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中国仓鼠卵巢细胞系AA8和EM9经365纳米紫外线A辐射后细胞存活与DNA单链断裂修复能力之间的相关性

Correlation between cell survival and DNA single-strand break repair proficiency in the Chinese hamster ovary cell lines AA8 and EM9 irradiated with 365-nm ultraviolet-A radiation.

作者信息

Churchill M E, Peak J G, Peak M J

机构信息

Molecular Photobiology Group, Argonne National Laboratory, IL 60439-4833.

出版信息

Photochem Photobiol. 1991 Feb;53(2):229-36. doi: 10.1111/j.1751-1097.1991.tb03927.x.

Abstract

Cell survival parameters and the induction and repair of DNA single-strand breaks were measured in two Chinese hamster ovary cell lines after irradiation with monochromatic UVA radiation of wavelength 365 nm. The radiosensitive mutant cell line EM9 is known to repair ionizing-radiation-induced single-strand breaks (SSB) more slowly than the parent line AA8. EM9 was determined to be 1.7-fold more sensitive to killing by 365-nm radiation than AA8 at the 10% survival level, and EM9 had a smaller shoulder region on the survival curve (alpha = 1.76) than AA8 (alpha = 0.62). No significant differences were found between the cell lines in the initial yields of SSB induced either by gamma-radiation (as determined by alkaline sucrose gradient sedimentation) or by 365-nm UVA (as determined by alkaline elution). For measurement of initial SSB, cells were irradiated at 0.5 degrees C to minimize DNA repair processes. Rejoining of 365-nm induced SSB was measured by irradiating cells at 0.5 degrees C, allowing them to repair at 37 degrees C in full culture medium, and then quantitating the remaining SSB by alkaline elution. The repair of these breaks followed biphasic kinetics in both cell lines. EM9 repaired the breaks more slowly (t1/2 values of 1.3 and 61.3 min) than did AA8 (t1/2 values of 0.9 and 53.3 min), and EM9 also left more breaks unrepaired 90 min after irradiation (24% vs 8% for AA8). Thus, the sensitivity of EM9 to 365-nm radiation correlated with its deficiency in repairing DNA lesions revealed as SSB in alkaline elution.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在用波长365nm的单色UVA辐射照射后,对两种中国仓鼠卵巢细胞系的细胞存活参数以及DNA单链断裂的诱导和修复情况进行了测定。已知放射敏感突变细胞系EM9修复电离辐射诱导的单链断裂(SSB)的速度比亲本细胞系AA8慢。在10%存活水平下,EM9对365nm辐射杀伤的敏感性比AA8高1.7倍,并且EM9在存活曲线上的肩部区域比AA8小(α = 1.76对α = 0.62)。在由γ辐射(通过碱性蔗糖梯度沉降测定)或365nm UVA(通过碱性洗脱测定)诱导的SSB的初始产率方面,细胞系之间未发现显著差异。为了测量初始SSB,细胞在0.5℃下照射以尽量减少DNA修复过程。通过在0.5℃下照射细胞、使其在37℃的完全培养基中修复,然后通过碱性洗脱对剩余的SSB进行定量,来测量365nm诱导的SSB的重新连接情况。在两种细胞系中,这些断裂的修复均遵循双相动力学。EM9修复断裂的速度比AA8慢(t1/2值分别为1.3和61.3分钟对0.9和53.3分钟),并且在照射90分钟后EM9未修复的断裂也更多(AA8为8%,EM9为24%)。因此,EM9对365nm辐射的敏感性与其在修复碱性洗脱中显示为SSB的DNA损伤方面的缺陷相关。(摘要截断于250字)

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