Post-transcriptional repression of circadian component CLOCK regulates cancer-stemness in murine breast cancer cells

Abstract

Disruption of the circadian clock machinery in cancer cells is implicated in tumor malignancy. Studies on cancer therapy reveal the presence of heterogeneous cells, including breast cancer stem-like cells (BCSCs), in breast tumors. BCSCs are often characterized by high aldehyde dehydrogenase (ALDH) activity, associated with the malignancy of cancers. In this study, we demonstrated the negative regulation of ALDH activity by the major circadian component CLOCK in murine breast cancer 4T1 cells. The expression of CLOCK was repressed in high-ALDH-activity 4T1, and enhancement of CLOCK expression abrogated their stemness properties, such as tumorigenicity and invasive potential. Furthermore, reduced expression of CLOCK in high-ALDH-activity 4T1 was post-transcriptionally regulated by microRNA: miR-182. Knockout of miR-182 restored the expression of CLOCK, resulted in preventing tumor growth. Our findings suggest that increased expression of CLOCK in BCSCs by targeting post-transcriptional regulation overcame stemness-related malignancy and may be a novel strategy for breast cancer treatments.

Data availability

The full data of microarray analysis have been deposited in National Center for Biotechnology Information gene expression omnibus (miRNA microarray, Accession#:GSE157655; mRNA microarray, Accession#:GSE103598). All data generated or analysed during this study are included in the manuscript and supporting files. Source data files of the quantitative data have been provided for all figures.

The following previously published data sets were used

Article and author information

Author details

  1. Takashi Ogino

    Pharmaceutics, Kyushu University, Higashi-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
  2. Naoya Matsunaga

    Glocal Healthcare Science, Kyushu University, Higashi-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
  3. Takahiro Tanaka

    Pharmaceutics, Kyushu University, Higashi-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
  4. Tomohito Tanihara

    Pharmaceutics, Kyushu University, Higashi-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
  5. Hideki Terajima

    Department of Biological Sciences, The University of Tokyo, Tokyo, Japan
    Competing interests
    The authors declare that no competing interests exist.
  6. Hikari Yoshitane

    Department of Biological Sciences, The University of Tokyo, Tokyo, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6319-3354
  7. Yoshitaka Fukada

    Department of Biological Sciences, The University of Tokyo, Tokyo, Japan
    Competing interests
    The authors declare that no competing interests exist.
  8. Akito Tsuruta

    Pharmaceutics, Kyushu University, Higashi-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
  9. Satoru Koyanagi

    Glocal Healthcare Science, Kyushu University, Higashi-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
  10. Shigehiro Ohdo

    Pharmaceutics, Kyushu University, Higashi-ku, Japan
    For correspondence
    ohdo@phar.kyushu-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4795-9764

Funding

Ministry of Education, Culture, Sports, Science and Technology (Grant-in-Aid for Scientific Research A,16H02636)

  • Shigehiro Ohdo

Japan Agency for Medical Research and Development (JP20am0101091)

  • Shigehiro Ohdo

Japan Agency for Medical Research and Development (JP21am0101091)

  • Shigehiro Ohdo

Ministry of Education, Culture, Sports, Science and Technology (Challenging Exploratory Research,17H06262)

  • Shigehiro Ohdo

Ministry of Education, Culture, Sports, Science and Technology (Challenging Exploratory Research,20K21484)

  • Satoru Koyanagi

Ministry of Education, Culture, Sports, Science and Technology (Challenging Exploratory Research,20K21901)

  • Naoya Matsunaga

Ministry of Education, Culture, Sports, Science and Technology (Scientific Research B,18H03192)

  • Naoya Matsunaga

Ministry of Education, Culture, Sports, Science and Technology (Specially Promoted Research,17H06096)

  • Yoshitaka Fukada

Ministry of Education, Culture, Sports, Science and Technology (Scientific Research B,25440041)

  • Hikari Yoshitane

Japan Agency for Medical Research and Development (PRIME,17937210)

  • Hikari Yoshitane

Ministry of Education, Culture, Sports, Science and Technology (JSPS KAKENHI Grant,17J01969)

  • Takashi Ogino

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Caigang Liu, Shengjing Hospital of China Medical University, China

Ethics

Animal experimentation: All experimental procedures were performed after approval and following the guidelines of Kyushu University (approval number: A20-131-0).

Version history

  1. Received: December 30, 2020
  2. Accepted: April 22, 2021
  3. Accepted Manuscript published: April 23, 2021 (version 1)
  4. Version of Record published: May 6, 2021 (version 2)

Copyright

© 2021, Ogino et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,955
    views
  • 256
    downloads
  • 11
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Takashi Ogino
  2. Naoya Matsunaga
  3. Takahiro Tanaka
  4. Tomohito Tanihara
  5. Hideki Terajima
  6. Hikari Yoshitane
  7. Yoshitaka Fukada
  8. Akito Tsuruta
  9. Satoru Koyanagi
  10. Shigehiro Ohdo
(2021)
Post-transcriptional repression of circadian component CLOCK regulates cancer-stemness in murine breast cancer cells
eLife 10:e66155.
https://doi.org/10.7554/eLife.66155

Share this article

https://doi.org/10.7554/eLife.66155

Further reading

    1. Cancer Biology
    2. Cell Biology
    Julian J A Hoving, Elizabeth Harford-Wright ... Alison C Lloyd
    Research Article

    Collective cell migration is fundamental for the development of organisms and in the adult, for tissue regeneration and in pathological conditions such as cancer. Migration as a coherent group requires the maintenance of cell-cell interactions, while contact inhibition of locomotion (CIL), a local repulsive force, can propel the group forward. Here we show that the cell-cell interaction molecule, N-cadherin, regulates both adhesion and repulsion processes during rat Schwann cell (SC) collective migration, which is required for peripheral nerve regeneration. However, distinct from its role in cell-cell adhesion, the repulsion process is independent of N-cadherin trans-homodimerisation and the associated adherens junction complex. Rather, the extracellular domain of N-cadherin is required to present the repulsive Slit2/Slit3 signal at the cell-surface. Inhibiting Slit2/Slit3 signalling inhibits CIL and subsequently collective Schwann cell migration, resulting in adherent, nonmigratory cell clusters. Moreover, analysis of ex vivo explants from mice following sciatic nerve injury showed that inhibition of Slit2 decreased Schwann cell collective migration and increased clustering of Schwann cells within the nerve bridge. These findings provide insight into how opposing signals can mediate collective cell migration and how CIL pathways are promising targets for inhibiting pathological cell migration.

    1. Cancer Biology
    2. Structural Biology and Molecular Biophysics
    Johannes Paladini, Annalena Maier ... Stephan Grzesiek
    Research Article

    Abelson tyrosine kinase (Abl) is regulated by the arrangement of its regulatory core, consisting sequentially of the SH3, SH2, and kinase (KD) domains, where an assembled or disassembled core corresponds to low or high kinase activity, respectively. It was recently established that binding of type II ATP site inhibitors, such as imatinib, generates a force from the KD N-lobe onto the SH3 domain and in consequence disassembles the core. Here, we demonstrate that the C-terminal αI-helix exerts an additional force toward the SH2 domain, which correlates both with kinase activity and type II inhibitor-induced disassembly. The αI-helix mutation E528K, which is responsible for the ABL1 malformation syndrome, strongly activates Abl by breaking a salt bridge with the KD C-lobe and thereby increasing the force onto the SH2 domain. In contrast, the allosteric inhibitor asciminib strongly reduces Abl’s activity by fixating the αI-helix and reducing the force onto the SH2 domain. These observations are explained by a simple mechanical model of Abl activation involving forces from the KD N-lobe and the αI-helix onto the KD/SH2SH3 interface.