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目的研究γ射线对人周围血淋巴细胞Tob1基因转录表达的影响。方法对数生长期的淋巴细胞,分别给予1、2、3、4、5、6Gy的60Coγ射线照射,照射后12、24h,以及2Gy照射后4、8、12、24、36、48、72h分别提取总RNA,反转录成cDNA。利用实时荧光定量聚合酶链反应技术,检测各组Tob1基因表达改变。结果2Gy照射后4、24、36h,Tob1 mRNA表达水平升高,均是对照组(未照射组)的1.99倍以上(P<0.05)。1～5Gy范围内照射后24h,Tob1 mRNA表达水平随着剂量的增加而增加差异有统计学意义(P<0.05)。在1～5Gy范围内照射后12h,Tob1 mRNA表达水平和对照组比较差异无显著性(P>0.05);6Gy时其表达水平达最高,为对照组的16.37倍(P<0.05)。结论γ射线照射后人淋巴细胞Tob1基因的表达,在一定剂量范围内的特定时间内呈剂量依赖性上调,Tob1可能会发展为核事故受照射人员的分子生物学剂量标记物。
Objective To study the effects of γ-rays on the transcriptional expression of Tob1 gene in human peripheral blood lymphocytes. Methods The logarithmic growth phase lymphocytes were irradiated with 1, 2, 3, 4, 5 and 6 Gy of 60Coγ ray respectively. After irradiation for 12 and 24 h, and 4, 8, 12, 24, 36, 48 and 72 h Total RNA was extracted and reverse transcribed into cDNA. The real-time fluorescent quantitative polymerase chain reaction technique was used to detect the changes of Tob1 gene expression in each group. Results The mRNA expression of Tob1 at 4, 24, 36 h after 2Gy irradiation were all increased by 1.99 times (P <0.05) in control group (non-irradiated group). The level of Tob1 mRNA increased with the increase of dose at 1 ~ 5 Gy within 24 hours after irradiation. There was significant difference (P <0.05). At 12h after irradiation, Tob1 mRNA expression was not significantly different from that of the control group (P> 0.05). The expression level of Tob1 mRNA reached the highest at 6Gy, which was 16.37 folds higher than that of the control group (P <0.05). Conclusion The expression of Tob1 gene in human lymphocytes after γ-ray irradiation dose-dependently increases within a certain dose range for a certain period of time, and Tob1 may develop into a molecular biological dose marker for irradiated nuclear accidents.