辐射致DNA和细胞损伤的几种检测技术初探
|
摘要
随着质子和重离子治疗肿瘤的应用日益为人们所接受,高传能线密度(LET)辐射哺乳动物细胞的生物学效应机理研究越来越受到重视。
本研究利用琼脂糖凝胶电泳配合先进的成像系统技术、集落形成实验等传统的细胞生物学分析手段以及基因芯片这一现代分子生物学实验技术研究了辐射致质粒DNA和一些细胞的生物学效应,以期为重离子和质子治癌、硼中子俘获治疗(BNCT)、太空辐射和核辐射的危险性评估及其防护等提供有价值的实验依据。实验过程及初步结果如下:
利用60Co-γ射线对不含和含自由基清除剂的水溶液pUC19质粒DNA进行辐照,使用凝胶电泳方法分析实验样品,结果表明在水状DNA溶液环境下,γ射线辐射过程中产生的自由基是诱发DNA单链和双链断裂的重要原因;甘露醇,维生素C和茶多酚是很好的自由基清除剂,在相同条件下甘露醇比维生素C具有更强的防护作用;对比以前做过的重离子辐照实验,发现自由基清除剂对γ射线产生自由基的清除作用更加明显,这对于评估重离子辐照中自由基的作用具有十分重要的参考价值。
利用60Co-γ射线和238Puα源发射的α粒子对U251神经胶质瘤细胞和HepG2肝癌细胞进行辐照,结果表明:U251细胞比HepG2细胞具有更高的辐射敏感性;α粒子比γ射线对细胞增殖的抑制作用更加明显;在同等剂量下,α粒子与γ射线相比具有更高的相对生物学效应(RBE)。在兰州近代物理研究所重离子研究装置(HIRFL)辐照终端,用5Gy能量为80MeV/u的12C6+离子对L02正常肝细胞进行了辐照,用22K的人类全基因组寡核苷酸芯片分析基因转录谱,结果表明辐照后培养6小时细胞发生差异表达的基因有37个,其中19个基因表达下调,18个基因表达上调。辐照后培养24小时细胞发生差异表达的基因有269个,202个基因表达下调,67个基因表达上调。这些基因大多与细胞信号转导、细胞周期调控、DNA损伤修复、细胞凋亡等相关。实验结果为阐明重离子辐射效应的分子机制提供了有用信息。
在进行上述实验研究的同时,学习和掌握了辐射致DNA和细胞损伤的几种检测技术,并对它们的适用性和优缺点进行了评估。
Proton and heavy ions have been applied in cancer radiation therapy field, and people have accepted this method increasingly. Under this background, the research on the biological effects and mechanisms of mammalian cell irradiated by high linear energy transfer (LET) ions has become more and more important.
This research investigate the biological effects of plasmid DNA and several cells after irradiation by methods of agarose gel electrophoresis along with advanced gel imaging system, colony forming ability experiment along with other traditional cell biology techniques and gene chip as a modern molecular biotechnology. The experimental results of this research can be expected to offer valuable basic data for cancer therapy by heavy ions and protons radiation method, Boron Neutron Capture Therapy (BNCT) as well as the dangerous assessment and protection of human in space or nuclear radiation environment, etc. The experimental procedure and elementary results are as follows:
The pUC19 plasmid DNA with or without free radical scavengers in aqueous solution is irradiated by 60Co-γrays, the results are analyzed by gel electrophoresis and Alpha Innotech digital imaging system .The results indicate that free radicals produced byγrays radiation are an important factor for the induction of DNA single strand breaks (SSB) and DNA double strand breaks (DSB). Mannitol, Vitamin C and tea polyphenols are all good scavengers, and mannitol is more effective than Vitamin C under the same condition. Compared with heavy ions, the capacity of scavengers is more remarkable on free radicals induced byγrays, which is more important to evaluate the effect of free radicals in heavy ion radiation.
U251 glioma cell and HepG2 liver cancer cell are irradiated by 60Co-γrays andαparticles emitted from 238Puαsource respectively. The results indicate that U251 cell is more sensitive to ionizing radiation than HepG2 cell, andαparticles irradiation inhibit the cell proliferation more effectively thanγrays irradiation. It means thatαparticles have a higher relative biological effectiveness (RBE) thanγrays.
L02 normal liver cell is irradiated by 5Gy 80MeV/u 12C6+ ions in radiation terminal of Heavy Ion Research Facility at Lanzhou (HIRFL). 22k human genome oligonucleotide microarray was used to explore the transcriptional profiles of the cells at 6h and 24h post-irradiation. In the 6h post-irradiation cells, the microarrays displayed 37 differentially expressed genes, including 18 up-regulated genes and 19 down-regulated genes. In the 24h post-irradiation cells, the microarrays displayed 269 differentially expressed genes, including 67 up-regulated gene and 202 down-regulated genes. Most of these changed genes were associated with signal transduction,cell cycle control,DNA repair and apoptosis. These observations provided the information of radiation-induced genes for elucidating molecular mechanisms of the diverse biological effects generated by heavy ion irradiation.
During the experiment mentioned above, several testing techniques on DNA and cell damage have been learnt and the adaptability,advantages and disadvantages of these methods are evaluated.
本研究利用琼脂糖凝胶电泳配合先进的成像系统技术、集落形成实验等传统的细胞生物学分析手段以及基因芯片这一现代分子生物学实验技术研究了辐射致质粒DNA和一些细胞的生物学效应,以期为重离子和质子治癌、硼中子俘获治疗(BNCT)、太空辐射和核辐射的危险性评估及其防护等提供有价值的实验依据。实验过程及初步结果如下:
利用60Co-γ射线对不含和含自由基清除剂的水溶液pUC19质粒DNA进行辐照,使用凝胶电泳方法分析实验样品,结果表明在水状DNA溶液环境下,γ射线辐射过程中产生的自由基是诱发DNA单链和双链断裂的重要原因;甘露醇,维生素C和茶多酚是很好的自由基清除剂,在相同条件下甘露醇比维生素C具有更强的防护作用;对比以前做过的重离子辐照实验,发现自由基清除剂对γ射线产生自由基的清除作用更加明显,这对于评估重离子辐照中自由基的作用具有十分重要的参考价值。
利用60Co-γ射线和238Puα源发射的α粒子对U251神经胶质瘤细胞和HepG2肝癌细胞进行辐照,结果表明:U251细胞比HepG2细胞具有更高的辐射敏感性;α粒子比γ射线对细胞增殖的抑制作用更加明显;在同等剂量下,α粒子与γ射线相比具有更高的相对生物学效应(RBE)。在兰州近代物理研究所重离子研究装置(HIRFL)辐照终端,用5Gy能量为80MeV/u的12C6+离子对L02正常肝细胞进行了辐照,用22K的人类全基因组寡核苷酸芯片分析基因转录谱,结果表明辐照后培养6小时细胞发生差异表达的基因有37个,其中19个基因表达下调,18个基因表达上调。辐照后培养24小时细胞发生差异表达的基因有269个,202个基因表达下调,67个基因表达上调。这些基因大多与细胞信号转导、细胞周期调控、DNA损伤修复、细胞凋亡等相关。实验结果为阐明重离子辐射效应的分子机制提供了有用信息。
在进行上述实验研究的同时,学习和掌握了辐射致DNA和细胞损伤的几种检测技术,并对它们的适用性和优缺点进行了评估。
Proton and heavy ions have been applied in cancer radiation therapy field, and people have accepted this method increasingly. Under this background, the research on the biological effects and mechanisms of mammalian cell irradiated by high linear energy transfer (LET) ions has become more and more important.
This research investigate the biological effects of plasmid DNA and several cells after irradiation by methods of agarose gel electrophoresis along with advanced gel imaging system, colony forming ability experiment along with other traditional cell biology techniques and gene chip as a modern molecular biotechnology. The experimental results of this research can be expected to offer valuable basic data for cancer therapy by heavy ions and protons radiation method, Boron Neutron Capture Therapy (BNCT) as well as the dangerous assessment and protection of human in space or nuclear radiation environment, etc. The experimental procedure and elementary results are as follows:
The pUC19 plasmid DNA with or without free radical scavengers in aqueous solution is irradiated by 60Co-γrays, the results are analyzed by gel electrophoresis and Alpha Innotech digital imaging system .The results indicate that free radicals produced byγrays radiation are an important factor for the induction of DNA single strand breaks (SSB) and DNA double strand breaks (DSB). Mannitol, Vitamin C and tea polyphenols are all good scavengers, and mannitol is more effective than Vitamin C under the same condition. Compared with heavy ions, the capacity of scavengers is more remarkable on free radicals induced byγrays, which is more important to evaluate the effect of free radicals in heavy ion radiation.
U251 glioma cell and HepG2 liver cancer cell are irradiated by 60Co-γrays andαparticles emitted from 238Puαsource respectively. The results indicate that U251 cell is more sensitive to ionizing radiation than HepG2 cell, andαparticles irradiation inhibit the cell proliferation more effectively thanγrays irradiation. It means thatαparticles have a higher relative biological effectiveness (RBE) thanγrays.
L02 normal liver cell is irradiated by 5Gy 80MeV/u 12C6+ ions in radiation terminal of Heavy Ion Research Facility at Lanzhou (HIRFL). 22k human genome oligonucleotide microarray was used to explore the transcriptional profiles of the cells at 6h and 24h post-irradiation. In the 6h post-irradiation cells, the microarrays displayed 37 differentially expressed genes, including 18 up-regulated genes and 19 down-regulated genes. In the 24h post-irradiation cells, the microarrays displayed 269 differentially expressed genes, including 67 up-regulated gene and 202 down-regulated genes. Most of these changed genes were associated with signal transduction,cell cycle control,DNA repair and apoptosis. These observations provided the information of radiation-induced genes for elucidating molecular mechanisms of the diverse biological effects generated by heavy ion irradiation.
During the experiment mentioned above, several testing techniques on DNA and cell damage have been learnt and the adaptability,advantages and disadvantages of these methods are evaluated.
引文
[1] 复旦大学、清华大学和北京大学合编。原子核物理实验方法(上)。北京:原子能出版社,1985 年,第二版,48。
[2] 卫增泉。重离子束治癌,核物理动态,1994,11(3): 44-49。
[3] 杨垂绪,梅曼彤著。空间辐射生物学。广州:中山大学出版社,1995 年。
[4] 刘树铮。医学放射生物学,北京,原子能出版社,1998
[5] 李强,卫增泉。重离子束用于肿瘤放射治疗的基础理论,原子能物理评论,1999,16(4),262-266。
[6] 李强,卫增泉,党秉荣,等。重离子 Bragg 峰的研究及其应用,高能物理与核物理,1997,21(6),571-575
[7] 卫增泉,刘玉岩,王桂玲,等。46.6MeV/u 12C6+离子对四种作物种子的生物学效应研究,核物理动态,1993,10(3): 61-64。
[8] 夏寿萱,陈家佩,金璀珍,等.放射生物学.北京:军事医学科学出版社,1998 年,第一版。
[9] 夏寿萱,魏康,章杨培,等。分子放射生物学。北京:原子能出版社,1992 年,第一版。
[10] A. H. Sparrow, A. G. Underbrink, Rhoda C. Sparrow,et al.Chromosomes and Cellular Radiosensitivity.Radiation Research,1967,32(4):915-945
[11] Kraft. GSI-report, Darmstadt,Gesellschaft für Schwerionenforschung mbH, 1983: 201.
[12] Ager DD, Phillips JW, Columna EA, et al. Analysis of restriction enzyme- induced DNA double-strand breaks in Chinese hamster ovary cells by pulsed- field gel electrophoresis: implications for chromosome damage. Radiat Res, 1991,128: 150-156.
[13] Sutherland BM, Bennett PV a,Sutherland JC. Double strand breaks induced by low doses of γ rays or heavy ions: quantitation in nonradioactive human DNA. Anal Biochem, 1996,239: 53-60
[14]Daza P, Schuβler H, Mcmilian T J, et al. Radiosensitivity and double-strand break rejoining in tumorigenic and non-tumorigenic human epithelial cell lines. Int J Radiat Biol, 1997,72(1): 91-100
[15]Weber K J,Flentje M. Lethality of heavy ion-induced DNA double-strand breaks in mammalian cells. Int J Radiat Biol, 1993,64(2): 169-178
[16] D. Frankenberg DT Goodhead,M.Frankenberg-Schwager,et al. Effectiveness of 1.5 keV aluminum K and 0.3 keV carbon K characteristic X-rays at inducing DNA double-strand breaks in yeast cells, Int.J.Radiat.Biol., 1986, 50: 727-74
[17] M.O. Bradley ,Kohn KW. X-ray induced DNA double strand break production and repair in mammalian cells as measured by neutral filter elution, Nucleic Acids. Res., 1979, 7: 793-804
[18]李永明,赵玉琪。实用分子生物学方法手册。北京:科学出版社,1998年,第一版,1。
[19] J.M.沃克,R.拉普勒。分子生物学与实验技术。北京:化学工业出版社,2003年,第一版,21-22。
[20]C. Herskind et al., Single-strand breaks can lead to complex configuration of plasmid DNA in vitro, Int.J.Radiat.Biol., 1987, 52: 565-575
[21]田芳。原子力显微镜及其对DNA大分子的应用研究,物理,1997,26(4):238-243
[22]贺玉香。“彗星”分析法在放射生物学中的应用。广东医学,2003,24(11):1255-1257
[23]冯淑波,陈萍。 高效毛细管电泳的应用。保定师范专科学校学报,2006(2):31-33
[24]陈成忠,于洪芹。荧光原位杂交技术及其应用。生物学教学,2007(1):2-3
[25]曹建平,马岛秀行,山口千鹤,等。应用脉冲场凝胶电泳分析X射线诱发人骨肉瘤细胞DNA双链断裂与损伤修复效应。中华放射医学与防护杂志,2000,20(3):159-163
[26] 司徒镇强,吴军正。细胞培养。西安:世界图书出版西安公司,2004年:246-254
[27] 韩玲, 李雨, 陈杞,等。应用DNA 芯片技术结合核技术对辐射致癌分子机制的研究。国外医学·放射医学核医学分册,2001,25(2):78-81
[28] 滕霞,孙存谱。细胞凋亡和自由基。Chin J Ind Hyg Occup Dis, 1997,15(3):190-192
[29] Troy CM,Shelanski,ML.Down-regulation of copper/zinc superoxide dismutase cause apoptotic death in PC12 neuronal cells. Proc Natl Acad Sci USA, 1994, 91:6348-6387
[30] 张连琴,陈雪娴。自由基与疾病。天津医科大学学报,1997,3(2):85-88
[31] 任颖,刘兰静,王秋玉。自由基与衰老。长春中医学院学报,1997,13(62):59 [32 郑永裕。自由基反应与衰老,漳州职业大学学报,2003,2:113-115
[33] 高文庚。自由基理论在营养保健研究中的作用,运城学院生命科学系,2003,21(5):42-43
[34] 陈裕盛,郑颜萍。茶多酚抗氧自由基作用,中国肾脏病杂志,1998,14(1):65-68
[35] 林一萍,吴瑞荣。茶多酚在体外清除含氧自由基的作用分析,食品科学,2001,22(11):21-22
[36] 蒋建伟,何文珊,严玉霞。茶多酚的离体抗氧化作用 。中国病理生理杂志,1996,15(6):522-524
[37] 吴芹,杨郸丽,黄燮南.。甘露醇对羟自由基所致乳头肌收缩性能异常的保护作用。遵义医学院学报,1999,22(1):18-20
[38]马爱国,Susan J Duthie ,Marion A Ross,等。维生素 E、C 和 β-胡萝卜素对 DNA 损伤的影响中华预防医学杂志,1999,33(1):16-17
[39]傅春玲,江伟威,张萍,等。 维生素 C 抗辐射损伤作用研究。苏州医学院学报,2000,20(9):793-795
[40]蔡志强,赵希岳,李亮,等。三种天然抗氧化剂清除自由基作用的研究。中国药科大学学报,2004,35(2):190-192
[41]隋丽,赵葵,倪嵋楠,等. 7Li 和 12C 离子致 DNA 链断裂的研究.高能物理与核物理,2004,28(10):1126-1130
[42] SUI Li, ZHAO kui, NI Meinan ,et al. AFM Measurement of DNA Fragment Induced by Heavy Ions Chin. Phys. Lett. (in China), 2005,22(4):1010-1013
[43]孔福全,赵葵,展永,等。 随机断裂模型对7Li离子致DNA链断裂碎片长度分布的分析。科学通报,2005, 50 (10): 947-951
[44]赵葵,隋丽,孔福全,等。高 LET 的 7Li 离子致 DN 损伤的直接和间接作用研究。核技术,2007,30(4):250-254
[45] 冯惠茹,田嘉禾,张锦明。32P 照射对 Hela 细胞杀伤效应的研究.辐射研究与辐射工艺学报,2001,19(4):311-314
[46]Jones G D ,Milligan J R ,Ward J F, et al.Yield of strand breaks as a function of scavenger concentration and LET for SV40 irradiated with 4He ions.Radiation .Res. 1993,136:190-196
[47]Pang D, Rogers J E ,Berman B L,et al.Spatial distribution of radiation –induced double-strand breaks in plasmid DNA as resolved by atomic force microscopy. Radiat. Res.2005,164 :755-765
[48]蔡明辉,赵葵,展永,等。硼中子俘获治疗癌症的基础性实验研究。原子能科学技术,2006,40(4):391—395。
[49] 王菊芳,周光明,何静,等。不同LET碳离子对V79 细胞辐射敏感性的影响。辐射研究与辐射工艺学报,2001,19 (1):36-39
[50]Zirkle RE, Marchbank DF, Kuck DD.Exponential and sigmoid survival curves resulting from alpha and X-irradiation of Aspergillus spores[J]. J Cell Comp Physiol, 1952, 39:75.
[51] Barendsen G W , Walter H M D , Fowler J P ,et al . Radiat Res , 1963 , 18 (2) :106 - 199
[52]Todd P W. Radiat Res , 1967 , 7 : S196 - S207
[53]Weisbrod U , Bucker H , Horneck G ,et al . Radiat Res , 1992 , 129 (3) : 250 - 257
[54]Thacker J , Stretch A , Stephens A. Int J Radiol Biol , 1979 , 36 (2) : 137 - 148
[55] Furusawa Y, Fukutsu K, Ando K. Biological effectiveness of heavy - ion beams on cultured mammalian cells.RIKEN Accelerator Progress Report 1997
[56]]Fornace AJ Jr, Amundson SA, Bittner M, et al. The complexity of radiation stress responses: analysis byinformatics and functional genomics approaches [J]. Gene Expr , 1999,7(4-6):387-400.
[57]Schena M, Shalon D, Davis R.W, et al. Quantitative monitoring of geneexpression patterns with a complementary DNA microarray [J]. Science, 1995. 270: 467- 470.
[58] Epstein CB, Butow R. Microarray technology-enhanced versatility, persistent challenge[J].Curr Opin Biotechnol,2000,11(1):36-41
[59] Amundson SA ,Bittner M, Meltzer P, et al. Induction of gene expression as a monitor of exposure to ionizing radiation [J]. Radiat Res, 2001, 156(5 Pt2): 657-661
[60]高荣莲,陈肖华,饶亚岚,等。寡聚核苷酸芯片技术筛选电离辐射后大鼠 PC12 细胞的差异表达基因。中华放射医学与防护杂志,2005,25 (3):242-245
[61]林琼燕。核转录因子кB 系统与肿瘤放射敏感性。广东医学院学报,2005,23(1):84-85
[62]铁轶,罗瑛,隋建丽,等。辐射诱导基因的寻找及功能研究。中华放射医学与防护杂志,2000,20(6 ):446-448
[63]郭万峰,林汝仙,黄坚,等。辐射相关基因芯片的制备及其在肺癌细胞中的应用。中华放射医学与防护杂志,2005,25(2):107-110
[64]韩玲, 李雨, 陈杞,等。应用DNA 芯片技术结合核技术对辐射致癌分子机制的研究。国外医学·放射医学核医学分册,2001,25(2):78-80
[65]饶亚岚,陈肖华,毛秉智,等。DNA 微阵列技术在辐射诱导基因表达研究中的运用。辐射研究与辐射工艺学报,2003,21(1):25-28
[66]李雨,李瑶。基因芯片技术在放射生物学研究领域的应用。国外医学·放射医学核医学分册,2002,26(4):177-179
[2] 卫增泉。重离子束治癌,核物理动态,1994,11(3): 44-49。
[3] 杨垂绪,梅曼彤著。空间辐射生物学。广州:中山大学出版社,1995 年。
[4] 刘树铮。医学放射生物学,北京,原子能出版社,1998
[5] 李强,卫增泉。重离子束用于肿瘤放射治疗的基础理论,原子能物理评论,1999,16(4),262-266。
[6] 李强,卫增泉,党秉荣,等。重离子 Bragg 峰的研究及其应用,高能物理与核物理,1997,21(6),571-575
[7] 卫增泉,刘玉岩,王桂玲,等。46.6MeV/u 12C6+离子对四种作物种子的生物学效应研究,核物理动态,1993,10(3): 61-64。
[8] 夏寿萱,陈家佩,金璀珍,等.放射生物学.北京:军事医学科学出版社,1998 年,第一版。
[9] 夏寿萱,魏康,章杨培,等。分子放射生物学。北京:原子能出版社,1992 年,第一版。
[10] A. H. Sparrow, A. G. Underbrink, Rhoda C. Sparrow,et al.Chromosomes and Cellular Radiosensitivity.Radiation Research,1967,32(4):915-945
[11] Kraft. GSI-report, Darmstadt,Gesellschaft für Schwerionenforschung mbH, 1983: 201.
[12] Ager DD, Phillips JW, Columna EA, et al. Analysis of restriction enzyme- induced DNA double-strand breaks in Chinese hamster ovary cells by pulsed- field gel electrophoresis: implications for chromosome damage. Radiat Res, 1991,128: 150-156.
[13] Sutherland BM, Bennett PV a,Sutherland JC. Double strand breaks induced by low doses of γ rays or heavy ions: quantitation in nonradioactive human DNA. Anal Biochem, 1996,239: 53-60
[14]Daza P, Schuβler H, Mcmilian T J, et al. Radiosensitivity and double-strand break rejoining in tumorigenic and non-tumorigenic human epithelial cell lines. Int J Radiat Biol, 1997,72(1): 91-100
[15]Weber K J,Flentje M. Lethality of heavy ion-induced DNA double-strand breaks in mammalian cells. Int J Radiat Biol, 1993,64(2): 169-178
[16] D. Frankenberg DT Goodhead,M.Frankenberg-Schwager,et al. Effectiveness of 1.5 keV aluminum K and 0.3 keV carbon K characteristic X-rays at inducing DNA double-strand breaks in yeast cells, Int.J.Radiat.Biol., 1986, 50: 727-74
[17] M.O. Bradley ,Kohn KW. X-ray induced DNA double strand break production and repair in mammalian cells as measured by neutral filter elution, Nucleic Acids. Res., 1979, 7: 793-804
[18]李永明,赵玉琪。实用分子生物学方法手册。北京:科学出版社,1998年,第一版,1。
[19] J.M.沃克,R.拉普勒。分子生物学与实验技术。北京:化学工业出版社,2003年,第一版,21-22。
[20]C. Herskind et al., Single-strand breaks can lead to complex configuration of plasmid DNA in vitro, Int.J.Radiat.Biol., 1987, 52: 565-575
[21]田芳。原子力显微镜及其对DNA大分子的应用研究,物理,1997,26(4):238-243
[22]贺玉香。“彗星”分析法在放射生物学中的应用。广东医学,2003,24(11):1255-1257
[23]冯淑波,陈萍。 高效毛细管电泳的应用。保定师范专科学校学报,2006(2):31-33
[24]陈成忠,于洪芹。荧光原位杂交技术及其应用。生物学教学,2007(1):2-3
[25]曹建平,马岛秀行,山口千鹤,等。应用脉冲场凝胶电泳分析X射线诱发人骨肉瘤细胞DNA双链断裂与损伤修复效应。中华放射医学与防护杂志,2000,20(3):159-163
[26] 司徒镇强,吴军正。细胞培养。西安:世界图书出版西安公司,2004年:246-254
[27] 韩玲, 李雨, 陈杞,等。应用DNA 芯片技术结合核技术对辐射致癌分子机制的研究。国外医学·放射医学核医学分册,2001,25(2):78-81
[28] 滕霞,孙存谱。细胞凋亡和自由基。Chin J Ind Hyg Occup Dis, 1997,15(3):190-192
[29] Troy CM,Shelanski,ML.Down-regulation of copper/zinc superoxide dismutase cause apoptotic death in PC12 neuronal cells. Proc Natl Acad Sci USA, 1994, 91:6348-6387
[30] 张连琴,陈雪娴。自由基与疾病。天津医科大学学报,1997,3(2):85-88
[31] 任颖,刘兰静,王秋玉。自由基与衰老。长春中医学院学报,1997,13(62):59 [32 郑永裕。自由基反应与衰老,漳州职业大学学报,2003,2:113-115
[33] 高文庚。自由基理论在营养保健研究中的作用,运城学院生命科学系,2003,21(5):42-43
[34] 陈裕盛,郑颜萍。茶多酚抗氧自由基作用,中国肾脏病杂志,1998,14(1):65-68
[35] 林一萍,吴瑞荣。茶多酚在体外清除含氧自由基的作用分析,食品科学,2001,22(11):21-22
[36] 蒋建伟,何文珊,严玉霞。茶多酚的离体抗氧化作用 。中国病理生理杂志,1996,15(6):522-524
[37] 吴芹,杨郸丽,黄燮南.。甘露醇对羟自由基所致乳头肌收缩性能异常的保护作用。遵义医学院学报,1999,22(1):18-20
[38]马爱国,Susan J Duthie ,Marion A Ross,等。维生素 E、C 和 β-胡萝卜素对 DNA 损伤的影响中华预防医学杂志,1999,33(1):16-17
[39]傅春玲,江伟威,张萍,等。 维生素 C 抗辐射损伤作用研究。苏州医学院学报,2000,20(9):793-795
[40]蔡志强,赵希岳,李亮,等。三种天然抗氧化剂清除自由基作用的研究。中国药科大学学报,2004,35(2):190-192
[41]隋丽,赵葵,倪嵋楠,等. 7Li 和 12C 离子致 DNA 链断裂的研究.高能物理与核物理,2004,28(10):1126-1130
[42] SUI Li, ZHAO kui, NI Meinan ,et al. AFM Measurement of DNA Fragment Induced by Heavy Ions Chin. Phys. Lett. (in China), 2005,22(4):1010-1013
[43]孔福全,赵葵,展永,等。 随机断裂模型对7Li离子致DNA链断裂碎片长度分布的分析。科学通报,2005, 50 (10): 947-951
[44]赵葵,隋丽,孔福全,等。高 LET 的 7Li 离子致 DN 损伤的直接和间接作用研究。核技术,2007,30(4):250-254
[45] 冯惠茹,田嘉禾,张锦明。32P 照射对 Hela 细胞杀伤效应的研究.辐射研究与辐射工艺学报,2001,19(4):311-314
[46]Jones G D ,Milligan J R ,Ward J F, et al.Yield of strand breaks as a function of scavenger concentration and LET for SV40 irradiated with 4He ions.Radiation .Res. 1993,136:190-196
[47]Pang D, Rogers J E ,Berman B L,et al.Spatial distribution of radiation –induced double-strand breaks in plasmid DNA as resolved by atomic force microscopy. Radiat. Res.2005,164 :755-765
[48]蔡明辉,赵葵,展永,等。硼中子俘获治疗癌症的基础性实验研究。原子能科学技术,2006,40(4):391—395。
[49] 王菊芳,周光明,何静,等。不同LET碳离子对V79 细胞辐射敏感性的影响。辐射研究与辐射工艺学报,2001,19 (1):36-39
[50]Zirkle RE, Marchbank DF, Kuck DD.Exponential and sigmoid survival curves resulting from alpha and X-irradiation of Aspergillus spores[J]. J Cell Comp Physiol, 1952, 39:75.
[51] Barendsen G W , Walter H M D , Fowler J P ,et al . Radiat Res , 1963 , 18 (2) :106 - 199
[52]Todd P W. Radiat Res , 1967 , 7 : S196 - S207
[53]Weisbrod U , Bucker H , Horneck G ,et al . Radiat Res , 1992 , 129 (3) : 250 - 257
[54]Thacker J , Stretch A , Stephens A. Int J Radiol Biol , 1979 , 36 (2) : 137 - 148
[55] Furusawa Y, Fukutsu K, Ando K. Biological effectiveness of heavy - ion beams on cultured mammalian cells.RIKEN Accelerator Progress Report 1997
[56]]Fornace AJ Jr, Amundson SA, Bittner M, et al. The complexity of radiation stress responses: analysis byinformatics and functional genomics approaches [J]. Gene Expr , 1999,7(4-6):387-400.
[57]Schena M, Shalon D, Davis R.W, et al. Quantitative monitoring of geneexpression patterns with a complementary DNA microarray [J]. Science, 1995. 270: 467- 470.
[58] Epstein CB, Butow R. Microarray technology-enhanced versatility, persistent challenge[J].Curr Opin Biotechnol,2000,11(1):36-41
[59] Amundson SA ,Bittner M, Meltzer P, et al. Induction of gene expression as a monitor of exposure to ionizing radiation [J]. Radiat Res, 2001, 156(5 Pt2): 657-661
[60]高荣莲,陈肖华,饶亚岚,等。寡聚核苷酸芯片技术筛选电离辐射后大鼠 PC12 细胞的差异表达基因。中华放射医学与防护杂志,2005,25 (3):242-245
[61]林琼燕。核转录因子кB 系统与肿瘤放射敏感性。广东医学院学报,2005,23(1):84-85
[62]铁轶,罗瑛,隋建丽,等。辐射诱导基因的寻找及功能研究。中华放射医学与防护杂志,2000,20(6 ):446-448
[63]郭万峰,林汝仙,黄坚,等。辐射相关基因芯片的制备及其在肺癌细胞中的应用。中华放射医学与防护杂志,2005,25(2):107-110
[64]韩玲, 李雨, 陈杞,等。应用DNA 芯片技术结合核技术对辐射致癌分子机制的研究。国外医学·放射医学核医学分册,2001,25(2):78-80
[65]饶亚岚,陈肖华,毛秉智,等。DNA 微阵列技术在辐射诱导基因表达研究中的运用。辐射研究与辐射工艺学报,2003,21(1):25-28
[66]李雨,李瑶。基因芯片技术在放射生物学研究领域的应用。国外医学·放射医学核医学分册,2002,26(4):177-179