RRC ID 81407
Author Takahashi K, Tanabe R, Ehata S, Kubota SI, Morishita Y, Ueda HR, Miyazono K.
Title Visualization of the cancer cell cycle by tissue-clearing technology using the Fucci reporter system.
Journal Cancer Sci
Abstract 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.
Volume 112(9)
Pages 3796-3809
Published 2021-9-1
DOI 10.1111/cas.15034
PMID 34145937
PMC PMC8409402
MeSH A549 Cells Adenocarcinoma of Lung / pathology* Animals Antimetabolites, Antineoplastic / administration & dosage Breast Neoplasms / pathology* Cell Cycle* / drug effects Cell Cycle* / genetics Cell Proliferation / drug effects Female Fluorouracil / administration & dosage Genes, Reporter Genetic Vectors / genetics Humans Luminescent Measurements / methods* Luminescent Proteins / genetics Lung Neoplasms / pathology* Mice Mice, Inbred BALB C Mice, Nude Microscopy, Fluorescence / methods Red Fluorescent Protein Transfection Ubiquitination Xenograft Model Antitumor Assays
Resource
DNA material pCAG-HIVgp (RDB04394) pCMV-VSV-G-RSV-Rev (RDB04393)