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目的探讨电离辐射诱导人正常肝细胞HL-7702甲型肝炎病毒受体2(HAVCR2)和MXR7基因的表达变化,为寻找辐射致肝脏损伤早期诊断标志物提供实验依据。方法将对数生长期的人正常肝细胞HL-7702分为10组,利用中国辐射防护研究院钴-60(60Co)照射装置对各组肝细胞照射,吸收剂量率为1.29 Gy/min,吸收剂量分别为0、0.2、0.4、0.6、0.8、1.0、2.0、3.0、4.0、5.0 Gy,利用实时荧光定量聚合酶链反应(PCR)技术检测肝细胞HL-7702 HAVCR2和MXR7基因的变化,分析剂量-反应关系。结果 HAVCR2基因表达在照射后4、24 h总体上调;其中,0.2～0.8 Gy剂量组在照射后24 h基因相对表达量与照射剂量具有很好的剂量-反应关系,满足回归方程:y=0.14x2-0.48x+1.34;1.0～3.0Gy剂量组在照射后24 h,基因相对表达量与照射剂量呈现很好的剂量-反应关系,满足回归方程:y=0.44 x+0.56。MXR7基因表达在照射后4、24 h较对照组显示出明显差异;其中,0.2～0.8 Gy剂量组MXR7基因表达在照射后4 h与对照组相比总体上调,但随照射剂量增大,上调幅度降低;照射后24 h,其基因表达未见明显规律;1.0～5.0 Gy剂量组MXR7基因表达在照射后24 h整体上调,且基因的表达与照射剂量呈现一定的剂量相关性,满足方程为y=0.57x+0.43。结论电离辐射可以诱导人正常肝细胞HL-7702 HAVCR2和MXR7基因的差异表达,且基因表达与照射剂量存在一定的剂量相关性。
Objective To investigate the expression changes of hepatitis A virus receptor 2 (HAVCR2) and MXR7 gene in human normal hepatocytes HL-7702 induced by ionizing radiation and provide experimental evidence for finding early diagnostic markers of liver damage caused by radiation. Methods Human normal hepatocytes HL-7702 in logarithmic growth phase were divided into 10 groups. The irradiation dose was 1.29 Gy / min for each group of hepatocytes irradiated by cobalt-60 (60Co) irradiation device of China Institute of Radiation Protection. Absorption The dosages were 0,0.2,0.4,0.6,0.8,1.0,2.0,3.0,4.0,5.0 Gy respectively. The changes of HAVCR2 and MXR7 gene in HL-7702 hepatocytes were detected by real-time fluorescence quantitative polymerase chain reaction (PCR) Dose-response relationship. Results The expression of HAVCR2 gene was up-regulated 4 and 24 h after irradiation. Among them, the relative expression level of gene at 24 h after irradiation in 0.2 ~ 0.8 Gy group had a good dose-response relationship with the irradiation dose, which satisfied the regression equation: y = 0.14 x2-0.48x + 1.34. At 24 h after irradiation, the relative gene expression of 1.0-3.0Gy dose-dependently dose-response relationship showed that the regression equation was: y = 0.44 x +0.56. MXR7 gene expression at 4 and 24 h after irradiation showed a significant difference compared with the control group; the expression of MXR7 gene in 0.2-0.8 Gy dose group was generally up-regulated compared with the control group at 4 h after irradiation, but was up-regulated as the radiation dose was increased There was no obvious regulation of gene expression at 24 h after irradiation. The expression of MXR7 gene in 1.0-5.0 Gy group was up-regulated at 24 h after irradiation, and the expression of the gene was dose-dependent with the irradiation dose, y = 0.57x + 0.43. Conclusion Ionizing radiation can induce the differential expression of HAVCR2 and MXR7 genes in human normal hepatocytes HL-7702, and there is dose-dependent relationship between gene expression and irradiation dose.