低剂量电离辐射对人淋巴细胞氧化应激及DNA损伤的影响

doi:10.3969/j.issn.1004-616x.2024.02.002

摘要/Abstract

摘要： 目的：探讨低剂量 ¹³⁷Cs γ射线照射后正常人淋巴细胞(AHH-1)是否产生氧化应激及 DNA损伤，并引发 DNA修复。方法：以剂量率为 8.32 mGy/min的 ¹³⁷Cs γ射线照射 AHH-1细胞，剂量分别为 0(未照射)、0.01、0.02、0.05、0.075、0.1和 0.2 Gy，照射后分别培养 1、24、48和 72 h。采用 CCK-8试剂盒检测细胞存活率变化；丙二醛(MDA)、超氧化物歧化酶(SOD)和活性氧(ROS)试剂盒检测细胞氧化损伤水平；免疫荧光方法分析γH2AX和53BP1焦点形成情况；实时荧光定量 PCR方法检测DNA损伤修复相关基因CDKN1A、DDB2和POLH的mRNA表达水平变化。结果：与未照射组相比，照射后24和48 h，各剂量组细胞存活率显著增强(P<0.05)；照射后48 h，MDA水平和SOD活性在0.2 Gy剂量组发生显著变化(P<0.05)；0.02~0.075 Gy和0.2 Gy剂量组ROS相对荧光强度显著升高(P<0.05)；0~0.2 Gy γ射线照后1 h，γH2AX和53BP1焦点数量随剂量增加而增加，且具有明显的剂量-效应关系(P<0.01)；与未照射组相比，照射后 48 h，DDB2和 POLH mRNA相对表达水平显著升高，差异具有统计学意义(P<0.05)。结论：低剂量电离辐射引起人淋巴细胞产生氧化应激和DNA损伤，并促进DNA损伤修复相关基因在转录水平发生改变。

关键词: 电离辐射, γ射线, 人淋巴细胞, 氧化应激, DNA损伤, 细胞增殖

Abstract: OBJECTIVE： To investigate effect of low-dose ¹³⁷Cs γ-irradiation on oxidative stress， DNA damage repair and indued DNA damage in normal human lymphocytoid cells (AHH-1). METHODS： AHH-1 cells were irradiated with ¹³⁷Cs γ-rays at a dose rate of 8.32 mGy/min，with doses of 0 (unirradiated)，0.01， 0.02，0.05，0.075，0.1，and 0.2 Gy，and cultured for 1，24，48 and 72 h after irradiation，respectively. Cell viability was examined by a CCK-8 kits. The level of oxidative damage was detected by malondialdehyde (MDA)， superoxide dismutase (SOD) and reactive oxygen species (ROS) kits. γH2AX and 53BP1 foci were performed by immunofluorescence. Changes in the expression levels of DNA damage repair-related genes CDKN1A， DDB2 and POLH mRNA were detected by real-time fluorescence quantitative PCR. RESULTS： Compared with the unirradiated group，cell viability were significantly increased at 24 and 48 h after irradiation (P<0.05). The differences in MDA levels and SOD activities were changed significantly in the 0.2 Gy (P<0.05)； the relative fluorescence intensity of ROS was significantly increased in the doses of 0.02，0.05，0.075 Gy and 0.2 Gy at 48 h after irradiation. The number of γH2AX and 53BP1 focus increased with the doses，and there were good dose-response relationships at 1 h following radiation exposure (P<0.01). Compared with the unirradiated group，the relative mRNA expression level of DDB2 and POLH genes were significantly up-regulated at 48 h after irradiation (P<0.05). CONCLUSION：Exposure to low-doses of ionizing radiation induced changes in oxidative stress，DNA damage and expression of DNA damage repair-related genes in AHH-1 cells.

Key words: ionizing radiation, gamma ray, human lymphocyte, oxidative stress, DNA damage, cell proliferation

中图分类号:

Q691.5

孙鑫, 李爽, 陆雪, 蔡恬静, 刘雅, 刘青杰, 张伟. 低剂量电离辐射对人淋巴细胞氧化应激及DNA损伤的影响[J]. 癌变·畸变·突变, 2024, 36(2): 94-99.

SUN Xin, LI Shuang, LU Xue, CAI Tianjing, LIU Ya, LIU Qingjie, ZHANG Wei. Effect of low-dose ionizing radiation on oxidative stress and DNA damage repair in human lymphocytoid cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(2): 94-99.

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