Bruton's agammaglobulinemia tyrosine kinase (Btk) regulates TPA‑induced breast cancer cell invasion via PLCγ2/PKCβ/NF‑κB/AP‑1‑dependent matrix metalloproteinase‑9 activation

Oncol Rep. 2021 May;45(5):56. doi: 10.3892/or.2021.8007. Epub 2021 Mar 24.

Abstract

Bruton's agammaglobulinemia tyrosine kinase (BTK) is an important cytoplasmic tyrosine kinase involved in B‑lymphocyte development, differentiation, and signaling. Activated protein kinase C (PKC), in turn, induces the activation of mitogen‑activated protein kinase (MAPK) signaling, which promotes cell proliferation, viability, apoptosis, and metastasis. This effect is associated with nuclear factor‑κB (NF‑κB) activation, suggesting an anti‑metastatic effect of BTK inhibitors on MCF‑7 cells that leads to the downregulation of matrix metalloproteinase (MMP)‑9 expression. However, the effect of BTK on breast cancer metastasis is unknown. In this study, the anti‑metastatic activity of BTK inhibitors was examined in MCF‑7 cells focusing on MMP‑9 expression in 12‑O‑tetradecanoylphorbol‑13‑acetate (TPA)‑stimulated MCF‑7 cells. The expression and activity of MMP‑9 in MCF‑7 cells were investigated using quantitative polymerase chain reaction analysis, western blotting, and zymography. Cell invasion and migration were investigated using the Matrigel invasion and cell migration assays. BTK inhibitors [ibrutinib (10 µM), CNX‑774 (10 µM)] significantly attenuated TPA‑induced cell invasion and migration in MCF‑7 cells and inhibited the activation of the phospholipase Cγ2/PKCβ signaling pathways. In addition, small interfering RNA specific for BTK suppressed MMP‑9 expression and cell metastasis. Collectively, results of the present study indicated that BTK suppressed TPA‑induced MMP‑9 expression and cell invasion/migration by activating the MAPK or IκB kinase/NF‑κB/activator protein‑1 pathway. The results clarify the mechanism of action of BTK in cancer cell metastasis by regulating MMP‑9 expression in MCF‑7 cells.

Keywords: Bruton's agammaglobulinemia tyrosine kinase; matrix metalloproteinase‑9; protein kinase C; MCF‑7 cells.

MeSH terms

  • Adenine / analogs & derivatives
  • Adenine / pharmacology
  • Adenine / therapeutic use
  • Agammaglobulinaemia Tyrosine Kinase / antagonists & inhibitors
  • Agammaglobulinaemia Tyrosine Kinase / metabolism*
  • Breast Neoplasms / chemically induced
  • Breast Neoplasms / drug therapy
  • Breast Neoplasms / genetics
  • Breast Neoplasms / pathology*
  • Cell Movement / drug effects
  • Cell Movement / genetics
  • Female
  • Gene Expression Regulation, Neoplastic
  • Humans
  • MAP Kinase Signaling System / drug effects
  • MAP Kinase Signaling System / genetics
  • MCF-7 Cells
  • Matrix Metalloproteinase 9 / genetics*
  • Matrix Metalloproteinase 9 / metabolism
  • NF-kappa B / metabolism
  • Neoplasm Invasiveness / pathology
  • Neoplasm Invasiveness / prevention & control
  • Phospholipase C gamma / metabolism
  • Piperidines / pharmacology
  • Piperidines / therapeutic use
  • Tetradecanoylphorbol Acetate / toxicity
  • Transcription Factor AP-1 / metabolism

Substances

  • NF-kappa B
  • Piperidines
  • Transcription Factor AP-1
  • ibrutinib
  • Agammaglobulinaemia Tyrosine Kinase
  • BTK protein, human
  • PLCG2 protein, human
  • Phospholipase C gamma
  • MMP9 protein, human
  • Matrix Metalloproteinase 9
  • Adenine
  • Tetradecanoylphorbol Acetate

Grants and funding

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean Government (NRF-2015R1D1A1A01057605, 2017R1A5A2015061), Republic of Korea. The funding bodies had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.