不同剂量γ射线对人外周血miRNA的表达影响

Effects of the expression of miRNA in human peripheral blood after different doses of gamma rays

目的利用高通量miRNA测序和生物信息学技术,筛选人外周血中辐射敏感的miRNA分子,为辐射损伤提供早期诊断指标。方法采集健康成年男性的外周血,给予0.2 Gy和2.0 Gyγ射线照射,于照射后6 h提取总RNA,应用高通量miRNA测序手段获得差异的miRNA,qRT-PCR方法验证部分差异的miRNA,通过mirdbV6和Target Scan7.1数据库预测共同差异miRNA的靶基因,并进行KEGG生物信息学分析。结果人外周血经不同剂量γ射线照射后6 h,0.2 Gy组差异miRNA有10个,2个表达上调,8个表达下调;2.0 Gy作用后共鉴定出9个表达上调、12个表达下调miRNA分子,差异有统计学意义(P<0.05)。其中有5个miRNA在2个剂量组均发生显著变化。RTPCR实验结果表明有4个miRNA,miR-23c、miR-1287-5p、miR-219a-3p、miR-320d与测序结果一致。生物信息学结果表明,在2个剂量组均差异表达的miRNA可能通过参与调控MARK、RAS、P53、RIG-I等信号通路影响细胞增殖、凋亡、DNA损伤修复及免疫调控。结论miR-23c、miR-1287-5p、miR-219a-3p、miR-320d分子有望成为辐射损伤新的血液标志物。

Objective In order to provide early diagnostic indicators for radiation damage,high throughput miRNA sequencing and bioinformatics technology were used to screen radiation-sensitive miRNA in human peripheral blood.Methods The peripheral blood of healthy adult males was given 0.2 Gy and 2.0 Gy gamma rays,and total RNA was extracted at 6 hours after irradiation.Differential miRNA was obtained by high-throughput miRNA sequencing technology,and partially differential miRNA was verified by qRT-PCR.Target genes of common differential miRNA were predicted by mirdbV6 and Target Scan7.1 database,and bioinformatics analysis was analyzed using KEGG.Results The number of differential miRNAs in human peripheral blood was different with at 6 h after different doses radiation.There were 10 differential miRNAs in 0.2 Gy group,and 2 were up-regulated,and 8 were down-regulated.In 2.0 Gy group,there were 9 up-regulated and 12 down-regulated miRNAs.And the differences of both two groups were statistically significant(P<0.05).There were 5 miRNAs showed significant changes in both groups.RT-PCR results showed that there were 4 miRNAs,including mir-23 c,mir-1287-5 p,mir-219 a-3 p and mir-320 d,which were consistent with the sequencing results.Bioinformatics results showed that common differential miRNAs may affect cell proliferation,cell apoptosis,DNA damage repair and immune regulation by regulating the MARK,RAS,P53,RIG-I and other signaling pathways.Conclusion The mir-23 c,mir-1287-5 p,mir-219 a-3 p and mir-320 d are expected to become new blood makers of radiation injury.