Lee Eon-Seok, Won Yeo Jin, Kim Byoung-Chul, Park Daeui, Bae Jin-Han, Park Seong-Joon, Noh Sung Jin, Kang Yeong-Rok, Choi Si Ho, Yoon Je-Hyun, Heo Kyu, Yang Kwangmo, Son Tae Gen
Research Center, Dongnam Institute of Radiological and Medical Science, 40 Jwadong-gil, Jangan-eup, Gijang-gun, Busan, 46033, Republic of Korea.
In silico Toxicology Research Center, Korea Insititute of Toxciology, Daejeon 305-343, Republic of Korea.
Sci Rep. 2016 May 26;6:25723. doi: 10.1038/srep25723.
Current evidence indicates that there is a relationship between microRNA (miRNA)-mediated gene silencing and low-dose irradiation (LDIR) responses. Here, alterations of miRNA expression in response to LDIR exposure in male BALB/c mice and three different types of hepatocytes were investigated. The miRNome of the LDIR-exposed mouse spleens (0.01 Gy, 6.5 mGy/h) was analyzed, and the expression of miRNA and mRNA was validated by qRT-PCR. Western blotting, chromatin immunoprecipitation (ChIP), and luciferase assays were also performed to evaluate the interaction between miRNAs and their target genes and to gain insight into the regulation of miRNA expression. The expression of miRNA-193b-3p was down-regulated in the mouse spleen and liver and in various hepatocytes (NCTC, Hepa, and HepG2 cell lines) in response to LDIR. The down-regulation of miR-193b-3p expression was caused by histone deacetylation on the miR-193b-3p promoter in the HepG2 cells irradiated with 0.01 Gy. However, the alteration of histone deacetylation and miR-193b-3p and Rad51 expression in response to LDIR was restored by pretreatment with N-acetyl-cyctein. In conclusion, we provide evidence that miRNA responses to LDIR include the modulation of cellular stress responses and repair mechanisms.
目前的证据表明,微小RNA(miRNA)介导的基因沉默与低剂量辐射(LDIR)反应之间存在关联。在此,我们研究了雄性BALB/c小鼠及三种不同类型肝细胞在LDIR照射后miRNA表达的变化。分析了接受LDIR照射的小鼠脾脏(0.01 Gy,6.5 mGy/h)的微小RNA组,并通过qRT-PCR验证了miRNA和mRNA的表达。还进行了蛋白质免疫印迹、染色质免疫沉淀(ChIP)和荧光素酶测定,以评估miRNA与其靶基因之间的相互作用,并深入了解miRNA表达的调控机制。在小鼠脾脏和肝脏以及各种肝细胞(NCTC、Hepa和HepG2细胞系)中,miRNA-193b-3p的表达在LDIR照射后下调。在用0.01 Gy照射的HepG2细胞中,miR-193b-3p启动子上的组蛋白去乙酰化导致了miR-193b-3p表达的下调。然而,通过N-乙酰半胱氨酸预处理可恢复LDIR照射后组蛋白去乙酰化以及miR-193b-3p和Rad51表达的变化。总之,我们提供的证据表明,miRNA对LDIR的反应包括对细胞应激反应和修复机制的调节。