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目的探讨γ射线辐射对人淋巴细胞平均端粒长度的影响。方法培养非洲淋巴细胞瘤病毒转化正常人B淋巴细胞(Peng-EBV细胞),接受钴-60γ射线照射,剂量率为0.96 Gy/min,吸收剂量分别为0.2、0.6、1.6、3.0和6.0 Gy;以不进行照射的细胞作为对照组。分别收集照射后24、48和72 h细胞样品,提取基因组DNA。采用定量聚合酶链式反应(Q-PCR)法检测样品平均端粒长度。分析γ辐射影响人淋巴细胞端粒长度的剂量-效应和时间-效应。结果端粒标准DNA的标准曲线为Ct=B+M×lg D=35.264-3.339 lg D(相关系数=0.964,P<0.05)。γ射线单次照射可导致Peng-EBV细胞在照射后24~72 h端粒长度较同时间点对照组延长(P<0.05)。在0.0~1.6 Gy剂量范围内,γ射线诱导的Peng-EBV细胞端粒长度在照射后48 h有随剂量增加而增加的趋势(P<0.05)。0.6和1.6 Gy 2个剂量组的端粒延长幅度有随照后时间延长而增大的趋势(P<0.05)。结论Q-PCR方法适用于淋巴细胞绝对端粒长度检测。γ射线单次照射可以导致Peng-EBV细胞平均端粒长度增加。
Objective To investigate the effect of γ-ray irradiation on the average telomere length of human lymphocytes. Methods The human lymphoblastoma virus was transformed into normal human B lymphocytes (Peng-EBV cells) and irradiated with cobalt-60γ ray at a dose rate of 0.96 Gy / min. The absorbed doses were 0.2, 0.6, 1.6, 3.0 and 6.0 Gy, respectively. The non-irradiated cells served as the control group. Cell samples were collected 24, 48 and 72 h after irradiation, and genomic DNA was extracted. The average telomere length was measured by quantitative polymerase chain reaction (Q-PCR). Analysis of dose-effect and time-effect of gamma irradiation on human lymphocyte telomere length. Results The standard curve of telomere standard DNA was Ct = B + M × lg D = 35.264-3.339 lg D (correlation coefficient = 0.964, P <0.05). The single irradiation of γ-ray could cause the telomere length of Peng-EBV cells to extend 24- 72 h after irradiation (P <0.05). In the dose range of 0.0-1.6 Gy, the telomere length of Peng-EBV cells induced by γ-irradiation increased with dose (P <0.05) at 48 h after irradiation. The telomere elongation of the two dose groups 0.6 and 1.6 Gy tended to increase with time (P <0.05). Conclusion Q-PCR method is suitable for the detection of absolute telomere length of lymphocytes. Single irradiation with gamma rays can result in an increase in the average telomere length of Peng-EBV cells.