Survivin表达在高LET射线诱导细胞凋亡中

先前的研究表明, 肿瘤细胞中survivin的高表达与细胞对高传能线密度(LET)射线的辐射抗性相关。 研究了survivin表达在高LET射线诱导的细胞凋亡中的作用, 发现抑制survivin表达对高LET C离子辐射诱导的Bcl-2和Bax表达没有明显的影响。 在高LET射线辐照中, survivin可能通过抑制caspase-3和-9活性的途径, 抑制了细胞凋亡。It has been proven that over expression of survivin in cancerous cell lines is related to the radioresistance of cells to high LET radiation in previous work. In this study, action mechanisms of survivin gene in apoptosis induced by high LET radiation were investigated. We found that inhibiting survivin by siRNA had no notable influence on Bcl-2 and Bax expressions induced by carbon ions. Survivin depressed cell apoptosis through the inhibition of the activities of caspase-3 and -9 possibly in cell apoptosis induced by high LET radiation.     

辐射诱导的DNA修复与细胞凋亡的ATP调控

综述了DNA辐射损伤导致的细胞阻遏于G1期以及在该时期对DNA的修复活动, 提出了较大剂量辐射诱导的三磷酸腺苷不足导致细胞凋亡的假说, 并分析了细胞走向凋亡与修复的辨正关系。 DNA damage induced by irradiation,which makes the cell arrested at G1 stage and DNA repair being activated in this stage,are summarized. It is proposed that the deficiency of adenosine triphosphate which is induced by the larger irradiation dose, induces cell apoptosis. And the relationship of cell selecting repair and apoptosis is also analyzed.     

和厚朴酚对非小细胞肺癌细胞的辐射增敏效应

研究了和厚朴酚（HNK）对非小细胞肺癌（NSCLC）细胞系A549和H1299对低线性能量转移（LET） X射线和高LET碳离子的辐射增敏效应。首先用CCK-8检测了HNK对A549和H1299细胞的生长抑制情况,发现20 μmol/L的HNK处理对细胞的生长抑制作用较弱。用该浓度HNK预处理细胞2 h后给予不同剂量X射线或碳离子的照射,克隆存活法检测细胞的辐射敏感性,Annexin-PI双染法检测细胞凋亡,γH2AX焦点法检测DNA的双链断裂（DSB）损伤。实验结果显示:与X射线相比,NSCLC细胞对碳离子更敏感,HNK预处理仅对碳离子照射有辐射增敏作用;与碳离子单独照射相比,HNK预处理联合碳离子照射诱导了更明显的细胞凋亡;在照射后24 h,HNK预处理联合碳离子照射引起的细胞γH2AX焦点阳性率维持在较高水平,而X射线照射没有这些效应。实验结果表明,HNK预处理抑制了NSCLC细胞DNA的DSB修复,诱导了细胞凋亡的发生,从而提高了细胞对碳离子的辐射敏感性。The radiosensitizing effect of Honokiol (HNK) on non-small cell lung carcinoma (NSCLC) cell lines A549 and H1299 to low-linear energy transfer (LET) X-rays and high-LET carbon ions was investigated in this study. First, the inhibitory effects of HNK on the growth of A549 and H1299 cells were detected by CCK-8 assay, and 20 μmol/L HNK treatment was found to induce a growth inhibitory effect slightly in these two cell lines. Cells were pre-treated with HNK and then irradiated with X-rays and carbon ions of different doses. Cellular radiosensitivity, apoptosis and DNA damage were analyzed by clonogenic survival, Annexin-PI staining and γH2AX foci, respectively. The results showed the cells were more sensitive to carbon ion irradiation compared to X-rays and the radiosensitization of HNK was only observed after carbon ion irradiation. Furthermore, the co-treatment led to higher apoptosis rate 48 h after irradiation and increased the positive rate of γH2AX foci 24 h after irradiation in A549 and H1299 cells compared with those in the groups treated with carbon ion irradiation alone. These phenomena were not observed after X-ray irradiation. Our data suggest that the pre-treatment with HNK inhibited DNA DSB repair, induced apoptosis and then enhanced the cellular radiosensitivity to carbon ions in NSCLC cells.     

电离辐射引起的线粒体DNA损伤及突变研究进展

指出了线粒体DNA损伤及突变研究与核DNA相关研究的区别,并总结了国内外关于电离辐射引起线粒体DNA损伤突变的研究进展。 针对放射生物学中线粒体学方面研究还有待进一步加强的现状, 中国科学院近代物理研究所的研究人员在方法学上建立起了real-time PCR和long PCR等手段对线粒体DNA损伤及突变进行定量检测, 并以此为基础, 对电离辐射引起的线粒体功能变化进行了较为深入的研究。总结了线粒体DNA损伤及突变研究对阐明电离辐射引起的生物学效应所具有的贡献, 提出未来在放射生物学中以远后效应, 能量代谢为主的线粒体学研究方向。 Current advance in ionizing radiation induced mitochondrial DNA damage and mutations is reviewed, in addition with the essential differences between mtDNA and nDNA damage and mutations. To extent the knowledge about radiation induced mitochondrial alterations, the researchers in Institute of Modern Physics, Chinese Academy of Sciences developed some technics such as real time PCR, long PCR for accurate quantification of radiation induced damage and mutations, and in depth investigation about the functional changes of mitochondria based on mtDNA damage and mutations were also carried out. In conclusion, the important role of mitochondrial study in radiation biology is underlined, and further study on mitochondrial study associated with late effect and metabolism changes in radiation biology is pointed out.     

特异性抗癌因子——凋亡素

凋亡素是一种抗癌因子, 能以非p53依赖性途径诱导不同类型人肿瘤细胞的凋亡, 不受Bcl 2的抑制作用, 且对正常细胞不起作用。 根据凋亡素的肿瘤特异性, 可将其作为一种极具潜力的抗肿瘤生物制剂, 在人体内大量传递给肿瘤细胞, 有选择性地根除肿瘤细胞。 论述了凋亡素抗肿瘤细胞的作用机理, 并对其应用现状、 前景及进一步研究的主要问题予以简要评述。Apoptin is an anti cancer gene. It can induce p53 independent,Bcl 2 insensitive type of apoptosis in various human tumor cells,and fails to induce programmed cell death in normal cell. Because of having tumor specificity, apoptin can be transferred enough to tumor cell in vivo,selectly kill the tumor as a potential anti tumor biological preparation. In this paper, we provide a brief review on the anti cancer mechanism of apoptin, current status and application prospect, and the main issues in apoptin studies.     

Study on Plasmid and Damage Induced by Low-energy Neon Ion Irradiation

DNA is considered to be the most important and sensitive target in biological systems. Beside base damage, DNA strand breaks are the major lesion in the genome due to exposure to ionizing radiation. Mutation can be introduced to DNA as a result of enzymatic processing of DNA lesions or post-irradiation replication. However, the mechanisms of radiation-induced mutations are not well clarified at the molecular level. A good way to approach the mechanism is to irradiate the plasmid DNA of heavy ion, then transfect the DNA to host cells to determine the mutation spectra. So to study the effect of heavy ions on the simple plasmid DNA is even predominant or more feasible.     

辐射致DNA损伤及相关生物-化学-物理问题(英文)

介绍了在DNA辐射损伤以及相关生物、物理、化学问题的一些研究工作。重点是径迹结构、DNA双链断裂和细胞学终点的关系。P53-Mdm2负反馈回路在DNA损伤的细胞响应方面起重要作用,用一个简单的模型研究了P53-Mdm2负反馈回路相互作用的动力学行为。Some of recent works are presented on radiation induced DNA damage being carried out at China Institute of Atomic Energy （CIAE） and related bio-chemi-physical problems. Emphasis is placed on track structure and its relation to the cell end-point, and a simple model proposed to study the dynamical behavior of P53-Mdm2 interaction which plays pivotal role in cellular response to DNA damage.     

重离子对人胃癌细胞DNA错配修复基因MSH2 表达的影响

采用高传能线密度(LET) 重离子辐照人胃癌SGC7901 细胞,应用流式细胞技术、蛋白质印迹法(Western blot) 及反转录聚合酶链式反应(RT-PCR) 观察重离子诱导人胃癌SGC7901 细胞周期、凋亡和MSH2 表达状况。结果表明: 与对照组相比,SGC7901 细胞在辐射后72 h G2/M 期所占细胞比率(33.26±0.08) 和凋亡率(24.16±0.64) 均达到峰值,且呈时间依赖性增加;经重离子照射后,DNA错配修复基因MSH2 mRNA 和蛋白表达水平在6 h 最高。结果提示:重离子在体外诱导SGC7901 细胞周期阻滞和凋亡,且具有显著的时间依赖性效应;重离子在一定剂量和时间下,诱导了SGC7901 细胞MSH2 基因表达。DNA错配修复基因MSH2 可能参与了重离子辐照诱导胃癌细胞DNA损伤的修复应答。Human gastric cancer cell SGC7901 were irradiated with high linear energy transfer (LET) carbon ion. Apoptotic cells after irradiation were analyzed by flow cytometry and expression of MSH2 genes in the irradiated cells was detected by western blot and RT-PCR assay. Compared with the control group, we found that the number of G2/M (33.26±0.08) or apoptosis (24.16±0.64) of SGC7901 cells reached a maximum after irradiation at 72 h in a dose dependent manner. And heavy ion irradiation efficiently up-regulated the expression of MSH2 gene at 4.0 Gy after being irradiated 6 h. These results imply that heavy ion beam could induce cell apoptosis and cell cycle arrest in time-dependent manners. Furthermore, expression of MSH2 genes activated by carbon ion irradiation suggests that DNA mismatch repair gene MSH2 might be involved in DNA repair pathways.     

Automated damage diagnostic system for laser damage threshold tests

<正>An automated damage diagnostic system for collecting plasma flash is developed to diagnose damage in a laser-induced damage threshold(LIDT) test system.Experiment is done to verify the accuracy of this system and analyze the relationship between the plasma signals and the damage morphologies.The results obtained by the system are found to be in excellent agreement with those obtained by the much laborious method of Normaski microscope.Results show that plasma signals above 1 V correspond to the damage morphology of surface discolorations with or without pits in their centers,and plasma signals below or just around 1 V correspond to the damage morphology of pits.The misdiagnosis is attributed to contaminations and air breakdown.     

辐射诱发体内旁效应特征的研究进展

大量的细胞生物学研究显示, 辐射诱发旁效应的传输信号包括活性氧自由基（ROS）、 一氧化氮自由基（NO）以及一些细胞因子。 近年来,越来越多的文献报道关于体内旁效应特征的研究, 并已经证实在生物体内辐射诱导的旁效应不仅能发生在相邻组织, 还可发生在远源器官。这些体内旁效应包括DNA损伤、表观遗传学改变、miRNA及基因表达改变、 细胞增殖和凋亡等。 为了总结和分析辐射诱发体内旁效应的特征, 本文综述了辐射诱发体内旁效应的特点, 包括其与性别、 传播途径、辐射品质的关系以及相关机制的研究。 Radiation induced bystander effect is defined as the induction of damage in neighboring non hit cells by signals released from directly irradiated cells. ROS, NO and cytokines are involved in signaling pathways of bystander effects. Recently, the bystander effects in vivo have been reported more and more. It has been indicated that radiation induced bystander effect was localized not only in bystander tissues but also in distant organs. This effect includes many biological endpoints such as DNA damage, epigenetic, miRNA and gene expression changes, cell proliferation and apoptosis. In this review we described different aspects of ionizing radiation induced bystander effects such as its characteristics, sex specific, signaling transfer, dose and LET dependence, and related mechanisms.     

基于小波变换和图论方法识别乳腺微钙化点簇

成簇的微钙化点是早期乳腺癌的一个重要表征,用计算机辅助诊断时需要在乳腺片图像中判断有无钙化点簇。提出了一种利用Daubechies小波对乳腺图像进行增强,用二维熵阈值分割法对增强后的图像进行阈值分割,然后采用图论方法提取出成簇钙化点的方法。从而实现了计算机辅助诊断系统辅助医师判断恶性钙化。     

不同LET辐射致DNA集簇性损伤的研究

利用辐照质粒DNA构象变化的分子模型,以DNA糖苷酶Fpg和AP核酸内切酶EndoIII识别并切割辐射所致DNA碱基损伤,将其转换为DNA断裂损伤,通过电泳分析DNA分子构象变化,研究比较γ射线、质子和7Li离子诱发DNA集簇性损伤。50Gy以上高剂量γ辐射对质粒DNA的损伤主要表现为单链断裂(SSB)和很少比例的双链断裂(DSB),并能产生一定水平的集簇性损伤。相比之下,高能质子束和高LET的7Li离子直接所致DNA的断裂损伤以及所产生的集簇性碱基损伤比γ射线的要严重,质子10Gy照射就可诱发明显的集簇损伤。     

重离子诱导细胞衰老研究

综述了细胞衰老诱导因素和衰老细胞的特征,提出细胞衰老在癌症发生发展过程中具有双重作用效应。同时,以X射线和12C6+离子束对黑色素瘤细胞系92-1或人正常成纤维细胞系MRC5进行辐照,观察分析DNA损伤应答效应和细胞衰老诱导。实验结果表明,相对于X射线,12C6+离子束能导致DNA团簇损伤,持续激活DNA损伤应答,更容易诱导细胞衰老;12C6+离子束能同时诱导癌细胞和正常细胞衰老。这些实验结果暗示开展12C6+离子束诱导细胞衰老研究具有紧迫性。     

Mdm2生成速率调控的p53-Mdm2振子的全局动力学和稳定性

肿瘤抑制蛋白p53的动力学在一定程度上可以决定DNA损伤后的细胞命运.p53的动力学行为与p53信号通路中p53-Mdm2振子模块密切相关.然而,p53的负调控子Mdm2的生成速率的增加使其在一些癌细胞中过表达.因此探讨Mdm2生成速率对p53动力学的影响有重要意义.同时,PDCD5作为p53的激活子也调控p53的表达.因此,本文针对PDCD5调控的p53-Mdm2振子模型,通过分岔分析获得了Mdm2生成速率所调控的p53的单稳态、振荡以及单稳态与振荡共存的动力学行为,且稳定性通过能量面进行了分析.此外,噪声强度对p53动力学的稳定性有重要的影响.因此,针对p53的振荡行为,探讨了噪声强度对势垒高度和周期的影响.本文所获得的结果对理解DNA损伤后的p53信号通路调控起到一定的指导作用.     

Multiscale approach to radiation damage induced by ion beams: complex DNA damage and effects of thermal spikes

We present the latest advances of the multiscale approach to radiation damage caused by irradiation of a tissue with energetic ions and report the calculations of complex DNA damage and the effects of thermal spikes on biomolecules. The multiscale approach aims to quantify the most important physical, chemical, and biological phenomena taking place during and following irradiation with ions and provide a better means for clinically-necessary calculations with adequate accuracy. We suggest a way of quantifying the complex clustered damage, one of the most important features of the radiation damage caused by ions. This quantification allows the studying of how the clusterization of DNA lesions affects the lethality of damage. We discuss the first results of molecular dynamics simulations of ubiquitin in the environment of thermal spikes, predicted to occur in tissue for a short time after an ion’s passage in the vicinity of the ions’ tracks.     

A clustered speckle approach to optical trapping

An in situ study of the clustered speckle 3D structure using an optical tweezer setup is presented. Clustered speckles appear when a coherently illuminated diffuser is imaged through a pupil mask with several apertures, properly distributed over a closed path, which is placed before the objective lens of a standard optical trapping system. Thus, light volumes are reduced several times when compared with standard speckles, being even smaller than the focus volume of a Gaussian beam commonly used to trap. Moreover, clustered speckles have odd statistical properties which differentiated it from standard speckles. Then, geometrically ordered multiple trapping arrays, with statistical random distribution of intensities, can be created with this technique. This fact could enable different studies concerning optical binding or new developments in coherent matter wave transport where Optical Trapping has been proven with standard speckles. In this work, a qualitative analysis of clustered speckles in an optical tweezer setup relative to the number of apertures in the mask and their size is carried on. Besides, in the Rayleigh regime, a general quantitative method to characterize the trapping capability of an optical field is proposed. Then, it is applied to clustered speckles. As a result, a relation between aperture size and the maximum size of the particles that could be trapped is found. This fact opens the possibility of engineering the statistic of the trapped particles by properly selecting the pupil mask.     

Plasmid DNA damage by heavy ions at spread-out Bragg peak energies

Interaction of ionizing radiation with plasmid DNA can lead to formation of single strand breaks, double strand breaks and clustered lesions. We have investigated the response of the synthetic plasmid pBR322 in aqueous solution upon irradiation with ¹²C ions under spread-out Bragg peak conditions (densely ionizing) and with ¹³⁷Cs γ-photons (sparsely ionizing) as a function of dose. To evaluate the relevance of indirect effects, i.e. influences of diffusion limited radical induced DNA damage triggered by water radiolysis, the experiments were performed at various concentrations of the radical scavenger mannitol. Agarose gel electrophoresis was employed to quantify the DNA damage. At low scavenger concentration for a given dose DNA damage is higher for γ-photons than for ¹²C. For the latter, the microscopic dose distribution is inhomogeneous, with very high dose deposited along the few tracks through the solution. This is in agreement with the concept that scavengers efficiently reduce damage for γ-photons, implying that the underlying damage mechanism is single strand break induction by OH radicals. For ¹²C induced damage, the fraction of SSB and DSB that is unaffected by radical scavengers and thus due to direct effect is quantified.     

Molecular devices for high fidelity of DNA replication and gene expression

Certain types of DNA lesions, produced through cellular metabolic processes and also by external environmental stresses, are responsible for the induction of mutations as well as of cancer. Most of these lesions can be eliminated by DNA repair enzymes, and cells carrying the remaining DNA lesions are subjected to apoptosis. The persistence of damaged bases in RNA can cause errors in gene expression, and the cells appear to possess a mechanism which can prevent damaged RNA molecules from entering the translation process. We have investigated these processes for high fidelity of DNA replication and gene expression, by using both biochemical and genetic means. We herein describe (1) the molecular mechanisms for accurate DNA synthesis, (2) mammalian proteins for sanitizing the DNA precursor pool, (3) error avoidance mechanisms for gene expression under oxidative stress, and (4) the roles of DNA repair and apoptosis in the prevention of cancer.     

Radiation damage to DNA: the importance of track structure

A wide variety of biological effects are induced by ionizing radiation, from cell death to mutations and carcinogenesis. The biological effectiveness is found to vary not only with the absorbed dose but also with the type of radiation and its energy, i.e., with the nature of radiation tracks. An overview is presented of some of the biological experiments using different qualities of radiation, which when compared with Monte Carlo track structure studies, have highlighted the importance of the localized spatial properties of stochastic energy deposition on the nanometer scale at or near DNA. The track structure leads to clustering of damage which may include DNA breaks, base damage etc., the complexity of the cluster and therefore its biological repairability varying with radiation type.

The ability of individual tracks to produce clustered damage, and the subsequent biological response are important in the assessment of the risk associated with low-level human exposure. Recent experiments have also shown that biological response to radiation is not always restricted to the ‘hit’ cell but can sometimes be induced in ‘un-hit’ cells near by.