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用于动态监测电离辐射对人肿瘤细胞系和原代成纤维细胞影响的新型实时细胞分析平台。

Novel real-time cell analysis platform for the dynamic monitoring of ionizing radiation effects on human tumor cell lines and primary fibroblasts.

作者信息

Mán Imola, Szebeni Gábor J, Plangár Imola, Szabó Emilia R, Tőkés Tünde, Szabó Zoltán, Nagy Zoltán, Fekete Gábor, Fajka-Boja Roberta, Puskás László G, Hideghéty Katalin, Hackler László

机构信息

Avidin Ltd., Szeged H‑6726, Hungary.

Department of Neurology, Faculty of Medicine, University of Szeged, Szeged H‑6725, Hungary.

出版信息

Mol Med Rep. 2015 Sep;12(3):4610-1619. doi: 10.3892/mmr.2015.4004. Epub 2015 Jun 26.

Abstract

Translational research in radiation oncology is important for the detection of adverse radiation effects, cellular responses, and radiation modifications, and may help to improve the outcome of radiation therapy in patients with cancer. The present study aimed to optimize and validate a real‑time label‑free assay for the dynamic monitoring of cellular responses to ionizing radiation. The xCELLigence system is an impedance‑based platform that provides continuous information on alterations in cell size, shape, adhesion, proliferation, and survival. In the present study, various malignant human primary fibroblast cells (U251, GBM2, MCF7, A549, HT‑29) were exposed to 0, 5 and 10 Gy of Cobalt60 radiation. As well as the xCELLigence system, cell survival and proliferation was evaluated using the following conventional end‑point cell‑based methods: Clonogenic, MTS, and lactate dehydrogenase assays, and apoptosis was detected by fluorescence‑activated cell sorting. The effects of ionizing radiation were detected for each cell line using impedance monitoring. The real‑time data correlated with the colony forming assay results. At low cell densities (1,000‑2,000 cells/well) the impedance‑based method was more accurate at monitoring dose‑dependent changes in the malignant human primary fibroblast cell lines, as compared with the end‑point assays. The results of the present study demonstrated that the xCELLigence system may be a reliable and rapid diagnostic method for the monitoring of dynamic cell behavior following radiation. In addition, the xCELLigence system may be used to investigate the cellular mechanisms underlying the radiation response, as well as the time‑dependent effects of radiation on cell proliferation and viability.

摘要

放射肿瘤学中的转化研究对于检测辐射不良反应、细胞反应和辐射修饰很重要,并且可能有助于改善癌症患者的放射治疗效果。本研究旨在优化和验证一种用于动态监测细胞对电离辐射反应的实时无标记检测方法。xCELLigence系统是一个基于阻抗的平台,可提供有关细胞大小、形状、粘附、增殖和存活变化的连续信息。在本研究中,将各种恶性人原代成纤维细胞(U251、GBM2、MCF7、A549、HT-29)暴露于0、5和10 Gy的钴60辐射。除了xCELLigence系统外,还使用以下传统的基于细胞终点的方法评估细胞存活和增殖:克隆形成、MTS和乳酸脱氢酶测定,并通过荧光激活细胞分选检测细胞凋亡。使用阻抗监测检测每种细胞系的电离辐射效应。实时数据与集落形成试验结果相关。在低细胞密度(1000-2000个细胞/孔)下,与终点试验相比,基于阻抗的方法在监测恶性人原代成纤维细胞系中剂量依赖性变化方面更准确。本研究结果表明,xCELLigence系统可能是一种可靠且快速的诊断方法,用于监测辐射后动态细胞行为。此外,xCELLigence系统可用于研究辐射反应背后的细胞机制,以及辐射对细胞增殖和活力的时间依赖性影响。

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