【作者】 朱巍；

【导师】 郑斯英；

【作者基本信息】 苏州大学 ， 放射医学， 2003， 硕士

【摘要】 目的：探索用Calyculin A诱导外周血淋巴细胞染色体发生凝聚的最优化方法，研究Calyculin A诱导的染色体凝聚与辐射剂量之间的剂量效应关系，探讨Calyculin A诱导染色体凝聚在临床上的应用。 方法 采用L₉（3⁴）正交试验，对影响Calyculin A诱导染色体凝聚方法3个主要影响因素（Calyculin A的终浓度，Calyculin A的处理时间，细胞培养时间）进行探讨。采用不同剂量（0、0.25、0.5、1.0、2.0、3.0Gy）的⁶⁰Coγ射线照射离体人血，观察其诱导的染色体凝聚，并与常规染色体畸变法进行比较。并采用Calyculin A诱导的染色体凝聚断片率来监测甲状腺癌患者接受¹³¹Ⅰ治疗过程中染色体损伤程度。 结论 Calyculin A的终浓度，Calyculin A的处理时间，细胞培养时间不同水平的影响具有统计学意义（P＜0.01）。淋巴细胞培养48h，培养结束前2h加入Calyculin　A，Calyculin　A终浓度为50nmol/L的实验组合可以获得高比例的分裂指数。Calyculin A法染色体凝聚断片率与外照射剂量之间有良好的线性关系。断片率与剂量之间的关系可用线性关系式表示：Y＝10.405+0.358X，相关系数r＝0.9926，其中X为γ线外照射剂量（cGy），Y为Calyculin A法PCC断片率（％）。Calyculin A诱导的染色体凝聚断片率比常规染色体双着丝粒体率更真实反映甲状腺癌患者接受’3’I治疗过程中染色体损伤程度.更多还原

【Abstract】 Objective To explore the best method to assay the PCC (premature chromosome condensation) of human peripheral blood lymphocytes induced by CalyculinA and study the effect-dosage relationship between the PCC yield in peripheral blood lymphocytes exposed to Calyculin A and the IR absobed dosage ,to verify the prospective applicability of the method in the clinical trial as a a quantitative biodosimeter.Methods We employed the designed L9(34) Orthogonal test and studided the three main factors that influcing the mothod of PCC induced by Calyculin A, including the concentration of Calyculin A,the time passed after the exposure to Calyculin A and the cells we observed. The experiments were carried out on human peripheral blood lymphocytes in vitro exposed to 60Co y -rays. The doses absorbed amounted to 0, 0.25, 0.5, 1.0, 2.0, 3.0Gy, we observed the premature chromosome condensation following the radiation, then compared the results with the general chromosome aberration assay technique. We also applied the method to assess the chromosome aberrations frequencies of the thyroid cancer patients treated with Iodine 131 radiotherapy, To verify the applicability of the PCC yield in peripheral blood lymphocytes as a quantitative biodosimeter.Conclusion We observed that different concentration of Calyculin A, treatment time and cell culture time result in different PCC yield in the lymphocytes. After lymphocytes were cultured for 48h ,Calyculin A was added to the medium (up to 50nmol/L) 2 hours before the end of culture. In that case we can get the highest ratio cytokinesis cells. The fragment rate of chromosome condensation induced by Calyculin A has a favorable linearity relationship with external radiation dose. The linearity relationship Y = 10.405 + 0.358 X .Relationship coefficient r = 0.9926 ,in which X is external radiation dose (cGy) ,Y is fragment rate of PCC(%) induced by Calyculin A. We found that in the assessment of chromosome damage during the process of radiotherapy using iodin-131 on thyroid cancer patients, the fragment rate of chromosome condensation induced by Calyculin A was more precise than general chromosome assay technique.更多还原