Marples B, Longhurst D, Eastham A M, West C M
Cancer Research Campaign Department of Experimental Radiation Oncology, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Manchester, UK.
Br J Cancer. 1998 Apr;77(7):1108-14. doi: 10.1038/bjc.1998.184.
The single-cell gel electrophoresis (comet) assay was used to measure radiation-produced DNA double-strand breaks (dsbs) in a series of seven cervical tumour cell lines (ME180, HT3, C33A, C41, SiHa, MS751 and CaSki). The proportion of DNA dsbs was measured immediately after radiation treatment (initial damage) and 16 h later after incubation at 37 degrees C (residual damage). Linear dose-response curves were seen for initial (slopes 0.23-0.66) and residual (slopes 0.16-0.87) DNA dsbs. Neither of the slopes of the linear regression analysis on the initial and on the residual DNA dsbs dose-response curves (range 0-80 Gy) correlated with SF2 (surviving fraction at 2 Gy) measured after high- (HDR) or low-dose-rate (LDR) irradiation. An association was evident between SF2 after HDR and LDR irradiation and the ratio of the absolute level of initial and residual damage after a single dose of 60 Gy. However, a significant correlation was found between HDR (r= -0.78, P = 0.04) and LDR (r = -0.86, P = 0.03) SF2 values and the ratio of the slopes of the initial and residual DNA dsbs dose-response curves (range 0.47-0.99), representing the fraction of DNA damage remaining. These results indicate that the neutral comet assay can be used to predict radiosensitivity of cervical tumour cell lines by assessing the ratio of initial and residual DNA dsbs.
采用单细胞凝胶电泳(彗星)试验,检测一系列7种宫颈肿瘤细胞系(ME180、HT3、C33A、C41、SiHa、MS751和CaSki)中辐射产生的DNA双链断裂(dsbs)。在放射治疗后立即(初始损伤)以及在37℃孵育16小时后(残留损伤)测量DNA双链断裂的比例。观察到初始(斜率0.23 - 0.66)和残留(斜率0.16 - 0.87)DNA双链断裂的线性剂量反应曲线。在初始和残留DNA双链断裂剂量反应曲线(范围0 - 80 Gy)上进行的线性回归分析的斜率,均与高剂量率(HDR)或低剂量率(LDR)照射后测得的SF2(2 Gy时的存活分数)无关。HDR和LDR照射后的SF2与单剂量60 Gy后初始和残留损伤的绝对水平之比之间存在明显关联。然而,发现HDR(r = -0.78,P = 0.04)和LDR(r = -0.86,P = 0.03)的SF2值与初始和残留DNA双链断裂剂量反应曲线斜率之比(范围0.47 - 0.99)之间存在显著相关性,该斜率之比代表剩余的DNA损伤分数。这些结果表明,中性彗星试验可通过评估初始和残留DNA双链断裂的比例来预测宫颈肿瘤细胞系的放射敏感性。