分子空物学与辐射研究

本文综述了近年来运用分子生物学的理论和技术研究放射生物学的主要进展:在DNA损伤研究方面用序列分析及限制性酶切技术对DNA链断裂及碱基损伤在分子内的定位获得了新的信息,基因探针有可能成为研究哺乳动物细胞DNA结构损伤的敏感的新方法;染色体畸变及细胞突变的分子机理已有所阐明;哺乳动物DNA修复基因的研究正在兴起阶段,已克隆出人的DNA修复基因;在哺乳动物细胞上进行外源基因转移的成就为辐射损伤的防治开辟了新的前景。     

哺乳动物细胞DNA损伤修复的研究——病毒探针的利用

用病毒探针研究DNA修复具有显著的优越性。由于病毒DNA结构及复制方式远比真核细胞的简单,病毒探针法可使真核细胞DNA修复的研究大为简化。双链DNA病毒的宿主细胞回复和多重感染回复被用来研究细胞的结构性修复功能。单链DNA病毒如细小病毒被用来探测诱导性修复过程。真核细胞内可能存在一个类似于原核细胞中的SOS功能的诱导性易错修复功能,诱导信号是DNA损伤本身。一些DNA损伤,如无碱基位损伤可充作此修复功能的底物。     

辐射诱导人肝细胞子代基因差异表达及生物信息学分析

利用基因芯片技术和实时荧光定量RT-PCR技术,采用不同剂量60Coγ射线照射人正常肝细胞系HL-7702细胞,观察比较受照细胞克隆子代基因表达图谱的变化,筛选出差异表达基因,利用生物信息学方法分析构建基因相互作用网络图,对部分差异表达基因进行验证。结果表明,与对照组相比,共同的差异表达基因有71个,其中上调的35个,...     

辐射小鼠血细胞ATM、CDKNlA、DDB2和GADD45A基因表达分析

观察小鼠全身γ射线辐照不同剂量、照射后不同时间,外周血细胞DNA损伤修复相关蛋白基因表达变化,筛选具有指示生物受照剂量潜力的指标.BALB/c小鼠,以0.8和1.6Gy/min两种剂量率,分别进行0、2、4、6和8Gy不同剂量照射.照射后4、8、12、24和48h眼眶取血,AxyPrep试剂盒抽提全血细胞总mRNA,实...     

低剂量γ射线辐照对人淋巴母细胞基因表达转录谱的影响

为探讨低剂量电离辐射生物效应作用机制,本文研究了不同剂量单次辐照后,人淋巴母细胞基因表达转录谱的变化。采用0．1Gy、0．5Gy和1．0Gy剂量的γ射线照射人淋巴母细胞,未照射细胞为对照组。照射后4h提取细胞总RNA,用含有45033个基因的人类全基因组表达谱芯片检测分析。对差异表达基因进行层次聚类分析、基因本体论分析和通路分析,并用实时荧光定量PCR验证。结果表明,3个剂量组均上调表达的基因1330个,下调表达的基因1002个。基因表达量与吸收剂量相关的基因共18个,其中与剂量正相关的基因16个,负相关的基因2个。层次聚类分析结果表明,4个实验组中,0．1Gy和1．0Gy照射组差异表达基因相似程度最高。这些差异表达的基因涉及到多条通路,如细胞周期、p53信号通路、碱基切除修复、RNA转运和内质网蛋白加工等。实时荧光定量PCR检测结果表明,CASP9（caspase9）mRNA在照射后4h随吸收剂量表达变化与基因芯片检测结果一致。基因芯片筛选出的差异表达基因有助于阐明低剂量辐射生物效应作用机理,与辐射剂量相关的差异表达基因有可能成为低剂量辐射暴露的生物剂量计。     

居民由食物所致内剂量估算方法

环境放射性包括天然和人工放射性核素两类。食品是环境放射性进入人体主要途径之一,涉及的人群是比放射性职业人员大得多的广大居民。监测食品环境放射性污染对居民危害过去是依据关键器官的剂量当量率,按ICRP新基本建议书,则是依据有效剂量     

微细胞介导染色体转移法提高AT5 BIVA细胞对电离辐射的抗性

ＡＴ５ＢＩＶＡ细胞是一株经ＳＶ４０病毒转化的ＡＴ病人皮肤成纤维细胞，对γ射线高度敏感。实验用ＦＤ３（含人第１１号染色体的人鼠杂种细胞）、ＦＤ８（不含人第１１号染色体的人鼠杂种细胞）、ＬＭ／ＴＫ鼠细胞）为洪体，通过微细胞介导染色体转移（ＭＭＣＴ）向ＨＴ５ＢＩＶＡ细胞导入人或鼠的完整染色体，经两次３Ｇｙγ射线照射筛选后，获得ＡＴ５ＢＩＶＡ与ＦＤ３微细胞融合的杂合细胞ＡＴ／ＦＤ３－１，对γ射线抗性有显著提高。而ＦＤ８或ＬＭ／ＴＫ的微细胞与ＡＴ５ＢＩＶＡ细胞的杂合细胞，对γ射线抗性未增加。枝型分析表明ＡＴ／ＦＤ３—１细胞中包含了来源于ＦＤ３细胞的人第１１，１４号染色体和数条鼠染色体。通过对照实验，排除了人１４号和鼠染色体提高ＡＴ／ＦＤ３－１细胞对γ射线抗性的可能性．确认人第１１号染色体与ＡＴ细胞对电离辐射敏感性相关，提示人第１１号染色体上可能存在决定细胞对γ射线抗性的相关基因。     

Genetic susceptibility to cancer: ICRP Publication 79

A Task Group of the ICRP Committee 1 (Radiation Effects) has reviewed relevant data with the objective of advising the Main Commission of the ICRP on the possible implications for radiological protection of emerging views on genetic susceptibility to cancer (Chapter 1).Chapter 2 considers DNA damage and its processing/repair after ionising radiation and serves principally to demonstrate that a few rare cancer-prone, human recessive genetic disorders show DNA repair deficiency and profound increases in radiosensitivity. Less dramatic changes in radiosensitivity are also apparent in a wider range of such disorders. The cellular mechanisms that underly the association between DNA damage processing and tumorigenesis are discussed.Chapter 3 reviews the mechanisms and genetics of solid tumours illustrating the ways in which mutations in proto-oncogenes, tumour suppressor genes together with those in DNA repair and cell cycle control genes can contribute to tumour development. Specific examples are given of how germ line mutation of such genes can predispose to familial cancer. It is judged that up to 5% of all solid tumours have a recognisable genetic component. Heritable organ-specific effects are most usual and cancers of the breast and colon tend to show the most obvious genetic components. Clearly discernible genetic effects are seen when rare dominant germ line mutations express strongly as familial cancer (high penetrance mutations), but the existence of perhaps less rare low penetrance mutations and gene–gene interactions are recognised but not well understood.Chapter 4 considers the mechanisms and genetics of lympho-haemopoietic tumours. Specific chromosomal translocations and proto-oncogene activation events are much more frequent in human leukaemia/lymphoma than in solid tumours. Genetic predisposition to leukaemia/lymphoma is found in a number of non-familial recessive genetic disorders of DNA processing and/or chromosomal instability. Familial manifestation of susceptibility to these tumours is, however, extremely rare. The genetic component, although poorly defined, is judged to be less than that of solid tumours and expressed largely in childhood.Chapter 5 reviews and discusses limited data that comment upon tumorigenic radiosensitivity in cancer-prone genetic conditions. From knowledge of the fundamental processes involved it is judged that in most, but not all, cases genetic susceptibility to spontaneous tumours will be accompanied by a greater-than-normal risk after radiation. A review of epidemiological, clinical and experimental data relevant to this issue suggests that although a wide range of different sensitivities may be involved, a factor of 10 increase in sensitivity broadly accords with the limited human data available. This interim judgement of a factor of 10 increase in radiation risk in such human genetic disorders is made for the purposes of illustrative modelling and calculation. In addition, specific attention is given to breast cancer risk in heterozygotes for the radiosensitive human disorder, ataxia-telangiectasia; this association, while in no way discounted, is judged to be less strong than that claimed by some.Chapter 6 discusses and develops computational modelling procedures that aim to describe the impact of genetic factors on radiation-tumorigenesis in human populations. Estimates of the prevalence of known cancer-prone genetic disorders are made but breast cancer susceptibility is used to illustrate the application of the model developed. The most important message to emerge from this work is that, even at an assumed high level of radiation sensitivity, the prevalence of familial (high penetrance) genetic disorders in the population is too low (<1%) for there to be a significant impact on risk in typical human populations. In principle, however, there is the potential for such impact in atypical inbred sub-populations where these mutations can be more common. These modelling procedures are also used to illustrate how incomplete penetrance of these mutations will dilute any impact on population risk.In conjunction with the Main Commission of the ICRP, in Chapter 7 the Task Group discusses the potential implications of the main report for radiological protection. Their principal conclusions are: (i) That current estimates of radiation cancer risk already include an unknown contribution from genetically radiosensitive sub-populations. (ii) Using the data cited, the likely contribution to radiation risk from familial cancer disorders is too low to generate an unacceptable distortion of current estimates of cancer risk in the vast majority of human populations. (iii) There is insufficient knowledge to judge the contribution to risk from mutations of low penetrance that do not express as familial cancer. (iv) Because of the high risk of spontaneous cancer in familial disorders, low doses of radiation (say 100 mSv) are most unlikely to impact significantly on life-time cancer risk in an affected individual; at high doses, such as those experienced in radiotherapy, this relative risk may however become important. (v) Because organ-specific cancer risk is predicted in most familial disorders, the absolute increase overall in risk to an affected individual will be diluted, ie. comparing normal and affected individuals. (vi) The utility of genetic testing for cancer predisposition in the context of radiological protection is currently limited by technical factors and concerns on predictive power. In the future genetic testing may find selected use prior to certain medical exposures to radiation, but the value of such procedures, as applied to low-dose occupationally exposed individuals, is open to doubt; it would also be subject to major ethical scrutiny outside the remit of the ICRP.The Task Group and the Main Commission of the ICRP stress that, because of the current lack of knowledge, the above judgements should be regarded as preliminary. The report serves principally to provide a framework on which to develop further views in this rapidly advancing area of human genetics.     

以斑马鱼为模式生物的辐射效应修饰剂研究

利用斑马鱼作为模式生物,进行一系列的实验研究,尤其在胚胎发育、基因表达、药物筛选和疾病模型方面有其独特的优势。但它被应用于辐射效应修饰剂的研究时间较短,已有的文献显示发展前景良好。本工作就近些年以斑马鱼为模式生物研究的辐射效应修饰剂进行概述,主要涉及如氨磷汀、DF-1、AG1478、Flavopiridol和一些DNA损伤修复蛋白。     

Tip60过表达对细胞DNA辐射损伤修复及细胞周期的影响

为了探讨Tip60对细胞DNA损伤修复及细胞周期的影响及其相关机制,通过在U2OS细胞中稳定转染外源Tip60基因,分析了细胞增殖能力及DNA双链断裂修复能力,以及辐射后引起的细胞周期阻滞和细胞周期相关蛋白表达变化。结果发现Tip60在U2OS细胞中的稳定表达降低了细胞增殖能力却提高了细胞DNA损伤修复能力,并通过引起电离辐射诱发CyclinB/CDC2复合物表达水平下降,导致细胞G2/M期阻滞延长。