The influence of TNF-α and Ang II on the proliferation, migration and invasion of HepG2 cells by regulating the expression of GRK2

Cancer Chemother Pharmacol. 2017 Apr;79(4):747-758. doi: 10.1007/s00280-017-3267-z. Epub 2017 Mar 18.

Abstract

Purpose: Hepatocellular carcinoma (HCC) is a common digestive system malignancy that is associated with a poor prognosis. This study researched the interaction of tumor necrosis factor-α (TNF-α) and angiotensin II (Ang II) in HCC cells proliferation, migration and invasion and examined their influence on the expression of G protein-coupled receptor kinase 2 (GRK2) and relevant receptors.

Methods: Cell Counting Kit-8 and Transwell assays were performed to evaluate the effects of TNF-α and Ang II on HepG2 cells proliferation, migration and invasion. Flow cytometry was used to investigate the expression of tumor necrosis factor receptor 1 (TNFR1), angiotensin II type 1 (AT1R) and type 2 receptors (AT2R) on the surface of HepG2 cells. Additionally, Western blot was performed to assess the modulation of GRK2 expression by TNF-α and Ang II in HepG2 cells. Meanwhile, GRK2 siRNA-transfected HepG2 cells were used to confirm the effects of GRK2, TNF-α and Ang II on the proliferation, migration and invasion of GRK2-knockdown HCC cells. Finally, the expression of TNF-α, Ang II, TNFR1, AT1R, AT2R and GRK2 proteins in HCC, tumor-adjacent and normal liver tissues were tested by immunohistochemistry.

Results: The data demonstrated that TNF-α and Ang II can enhance the proliferation, migration and invasion of HepG2 cells through suppressing GRK2 expression but that the two reagents combined did not have synergistic effects. Moreover,overexpression of TNFR1 and AT1R perhaps promoted the formation and progression of HCC, while high AT2R expression had the opposite effect.

Conclusions: This study provides new ideas for the prevention and treatment of HCC by researching the interaction and probable mechanism of different bioactive factors associated with HCC.

Keywords: Angiotensin II; GRK2; Hepatocellular carcinoma; Invasion; Proliferation; Tumor necrosis factor-α.

MeSH terms

  • Angiotensin II / pharmacology*
  • Angiotensin II / therapeutic use
  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Agents / therapeutic use
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Cell Movement / drug effects*
  • Cell Proliferation / drug effects*
  • Flow Cytometry
  • G-Protein-Coupled Receptor Kinase 2 / biosynthesis*
  • G-Protein-Coupled Receptor Kinase 2 / genetics
  • Gene Knockdown Techniques
  • Humans
  • Neoplasm Invasiveness / pathology
  • RNA, Small Interfering / genetics
  • Receptor, Angiotensin, Type 1 / biosynthesis
  • Receptor, Angiotensin, Type 2 / biosynthesis
  • Receptors, Tumor Necrosis Factor, Type I / biosynthesis
  • Tumor Necrosis Factor-alpha / pharmacology*
  • Tumor Necrosis Factor-alpha / therapeutic use

Substances

  • Antineoplastic Agents
  • RNA, Small Interfering
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Tumor Necrosis Factor, Type I
  • Tumor Necrosis Factor-alpha
  • Angiotensin II
  • GRK2 protein, human
  • G-Protein-Coupled Receptor Kinase 2