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ATP生物发光测定法和流式细胞术在预测子宫癌细胞系放射敏感性中的应用:放射毒性与细胞周期动力学的相关性

The use of ATP bioluminescence assay and flow cytometry in predicting radiosensitivity of uterine cancer cell lines: correlation of radiotoxicity and cell cycle kinetics.

作者信息

Nguyen H N, Sevin B U, Averette H E, Gottlieb C F, Perras J, Ramos R, Donato D, Penalver M

机构信息

Department of Obstetrics & Gynecology, University of Miami School of Medicine, Florida 33101.

出版信息

Gynecol Oncol. 1992 Jul;46(1):88-96. doi: 10.1016/0090-8258(92)90202-t.

Abstract

Radiotherapy remains an important part of uterine cancer treatment. This study was designed to evaluate the potential of the ATP bioluminescence assay and flow cytometry for predicting radiosensitivity. Correlation of these two modalities revealed important insights into the relationship of radiotoxicity and cell kinetic effects. Six human uterine cancer cell lines were used: AE7, ECC1, HEC1A, HEC1B, AN3, and SKUT1B. Doses of cobalt 60 were 0, 1, 2, 5, 8, and 10 Gy. The ATP bioluminescence assays were performed on Day 7. Cell samples were taken at 0, 24, 48, 72, 96, and 168 hr for flow cytometry. The linear-quadratic model was used to fit survival data and mean inactivation dose D was calculated. Among parameters such as D, alpha and beta coefficients, and surviving fraction at 2 Gy (SF2), both D and SF2 correlated best with survival data. Radiation effects on the cell cycle did not correlate with D and revealed two distinct patterns: either a G1 accumulation with mild G2 block or a G1 depletion and severe G2 block. The S cells consistently demonstrated a biphasic pattern with an initial reduction followed by an accumulation. In summary, the ATP assay was shown to have potential in the study of radiosensitivity. Radiation-induced cell kinetics appeared to vary with intrinsic cellular differences and, thus, could not be used to predict radiosensitivity.

摘要

放射治疗仍然是子宫癌治疗的重要组成部分。本研究旨在评估ATP生物发光测定法和流式细胞术预测放射敏感性的潜力。这两种方法的相关性揭示了对放射毒性与细胞动力学效应关系的重要见解。使用了六种人子宫癌细胞系:AE7、ECC1、HEC1A、HEC1B、AN3和SKUT1B。钴60的剂量分别为0、1、2、5、8和10 Gy。在第7天进行ATP生物发光测定。在0、24、48、72、96和168小时采集细胞样本用于流式细胞术检测。采用线性二次模型拟合生存数据并计算平均灭活剂量D。在诸如D、α和β系数以及2 Gy时的存活分数(SF2)等参数中,D和SF2与生存数据的相关性最佳。辐射对细胞周期的影响与D不相关,并呈现出两种不同的模式:要么是G1期积累并伴有轻度G2期阻滞,要么是G1期减少并伴有严重G2期阻滞。S期细胞始终呈现出双相模式,先是减少,随后积累。总之,ATP测定法在放射敏感性研究中显示出潜力。辐射诱导的细胞动力学似乎因细胞内在差异而异,因此不能用于预测放射敏感性。

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