^(60)Co γ射线照射正常人淋巴母细胞诱导CDKN1A基因mRNA表达水平的变化

Radiation Effects of ^(60)Co γ-Rays on Expression of CDKN1A mRNA in Human Lymphocytoblast

目的探讨不同剂量60Coγ射线在不同培养时间后对人淋巴母细胞AHH-1细胞CDKN1A基因mRNA表达的影响。方法用不同剂量(0、0.2、1、3、5、10Gy)的60Coγ射线照射正常人淋巴母细胞AHH-1细胞,在不同维持存活培养时间(0、4、24、48、72、96 h)内收集细胞,抽提总RNA,用实时PCR方法检测细胞中CDKN1A基因mRNA表达水平,观察其随辐照剂量和培养时间的变化。结果 AHH-1细胞中CDKN1A基因mRNA的表达水平随辐照剂量的加大而增加,在0～5 Gy之间具有一定剂量依赖性关系(P<0.05);辐照后培养24 h时基因表达达到峰值,24 h之后呈现下降趋势。结论不同剂量γ射线照射人AHH-1细胞会导致CDKN1A基因表达水平在培养24 h内随辐射剂量增加而升高,这种剂量依赖性关系可能适用于电离辐射剂量的估算。

Objective To study effect of 60Co γ-rays radiation with different doses on expression of CDKN1A gene mRNA in human lymphocytoblast cultured for different time.Methods After human lymphocytoblasts were irradiated by the 60Co γ-rays with various doses of 0,0.2,1,3,5 and 10 Gy,and cells were separately cultured for sustaining survival during 0,4,24,48,72 and 96 h.The total RNA was extracted from each sample and the real-time PCR was conducted to observe the gene CDKN1A mRNA level changes in lymphocytoblasts exposed to various radiation doses for various cultured time.Results Expressive levels of the CDKN1A mRNA in lymphocytoblasts gradually went up with increasing radiation doses,which showed γ-rays dose dependent from 0 Gy to 5 Gy（P0.05）,and reach the peak when cells were cultured for 24 h after exposing to radiation while displayed an expressive downtrend during the later stage of cell culture.Conclusion Increase of the CDKN1A mRNA expression level in human lymphocytoblasts after exposing to 60Co γ-rays radiation within 24 hours of culture shows a dose dependent way,which may be used to evaluate the ionizing radiation dose.