Interleukin-8-Induced Invasion Assay in Triple-Negative Breast Cancer Cells

Methods Mol Biol. 2020:2108:107-115. doi: 10.1007/978-1-0716-0247-8_9.

Abstract

The pro-inflammatory and pro-angiogenic chemokine interleukin-8 (IL-8, CXCL8) induces proliferation and invasion of solid tumor cells. In many types of solid cancer, including triple-negative breast cancer (TNBC), the IL-8 expression is induced by proteasome inhibition. In this chapter, we describe a protocol for the analysis of TNBC cell invasion induced by IL-8 in response to proteasome inhibition by bortezomib (BZ). Using this approach, we show that BZ increases the invasion ability of TNBC cells, and that the BZ-increased TNBC cell invasion is suppressed by IκB kinase (IKK) inhibition, which also decreases the IL-8 expression. The experimental protocol includes the cell invasion assay, microscopic evaluation of the invading cells, and quantitative analysis of the obtained images. This protocol should be applicable also for measurement of chemokine-induced tumor cell invasion in other types of cancer cells.

Keywords: Bortezomib; Cancer cell invasion; Chemokines; Interleukin-8; IκB kinase; Proteasome inhibition; Triple-negative breast cancer.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers
  • Cell Line, Tumor
  • Cell Movement* / drug effects
  • Cell Proliferation
  • Cytokines / metabolism
  • Data Analysis
  • Female
  • Humans
  • I-kappa B Kinase / metabolism
  • Interleukin-8 / metabolism*
  • Interleukin-8 / pharmacology
  • Molecular Imaging
  • Proteasome Inhibitors / pharmacology
  • Software
  • Triple Negative Breast Neoplasms / metabolism*
  • Triple Negative Breast Neoplasms / pathology

Substances

  • Biomarkers
  • Cytokines
  • Interleukin-8
  • Proteasome Inhibitors
  • I-kappa B Kinase