Gap junctions contribute to anchorage-independent clustering of breast cancer cells

BMC Cancer. 2018 Feb 27;18(1):221. doi: 10.1186/s12885-018-4148-5.


Background: Cancer cell aggregation is a key process involved in the formation of clusters of circulating tumor cells. We previously reported that cell-cell adhesion proteins, such as E-cadherin, and desmosomal proteins are involved in cell aggregation to form clusters independently of cell migration or matrix adhesion. Here, we investigated the involvement of gap junction intercellular communication (GJIC) during anchorage-independent clustering of MCF7 breast adenocarcinoma cells.

Methods: We used live cell image acquisition and analysis to monitor the kinetics of MCF7 cell clustering in the presence/absence of GJIC pharmacological inhibitors and to screen a LOPAC® bioactive compound library. We also used a calcein transfer assay and flow cytometry to evaluate GJIC involvement in cancer cell clustering.

Results: We first demonstrated that functional GJIC are established in the early phase of cancer cell aggregation. We then showed that pharmacological inhibition of GJIC using tonabersat and meclofenamate delayed MCF7 cell clustering and reduced calcein transfer. We also found that brefeldin A, an inhibitor of vesicular trafficking, which we identified by screening a small compound library, and latrunculin A, an actin cytoskeleton-disrupting agent, both impaired MCF7 cell clustering and calcein transfer.

Conclusions: Our results demonstrate that GJIC are involved from the earliest stages of anchorage-independent cancer cell aggregation. They also give insights into the regulatory mechanisms that could modulate the formation of clusters of circulating tumor cells.

Keywords: Anchorage-independent aggregation; Cancer cell clustering; Gap junction intercellular communication.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / metabolism
  • Adenocarcinoma / physiopathology*
  • Antigens, CD
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / physiopathology*
  • Cadherins
  • Cell Adhesion
  • Cell Communication*
  • Cell Movement
  • Female
  • Gap Junctions*
  • Humans
  • MCF-7 Cells


  • Antigens, CD
  • CDH1 protein, human
  • Cadherins