染色体畸变以4 h CM诱导的作用最强,随着加入CM时段的延后,损伤逐渐减轻(P<0.05),在24 h CM又趋严重(P<0.05);旁观者细胞与靶细胞染色体畸变类型相似,以断裂为主,伴有少量环形染色体、碎片和双着丝粒染色体。结论:ACTD可以诱导旁观者效应的发生,染色体畸变在靶细胞和旁观者细胞均以断裂为主。OBJECTIVE：To observe the effect of actinomycin D（ACTD） and conditioned medium on cell viability and chromosome aberration in hamster fibroblast V79 cells to determine whether actinomycin D could induce bystander effect.METHODS：V79 cells were treated with ACTD for 1 h at different doses（0,0.25,0.5,1.0,2.0, 4.0 and 8.0 mg/L）.The 4 mg/L ACTD-conditioned medium（CM） was collected at 4,8,12 and 24 h after treatment for 1 h to culture bystander cells to observe the bystander effect.Cell viability was evaluated by MTT assay;chromosome damage was detected using the conventional analysis of chromosomal aberrations.RESULTS：Cell viability of ACTD- treated V79 cells was reduced in a dose-dependent manner（P〈0.05）,was close to 50 percent and stable when ACTD was at 4 mg/L for 1 h.Cell viability of CM-treated bystander cells increased significantly in a time-dependent manner（P〈0.05） till a rebound in 24 h CM（P〈0.05）.The bystander effect in the 4 h CM was the strongest.Chromosome aberration rate in ACTD-treated cells increased significantly in a ACTD dose-dependent manner（P〈0.05）.Chromosome aberration rate in CM-treated bystander cells decreased significantly in a time-dependent manner（P〈0.05） till a rebound in 24 h CM,with the 4 h CM being the highest.Type of chromosome aberration in bystander cells was mainly breakages along with ring chromosome,fragmented and dicentric chromosome similar to exposed cells.CONCLUSION：ACTD could induce bystander effect in V79 cells.Chromosome aberration type consisted of mainly breakages in both ACTD-treated cells and CM-treated bystander cells.

关 键 词：放线菌素D 条件培养液 旁观者效应 染色体畸变 

分 类 号：R114[医药卫生—卫生毒理学]