Calyculin A诱导早熟染色体与低剂量电离辐射剂量-效应关系
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摘要
目的探讨CalyculinA诱导早熟染色体凝集技术作为低剂量电离辐射生物剂量计的可行性。方法取健康成年人的抗凝外周静脉血,用X射线分别以0、0.1、0.25、0.5、0.75和1.0Gy照射。照射后将血样接种于RP-MI1640培养基中,于37℃、5%CO2恒温培养箱内培养48h,当培养24h时加入秋水仙素,终止培养前2h加入Calyculi-nA诱导早熟凝集染色体,收获制片并用Giemsa染液染色,观察G1、G2/M-PCC细胞中总畸变率、断片率、双着丝粒体加着丝粒环率与辐射剂量的效应关系。结果用CalyculinA可成功诱导人外周血淋巴细胞产生PCC,且总畸变率、断片率、双着丝粒体加着丝粒环率随照射剂量的增加而增加,最佳拟合曲线为二次多项式模式。结论 CalyculinA诱导PCC技术可以作为一种方法来估算低剂量受照者的辐射剂量,但是需要更多的资料来完善。
Objective To explore the feasibility of CalyculinA induced premature chromosome condensation to be used as a biodosimetry of low-dose ionizing radiation.Methods Peripheral blood was drawn from healthy adults and Anticoagulated with Heparin.a set of tubes was irradiated by X-ray.The irradiation dose included 0,0.1,0.25,0.5,0.75,1.0Gy.After irradiation,the whole blood was added to each RPMI1640 medium,All incubations were Cultured for up to 48h at 37℃ in an atmosphere of 5% CO2 in air.Colcemid was added 24hours after cultures start and Calyculin A was added for the last 2 hours to induce PCC.The chromosome spreads were prepared,stained with Giemsa and observed under a microscope.The dose-response relationship of the total aberration,fragment,dicentric+centric ring(dic+r) in the irradiated lymphocytes was examined.Results The PCC was successfully induced in lymphocytes with Calyculin A.The yield of chromosome aberrations increased with the dose going up.An aquadratic polynomial dose response model was fitted.Conclusion Premature chromosome condensation induced by Calyculin A can be used as a technology to estimate the radiation dosage of low-dose ionizing radiation,but still need more data to improve.
Objective To explore the feasibility of CalyculinA induced premature chromosome condensation to be used as a biodosimetry of low-dose ionizing radiation.Methods Peripheral blood was drawn from healthy adults and Anticoagulated with Heparin.a set of tubes was irradiated by X-ray.The irradiation dose included 0,0.1,0.25,0.5,0.75,1.0Gy.After irradiation,the whole blood was added to each RPMI1640 medium,All incubations were Cultured for up to 48h at 37℃ in an atmosphere of 5% CO2 in air.Colcemid was added 24hours after cultures start and Calyculin A was added for the last 2 hours to induce PCC.The chromosome spreads were prepared,stained with Giemsa and observed under a microscope.The dose-response relationship of the total aberration,fragment,dicentric+centric ring(dic+r) in the irradiated lymphocytes was examined.Results The PCC was successfully induced in lymphocytes with Calyculin A.The yield of chromosome aberrations increased with the dose going up.An aquadratic polynomial dose response model was fitted.Conclusion Premature chromosome condensation induced by Calyculin A can be used as a technology to estimate the radiation dosage of low-dose ionizing radiation,but still need more data to improve.
引文
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[7]王春燕,佟鹏,苟巧,等.用早熟凝集染色体环法研究中子诱发染色体畸变的剂量效应关系[J].畸变、癌变、突变,2011,23:449-451.
[8]George K,Durante M,Wu H,et al.In vivo and in vitromeasurements of complex-type chromosomal exchanges in-duced by heavy ions[J].Adv Space Res,2003,31:1 525-1 535.
[9]白玉书,黄绮龙,马剑锋,等.高剂量率射线照射离体血建立染色体畸变的剂量-效应曲线[J].中国预防医学杂志,2003,4(2):100-103.
[2]Wang ZZ,Li WJ,Zhi DJ,et al.Biodosimetry estimate forhigh-LET irradiation[J].Radiat Environ Biophys,2007,46:229-35.
[3]Gotoh E,Kawata T,Durante M.Chromatid break rejoiningand exchange aberration formation following gamma-ray ex-posure:analysis in G2 human fibroblasts by chemically in-duced premature chromosome condensation[J].Radiat Biol,1999,75(9):1 129-1 135.
[4]Kawata T,Durante M,Furusawa Y,et al.Dose-response ofinitial G2-chromatid breaks induced in normal human fibro-blasts by heavy ions[J].Int J Radiat Biol,2001,77(2):165-174.
[5]Balakrishnan S,Shirsath K,Bhat N,et al.Biodosimetry forhigh dose accidental exposures by drug induced prematurechromosome condensation(PCC)assay[J].Mutat Res.2010,699:11-16.
[6]Lamadrid AI,Garcia O,Delbos M,et al.PCC-ring Induc-tion in Human Lymphocytes Exposed to Gamma and NeutronIrradiaton[J].Radiat.Res,2007,48:1-6.
[7]王春燕,佟鹏,苟巧,等.用早熟凝集染色体环法研究中子诱发染色体畸变的剂量效应关系[J].畸变、癌变、突变,2011,23:449-451.
[8]George K,Durante M,Wu H,et al.In vivo and in vitromeasurements of complex-type chromosomal exchanges in-duced by heavy ions[J].Adv Space Res,2003,31:1 525-1 535.
[9]白玉书,黄绮龙,马剑锋,等.高剂量率射线照射离体血建立染色体畸变的剂量-效应曲线[J].中国预防医学杂志,2003,4(2):100-103.