Knockdown of RAD54B expression reduces cell proliferation and induces apoptosis in lung cancer cells

J Int Med Res. 2019 Nov;47(11):5650-5659. doi: 10.1177/0300060519869423. Epub 2019 Sep 26.

Abstract

Objective: RAD54 homolog B (RAD54B), a member of the SNF2/SWI2 superfamily, is implicated in homologous recombination, and high RAD54B expression predicts the prognostic outcomes of lung adenocarcinoma. However, its role in lung carcinogenesis was unclear so this was determined in the present study.

Methods: We evaluated the gene and protein expression of RAD54B in 15 lung adenocarcinoma tissues and matched adjacent healthy lung tissues by real-time PCR, immunohistochemical staining, and western blotting. A549 lung cancer cells were transduced with lentivirus carrying small hairpin RNA (shRNA) against RAD54B (shRAD54B) or control shRNA (shCtrl), and cell proliferation, viability, apoptosis, and caspase 3/7 activity were evaluated.

Results: RAD54B protein expression was significantly higher in lung adenocarcinoma tissues than in healthy lung tissues. RAD54B gene expression was high in A549 cells but was efficiently knocked down using shRAD54B with an infection efficiency of 80% and a knockdown ratio of 72.2% compared with shCtrl. Suppressing RAD54B expression in A549 cells significantly reduced cell proliferation and caspase 3/7 activity, and significantly increased the apoptotic rate.

Conclusions: RAD54B exerts an oncogenic role in lung cancer cell proliferation.

Keywords: DNA damage response; Knockdown; RAD54B; apoptosis; lung cancer; oncogenesis; proliferation.

MeSH terms

  • A549 Cells
  • Apoptosis* / genetics
  • Cell Proliferation
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Gene Expression Regulation, Neoplastic
  • Gene Knockdown Techniques*
  • Humans
  • Lung Neoplasms / genetics
  • Lung Neoplasms / pathology*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • RNA, Small Interfering / metabolism

Substances

  • Nuclear Proteins
  • RAD54B protein, human
  • RNA, Small Interfering
  • DNA Helicases