4.5 Article

Visualization of the cancer cell cycle by tissue-clearing technology using the Fucci reporter system

期刊

CANCER SCIENCE
卷 112, 期 9, 页码 3796-3809

出版社

WILEY
DOI: 10.1111/cas.15034

关键词

antitumor drug; cell cycle; Fucci system; metastasis; tissue-clearing technology

类别

资金

  1. Ministry of Education, Culture, Sports, Science, and Technology of Japan KAKENHI [17H06326]
  2. Japan Society for the Promotion of Science KAKENHI [19K16604, 20K16212, 15H05774, 20H00513]
  3. Grants-in-Aid for Scientific Research [19K16604, 20K16212, 20H00513] Funding Source: KAKEN

向作者/读者索取更多资源

Tissue-clearing technology combined with the Fucci reporter system visualized the cell cycles of primary and metastatic tumors, showing different fluorescence patterns of the Fucci reporter in tumor colonies among various organs and even within the same organs. The Fucci reporter cells were also used to evaluate the effects of anti-tumor drugs on cancer cells.
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 G(2)/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.

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