摘要
目的:研究长期高剂量苯并(a)芘[B(a)P染毒对小鼠肺组织miRNAs表达谱的影响,探讨miRNAs在B(a)P健康损害过程中的作用。方法:40只ICR小鼠随机分为对照组和染毒组,每组20只,雌雄各半。经口灌胃给予小鼠50 mg/kg B(a)P,每周2次,持续8周,对照组同时给予等量的橄榄油,染毒结束后继续饲养8周。取肺脏组织,提取总RNA,采用SOliD高通量测序技术检测miRNAs表达谱,进行miRNAs差异表达分析,并用real time-PCR验证miRNAs表达,TargetScan,miRanda及picTar软件预测miRNAs可能调控的靶基因。结果:绝大部分miRNAs在肺组织的表达信号强度较低。与对照组相比,B(a)P染毒组小鼠肺组织中共有109个miRNAs发生差异表达,其中50个miRNAs表达上调,59个miRNAs表达下调。上调幅度最大的为7.43倍,下调幅度最大的为40.63倍。为验证测序的结果,取下调较为明显的miR-20b生行real time-PCR分析,结果显示与测序结果相一致,靶基因预测显示miR-20b可能调节与细胞增殖、周期、凋亡及肿瘤发生相关的蛋白。结论:长期高剂量B(a)P染毒可引起小鼠肺组织miRNAs表达产生特异性改变,差异表达miRNAs可能在B(a)P致机体健康损害过程中起着重要作用。
OBJECTIVE：To study the effect of high-dose long-term benzo（a）pyrene[B（a）P]exposure on miRNAs expression profile in lung tissues of mice and analyze the potential mechanism of miRNAs on the B（a）P-induced health damage.METHODS：40 ICR mice were divided into control group and exposure groups randomly.Each group included 10 male and 10 female mice.Exposure group mice were treated with 50 mg/kg B（a）P（twice a week） for 8 weeks by intragastric administration.Control mice were treated with same volume of olive oil solvent.Then,all mice were fed for another 8 weeks without exposure.The total RNA was isolated from mice lung tissues.The miRNAs expression profiles were evaluated by SOliD high-throughput sequencing technique and the difference expression of miRNAs were analyzed. Real time-PCR was used to confirm the miRNAs expression.The potential targets of miRNAs were calculated by TargetScan,miRanda and picTar software.RESULTS：Most of miRNAs were found at low expression levels in lung tissue.Compared with control mice,109 miRNAs expression changed significantly in lung tissue of B（a）P exposure mice. Of these 109 miRNAs,50 were up-regulated and 59 were down-regulated,the expression changes were 7.43-fold and 40.63-fold,respectively.miR-20b was analyzed by real time-PCR to verify the sequencing.The result of real time-PCR was consistent with sequencing.The potential targeted proteins of miR-20b were involved in cell proliferation,cell cycle,apoptosis and oncogenesis.CONCLUSION：High-dose long-term B（a）P exposure could specifically alter miRNAs expression profiles.The differential expression of miRNAs may play an important role for the B（a）P-induced health damage.
出处
《癌变．畸变．突变》
CAS
CSCD
2012年第2期86-90,共5页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金(30700650)
江苏省自然科学基金(BK2007565)
高校博士点专项(20070286070)
东南大学优秀青年教师项目(2008)
