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目的:研究受γ射线照射后人外周血中POLH mRNA的表达变化,为确立POLH mRNA作为新的辐射损伤分子生物标志物提供实验依据。方法:采取3名健康成人的外周血,利用~(60)Coγ放射源分别对其进行0.5、1、2、4、6、10 Gy照射,在照射后2、4、8、12和24 h收集各个剂量组受试者全血,提取总RNA,利用TaqMan探针实时定量PCR法,检测POLH mRNA的相对表达量。另收集30名不同年龄段(男女各占一半)的健康人外周血作为本底组,检测POLH mRNA在不同个体间的表达差异。结果:照射后2 h,仅10 Gy照射组POLH mRNA表达增加,其余各照射组无显著变化;照射后4、8、12和24 h,在0~2 Gy范围内,POLH mRNA表达量随照射剂量的增加而增加,呈现明显的剂量依赖效应,大于2 Gy照射后增加趋势接近饱和。照射后4~12 h随时间的增加表达量也不断增加,于12 h达到峰值。在对健康人群本底表达水平的分析中发现,POLH mRNA表达水平不受性别、年龄影响,在不同个体间相对均一,波动范围小。结论:POLH mRNA可由为辐射诱导表达,在一定时间和剂量范围内呈现明显的剂量依赖关系,且在不同个体之间本底表达水平相对均一,具备作为辐射损伤分子生物标志物的潜力。
OBJECTIVE: To study the changes of POLH mRNA expression in human peripheral blood exposed to γ-rays and provide experimental evidence for the establishment of POLH mRNA as a new molecular biomarker of radiation injury. Methods: Peripheral blood from 3 healthy adults was collected and irradiated with 0.5, 1, 2, 4, 6 and 10 Gy respectively with ~ (60) Coγ radioactive sources. After 2, 4, 8, 12 and 24 h The whole blood of each dose group was collected and the total RNA was extracted. The relative expression level of POLH mRNA was detected by TaqMan probe real-time quantitative PCR. Another 30 healthy people from different age groups (half male and half female) were collected as the background group to detect the difference of POLH mRNA expression in different individuals. Results: At 2 h after irradiation, the expression of POLH mRNA increased only in the group of 10 Gy irradiation, but there was no significant change in other groups. At 4, 8, 12 and 24 h after irradiation, Dose increase and increase, showing a significant dose-dependent effects, greater than 2 Gy after the increase in the trend of close to saturation. The expression increased with the increase of time from 4 h to 12 h after irradiation and peaked at 12 h. In the analysis of the background expression level of healthy people, we found that POLH mRNA expression level is not affected by gender and age, and is relatively uniform among different individuals with a small fluctuation range. CONCLUSIONS: POLH mRNA can be induced by radiation and shows a dose-dependent manner in a dose and time-dependent manner. The background expression of POLH mRNA is relatively uniform and possesses the potential of being a molecular biomarker of radiation damage.