低剂量电离辐射诱导眼晶状体上皮细胞中差异mRNA表达谱的鉴定

辐射防护 ›› 2023, Vol. 43 ›› Issue (2): 175-185.

低剂量电离辐射诱导眼晶状体上皮细胞中差异mRNA表达谱的鉴定

王萍, 范莉, 陆雪, 高玲, 刘青杰, 田梅

中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室,北京 100088

收稿日期:2022-10-25 出版日期:2023-03-20 发布日期:2023-05-11
通讯作者: 田梅。E-mail:tianmei@nirp.chinacdc.cn
作者简介:王萍(1997—),女,2019年7月毕业于山东第一医科大学应用物理学专业(医学物理学方向),现为中国疾病预防控制中心辐射防护与核安全医学所放射医学专业在读研究生。E-mail:979095051@qq.com

Identification of differential mRNA expression profiles in lens epithelial cells induced by low-dose ionizing radiation

WANG Ping, FAN Li, LU Xue, GAO Ling, LIU Qingjie, TIAN Mei

Key Laboratory of Radiological Protection and Nuclear Emergency, China CDC, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088

Received:2022-10-25 Online:2023-03-20 Published:2023-05-11

摘要/Abstract

摘要： 长期受到低剂量电离辐射的人群,眼晶状体浑浊的发生风险显著增加,阐明低剂量电离辐射诱导眼晶状体浑浊的机制,对白内障的预防和早期干预具有重要意义。本文分别以0.2 Gy ¹³⁷Cs和2 Gy ⁶⁰Co γ射线照射人晶状体上皮细胞SRA01/04,基因芯片技术筛选不同剂量照射后48 h表达量产生差异变化的mRNA,0.2 Gy与2 Gy、0.2 Gy与0 Gy、2 Gy与0 Gy比较组分别鉴定出3 690、3 304、3 970个差异表达基因。生物信息技术分析表明差异基因大多参与氧化磷酸化、细胞增殖、细胞粘附等相关通路。SRA01/04和HLE-B3细胞经不同剂量γ射线照射后24 h、48 h,用Real-time PCR验证与细胞增殖相关的基因(PIM1、HMGB1、BRE、JUN)表达变化。结果显示两种细胞中,PIM1、HMGB1和BRE基因在照射后24 h,大部分低剂量组相比于未照射组表达水平明显升高,48 h仍然有升高趋势,2 Gy照射后48 h,基因表达水平显著下降或回到基线水平。本研究表明低剂量电离辐射可以诱导人眼晶状体上皮细胞表达谱的改变,差异表达的基因参与了细胞重要的生物学过程和信号通路,有助于揭示低剂量电离辐射诱导眼晶状体浑浊并诱发白内障的分子机制。

关键词: 低剂量电离辐射, 眼晶状体浑浊, 生物信息学, 增殖

Abstract: People who exposed to low-dose ionizing radiation chronically could have an increased risk of lens opacity. It is very important to clarify the mechanism of lens opacity induced by low-dose ionizing radiation for the prevention and early intervention of cataract. This paper describes the experiments that human lens epithelial cells SRA01/04 were irradiated with 0.2 Gy ¹³⁷Cs and 2 Gy ⁶⁰Co γ-rays. Differentially expressed mRNAs were screened by gene chip technology at 48 h after irradiation, 3 690, 3 304, and 3 970 differentially expressed genes were identified in 0.2 Gy vs 2 Gy, 0.2 Gy vs 0 Gy, 2 Gy vs 0 Gy groups respectively. Bioinformatic analysis showed that the majority of these genes are associated with oxidative phosphorylation, cell proliferation and adhesion pathways. The expression of genes (PIM1, HMGB1, BRE, JUN) related to cell proliferation in SRA01/04 and HLE-B3 cells were verified by real-time PCR at 24 h and 48 h after different doses of γ-rays. The results showed that in both cell lines, the expression levels of PIM1, HMGB1, and BRE at most low-dose irradiated groups were significantly higher than those in the non-irradiated group at 24 h after irradiation, and still showed an increasing trend at 48 h after irradiation. However, at 48 h after 2 Gy irradiation, gene expression levels decreased significantly or returned to baseline. The results showed that low-dose ionizing radiation induces significant alterations of mRNAs expression in human lens epithelial cells, the differentially expressed genes play some roles in series of important biological processes and functional pathways, which could be useful to reveal molecular mechanisms of low-dose ionizing radiation induced lens opacification or cataract.

Key words: low-dose ionizing radiation, lens opacity, bioinformatics, proliferation

中图分类号:

Q345

王萍, 范莉, 陆雪, 高玲, 刘青杰, 田梅. 低剂量电离辐射诱导眼晶状体上皮细胞中差异mRNA表达谱的鉴定[J]. 辐射防护, 2023, 43(2): 175-185.

WANG Ping, FAN Li, LU Xue, GAO Ling, LIU Qingjie, TIAN Mei. Identification of differential mRNA expression profiles in lens epithelial cells induced by low-dose ionizing radiation[J]. RADIATION PROTECTION, 2023, 43(2): 175-185.

参考文献

[1] Jacobson B S. Cataracts in retired actinide-exposed radiation workers[J]. Radiat Prot Dosimetry, 2005,113(1):123-125.
[2] Vano E, Kleiman N J, Duran A, et al. Radiation cataract risk in interventional cardiology personnel[J]. Radiat Res, 2010,174(4):490-495.
[3] 张彦坤, 李玉静, 张聪瑶, 等. 低剂量辐射对X线工作者晶状体损伤效应探讨[J]. 中国医药导刊, 2016,18(09):869-870.
[4] Rastegar N, Eckart P, Mertz M. Radiation-induced cataract in astronauts and cosmonauts[J]. Graefes Arch Clin Exp Ophthalmol, 2002,240(7):543-547.
[5] Little M P, Kitahara C M, Cahoon E K, et al. Occupational radiation exposure and risk of cataract incidence in a cohort of US radiologic technologists[J]. Eur J Epidemiol, 2018,33(12):1179-1191.
[6] Su Y, Wang Y, Yoshinaga S, et al. Lens opacity prevalence among the residents in high natural background radiation area in Yangjiang, China[J]. J Radiat Res, 2021,62(1):67-72.
[7] Ahmadi M, Barnard S, Ainsbury E, et al. Early responses to low-dose ionizing radiation in cellular lens epithelial models[J]. Radiat Res, 2022,197(1):78-91.
[8] Barnard S, McCarron R, Moquet J, et al. Inverse dose-rate effect of ionising radiation on residual 53BP1 foci in the eye lens[J]. Sci Rep, 2019,9(1):10418.
[9] Worgul B V, Smilenov L, Brenner D J, et al. Mice heterozygous for the ATM gene are more sensitive to both X-ray and heavy ion exposure than are wildtypes[J]. Adv Space Res, 2005,35(2):254-259.
[10] Dalke C, Neff F, Bains S K, et al. Lifetime study in mice after acute low-dose ionizing radiation: A multifactorial study with special focus on cataract risk[J]. Radiat Environ Biophys, 2018,57(2):99-113.
[11] Bains S K, Chapman K, Bright S, et al. Effects of ionizing radiation on telomere length and telomerase activity in cultured human lens epithelium cells[J]. Int J Radiat Biol, 2019,95(1):54-63.
[12] De Stefano I, Leonardi S, Casciati A, et al. Contribution of genetic background to the radiation risk for cancer and non-cancer diseases in ptch1+/- mice[J]. Radiat Res, 2022,197(1):43-56.
[13] McCarron R A, Barnard S, Babini G, et al. Radiation-induced lens opacity and cataractogenesis: A lifetime study using mice of varying genetic backgrounds[J]. Radiat Res, 2022,197(1):57-66.
[14] Tanno B, Babini G, Leonardi S, et al. miRNA-signature of irradiated ptch1+/- mouse lens is dependent on genetic background[J]. Radiat Res, 2022,197(1):22-35.
[15] Ainsbury E A, Barnard S, Bright S, et al. Ionizing radiation induced cataracts: Recent biological and mechanistic developments and perspectives for future research[J]. Mutat Res Rev Mutat Res, 2016,770(Pt B):238-261.
[16] Stewart F A, Akleyev A V, Hauer-Jensen M, et al. ICRP statement on tissue reactions and early and late effects of radiation in normal tissues and organs—Threshold doses for tissue reactions in a radiation protection context[R].ICRP Publication 118. Ann ICRP, 2012,41(1-2):1-322.
[17] Hamada N. Ionizing radiation sensitivity of the ocular lens and its dose rate dependence[J]. Int J Radiat Biol, 2017,93(10):1024-1034.
[18] Markiewicz E, Barnard S, Haines J, et al. Nonlinear ionizing radiation-induced changes in eye lens cell proliferation, cyclin D1 expression and lens shape[J]. Open Biol, 2015,5(4):150011.
[19] Vigneux G, Pirkkanen J, Laframboise T, et al. Radiation-induced alterations in proliferation, migration, and adhesion in lens epithelial cells and implications for cataract development[J]. Bioengineering (Basel), 2022,9(1):29.
[20] Bahia S, Blais E, Murugkar S, et al. Oxidative and nitrative stress-related changes in human lens epithelial cells following exposure to X-rays[J]. Int J Radiat Biol, 2018,94(4):366-373.
[21] Wolf N, Pendergrass W, Singh N, et al. Radiation cataracts: mechanisms involved in their long delayed occurrence but then rapid progression[J]. Mol Vis, 2008,14:274-285.
[22] Zhang S, Shuai L, Wang D, et al. Pim-1 protects retinal ganglion cells by enhancing their regenerative ability following optic nerve crush[J]. Exp Neurobiol, 2020,29(3):249-272.
[23] Katakami N, Kaneto H, Hao H, et al. Role of Pim-1 in smooth muscle cell proliferation[J]. J Biol Chem, 2004,279(52):54742-54749.
[24] Mohammad G, Siddiquei M M, Alam K, et al. High-mobility group Box-1 regulates the expression of matrix metalloproteinase-9 in diabetic retina[J]. Int J Clin Exp Pathol, 2016(9):828-840.
[25] Yeung C K, Wang G, Yao Y, et al. BRE modulates granulosa cell death to affect ovarian follicle development and atresia in the mouse[J]. Cell Death Dis, 2017,8(3):e2697.