›› 2012, Vol. 24 ›› Issue (3): 213-216.doi: 10.3969/j.issn.1004-616x.2011.03.011

• 论著 • 上一篇    下一篇

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

靳翠红,巫生文,逯晓波,刘秋芳,蔡 原,肖 云   

  1. 中国医科大学公共卫生学院毒理教研室,辽宁 沈阳 110001
  • 收稿日期:2011-09-02 修回日期:2011-12-16 出版日期:2012-05-30 发布日期:2012-05-30
  • 通讯作者: 蔡 原

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

JIN Cui-hong,WU Sheng-wen,LU Xiao-bo,LIU Qiu-fang,CAI Yuan,XIAO Yun   

  1. Department of Toxicology, School of Public Health, China Medical University, Shenyang 110001, Liaoning, China
  • Received:2011-09-02 Revised:2011-12-16 Online:2012-05-30 Published:2012-05-30
  • Contact: CAI Yuan

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 (conditioned medium,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-效应关系 (P<0.05)。4 mg/L ACTD作用1 h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。

关键词: 放线菌素D, 条件培养液, 旁观者效应, 染色体畸变

Abstract: 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.

Key words: actinomycin D, conditioned medium, bystander effect, choromosome aberration

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