Visualizing the effect of tumor microenvironments on radiation-induced cell kinetics in multicellular spheroids consisting of HeLa cells
Visualizing the effect of tumor microenvironments on radiation-induced cell kinetics in multicellular spheroids consisting of HeLa cells
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DOI：
10.1016/j.bbrc.2013.08.093
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发表时间：
2013-10-04
影响因子：
3.1
通讯作者：
Miura, Masahiko
中科院分区：
文献类型：
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作者：
Kaida, Atsushi;Miura, Masahiko
在这项研究中，我们可视化了肿瘤微环境对辐射诱导的肿瘤细胞动力学的影响。为此，我们利用了直径约为 500 μm 的多细胞球体模型，由表达基于荧光泛素化的细胞周期指示剂 (Fucci) 的 HeLa 细胞组成。在活体球体中，共焦激光扫描显微镜使我们能够清楚地监测深度达 60 μm 的细胞动力学。令人惊讶的是，相对于单层培养的细胞，在球体的外部区域观察到 G2 停滞的显着延长。 Scale 是一种使组织光学透明的水性试剂，可以更深入地观察球体内部。照射后约 16 小时，红色荧光细胞部分（可能是静止的 G0 细胞部分）变得与由增殖细胞组成的外部部分不同，其中大多数显示出表明 G2 停滞的绿色荧光。此后，红细胞部分开始发出绿色荧光并保持长时间的 G2 停滞。因此，我们第一次可视化了球体中辐射诱导的 G2 停滞的延长以及外部和内部部分之间的细胞动力学差异。 (c) 2013 Elsevier Inc. 保留所有权利。
In this study, we visualized the effect of tumor microenvironments on radiation-induced tumor cell kinetics. For this purpose, we utilized a multicellular spheroid model, with a diameter of similar to 500 mu m, consisting of HeLa cells expressing the fluorescent ubiquitination-based cell-cycle indicator (Fucci). In live spheroids, a confocal laser scanning microscope allowed us to clearly monitor cell kinetics at depths of up to 60 mu m. Surprisingly, a remarkable prolongation of G2 arrest was observed in the outer region of the spheroid relative to monolayer-cultured cells. Scale, an aqueous reagent that renders tissues optically transparent, allowed visualization deeper inside spheroids. About 16 h after irradiation, a red fluorescent cell fraction, presumably a quiescent G0 cell fraction, became distinct from the outer fraction consisting of proliferating cells, most of which exhibited green fluorescence indicative of G2 arrest. Thereafter, the red cell fraction began to emit green fluorescence and remained in prolonged G2 arrest. Thus, for the first time, we visualized the prolongation of radiation-induced G2 arrest in spheroids and the differences in cell kinetics between the outer and inner fractions. (c) 2013 Elsevier Inc. All rights reserved.