早熟凝集染色体技术在核事故应用中的方法学改进  

Improvement of Method for the Technology of Premature Chromosome Condensation in Nuclear Accident

作　　者：郭义曹[1] 郑巧玲[1] 李禧娜[1] 孙杰[1] 谢国强[1] 廖国兰[1] 

机构地区：[1]广东省职业病防治院,广东广州510300

出　　处：《中国辐射卫生》2011年第3期275-277,共3页Chinese Journal of Radiological Health

基　　金：广东省医学科学技术研究基金资助项目(A2008060)

摘　　要：目的研究早熟凝集染色体(PCC)技术的方法学改进,为核和电离辐射事故时快速估算受照生物剂量提供良好的读片标本。方法取人的肘静脉血,以常规的PCC培养方法为标准,通过对培养中加入化学诱导剂花萼海绵体诱癌素A(CA)、有丝分裂阻断剂秋水仙素来研究收获PCC的形态和长度;通过对制片方法的改进来增强收获的PCC形态达到良好的状态;加入苦杏仁苷和细胞松驰素B(Cyt-B)2种细胞增殖促进剂在60Coγ射线大剂量照射的条件下,对促进周围血淋巴细胞增殖的作用。结果①以加入CA为35 nmol/L、秋水仙素30 nmol/L培养50 h后收获的PCC形态与结构最好,符合长度∶宽度比例要求的细胞率最高(47.4%);②制片中增加低渗液的用量至8 ml和延长时间至30 min并在37℃水浴下进行,同时改变固定液的比例为2∶1能增强收获良好的PCC标本;③照射0～20.0 Gy后,以质量浓度苦杏仁苷为150 mg/L和Cyt-B为0.2 mg/L共同培养收获的细胞增殖率最高,常规法培养最低。结论本研究的方法学改进可快速获得良好的PCC读片标本,为核和电离辐射事故时快速估算受照的生物剂量提供方便。Objective To study the improvement of method for the technology of premature chromosome condensation （PCC） ,and provide a good specimen for quickly estimating radiation dose in nuclear and ionizing accident emergency. Methods The conventional PCC method was used to study the morphology and length of PCC with chemical inducer and mitosis blocker, calyculin A（CA） and colchicines. Improved the dialysis method for harvesting a good morphology of PCC. 2 kinds of cell proliferation promoting agents were added after high dose ionizing radiation with ^60Coγ/-ray, amygdalin and Cyt -B, to promote the proliferation effects of human peripheral blood lymphocytes. Results （1）CA （35 nmol/L） and eolchicines （30 nmol/L） were added and cultivated for 50 hours. The PCC with best morphology and structure could be harvested. The cell rate of suitable length vs width was highest（47.4% ）. （2）The dose of hypotonic solution was increased, cultivated in 37 ℃ water bath and the ratio of fixing solution was changed. The PCC with good specimen could be harvested. （3）After being irradiated by 0 -20.0 Gy ^60Coγ/-ray, and the cells were cultivated for 50 hours with amygdalin（ 150 mg/L） and Cyt -B （0.2 mg/L）. The cell proliferation rate was the highest, while the rate in conventional method was lowest. Conclusion The improvement of method could rapidly harvest the good specimen of PCC,it is sutiable for quick evaluation of biological dose when ionizing radiation accident occurred.

关 键 词：早熟凝集染色体 方法学改进 核事故 快速 生物剂量估算 

分 类 号：Q345[生物学—遗传学]