Physiological Hypoxia (Physioxia) Impairs the Early Adhesion of Single Lymphoma Cell to Marrow Stromal Cell and Extracellular Matrix. Optical Tweezers Study

Int J Mol Sci. 2018 Jun 26;19(7):1880. doi: 10.3390/ijms19071880.

Abstract

Adhesion is critical for the maintenance of cellular structures as well as intercellular communication, and its dysfunction occurs prevalently during cancer progression. Recently, a growing number of studies indicated the ability of oxygen to regulate adhesion molecules expression, however, the influence of physiological hypoxia (physioxia) on cell adhesion remains elusive. Thus, here we aimed: (i) to develop an optical tweezers based assay to precisely evaluate single diffuse large B-cell lymphoma (DLBCL) cell adhesion to neighbor cells (mesenchymal stromal cells) and extracellular matrix (Matrigel) under normoxia and physioxia; and, (ii) to explore the role of integrins in adhesion of single lymphoma cell. We identified the pronouncedly reduced adhesive properties of lymphoma cell lines and primary lymphocytes B under physioxia to both stromal cells and Matrigel. Corresponding effects were shown in bulk adhesion assays. Then we emphasized that impaired β1, β2 integrins, and cadherin-2 expression, studied by confocal microscopy, account for reduction in lymphocyte adhesion in physioxia. Additionally, the blockade studies conducted with anti-integrin antibodies have revealed the critical role of integrins in lymphoma adhesion. To summarize, the presented approach allows for precise confirmation of the changes in single cell adhesion properties provoked by physiological hypoxia. Thus, our findings reveal an unprecedented role of using physiologically relevant oxygen conditioning and single cell adhesion approaches when investigating tumor adhesion in vitro.

Keywords: cell-to-cell interactions; diffuse large B-cell lymphoma (DLBCL); integrins; mesenchymal stromal cells (MSC); optical tweezers; physiological hypoxia (physioxia); single cell adhesion.

MeSH terms

  • Antigens, CD / metabolism
  • Bone Marrow / pathology*
  • Cadherins / metabolism
  • Cell Adhesion
  • Cell Adhesion Molecules / metabolism
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Survival
  • Collagen / metabolism
  • Drug Combinations
  • Extracellular Matrix / metabolism*
  • Humans
  • Hypoxia / pathology*
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Integrin beta1 / metabolism
  • Laminin / metabolism
  • Lasers
  • Lymphoma, Large B-Cell, Diffuse / metabolism
  • Lymphoma, Large B-Cell, Diffuse / pathology*
  • Mesenchymal Stem Cells / metabolism
  • Optical Tweezers*
  • Proteoglycans / metabolism
  • Single-Cell Analysis
  • Stromal Cells / pathology
  • Time Factors

Substances

  • Antigens, CD
  • CDH2 protein, human
  • Cadherins
  • Cell Adhesion Molecules
  • Drug Combinations
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Integrin beta1
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen