Visualization of the cancer cell cycle by tissue-clearing technology using the Fucci reporter system.
Visualization of the cancer cell cycle by tissue-clearing technology using the Fucci reporter system.
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DOI：
10.1111/cas.15034
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发表时间：
2021-09
期刊：
影响因子：
5.7
通讯作者：
Miyazono K
中科院分区：
文献类型：
--
作者：
Takahashi K;Tanabe R;Ehata S;Kubota SI;Morishita Y;Ueda HR;Miyazono K
组织清除技术是一种新兴的成像技术，目前不仅用于神经科学研究，也用于癌症研究。在我们以前的报告中，组织清除方法被用于检测转移性肿瘤。在这里，我们展示了原发瘤和转移瘤的细胞周期是通过使用报告系统的组织清除方法可视化的。首先，我们利用广泛使用的癌细胞株A549和4T1建立了稳定表达基于荧光泛素化的细胞周期指示剂(Fucci)报告的癌细胞系。在小鼠不同的肿瘤接种模型中，研究了Fucci报告基因的荧光模式。有趣的是，肿瘤菌落的Fucci报告基因的荧光模式在不同的器官之间是不同的，甚至在相同器官的菌落之间也不同。用这些Fucci报告细胞也评估了抗肿瘤药物的效果。在所研究的三种抗癌药物中，5-氟尿嘧啶对4T1-Fucci细胞的处理在体内外均通过诱导G2/M期阻滞而产生独特的荧光模式。因此，结合组织清除法和Fucci报告基因有助于分析肿瘤转移和耐药的机制。用4T1-Fucci细胞监测体内Fucci模式。A、B、3D和2D图像显示4T1细胞原位接种于乳房脂肪垫内。实验性肺转移中4T1-Fucci细胞的C、3D和2D图像
Tissue‐clearing technology is an emerging imaging technique currently utilized not only in neuroscience research but also in cancer research. In our previous reports, tissue‐clearing methods were used for the detection of metastatic tumors. Here, we showed that the cell cycles of primary and metastatic tumors were visualized by tissue‐clearing methods using a reporter system. First, we established cancer cell lines stably expressing fluorescent ubiquitination‐based cell cycle indicator (Fucci) reporter with widely used cancer cell lines A549 and 4T1. Fluorescence patterns of the Fucci reporter were investigated in various tumor inoculation models in mice. Interestingly, fluorescence patterns of the Fucci reporter of tumor colonies were different between various organs, and even among colonies in the same organs. The effects of antitumor drugs were also evaluated using these Fucci reporter cells. Of the three antitumor drugs studied, 5‐fluorouracil treatment on 4T1‐Fucci cells resulted in characteristic fluorescent patterns by the induction of G2/M arrest both in vitro and in vivo. Thus, the combination of a tissue‐clearing method with the Fucci reporter is useful for analyzing the mechanisms of cancer metastasis and drug resistance. Monitoring the Fucci patterns in vivo with 4T1‐Fucci cells. A, B, 3D and 2D images of 4T1 cells inoculated orthotopically into the mammary fat‐pad. C, 3D and 2D images of 4T1‐Fucci cells in experimental lung metastasis.
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影响因子：
5.7
作者：
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