α5-nAChR modulates nicotine-induced cell migration and invasion in A549 lung cancer cells

Exp Toxicol Pathol. 2015 Sep;67(9):477-82. doi: 10.1016/j.etp.2015.07.001. Epub 2015 Jul 20.

Abstract

Cigarette smoking is the most important risk factor in the development of human lung cancer. Nicotine, the major component in tobacco, not only contributes to carcinogenesis but also promotes tumor metastasis. By binding to nicotinic acetylcholine receptors (nAChRs), nicotine induces the proliferation and migration of non-small cell lung cancer. Recently studies have indicated that α5-nAChR is highly associated with lung cancer risk and nicotine dependence. Nevertheless, it is unclear whether nicotine promotes the migration and invasion through activation of α5-nAChR in lung cancer. In the present study, A549 cell was exposed to 1μN nicotine for 8, 24 or 48h. Wound-healing assay and transwell assay were used to evaluate the capability of A549 cell migration and cell invasion, respectively. Silencing of α5-nAChR was done by siRNA. Western blotting and PCR were used to detect α5-nAChR expression. Nicotine can induce activation of α5-nAChR in association with increased migration and invasion of human lung cancer A549 cell. Treatment of cells with α5-nAChR specific siRNA blocks nicotine-stimulated activation of α5-nAChR and suppresses A549 cell migration and invasion. Reduction of α5-nAChR resulted in upregulation of E-cadherin, consistent with E-cadherin being inhibitive of cancer cell invasion. These findings suggest that nicotine-induced migration and invasion may occur in a mechanism through activation of α5-nAChR, which can contribute to metastasis or development of human lung cancer.

Keywords: A549 cell; E-cadherin; Invasion; Migration; Nicotine; α5-nAChR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cadherins / genetics
  • Cadherins / metabolism
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cell Movement / drug effects*
  • Gene Expression
  • Gene Silencing
  • Humans
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology*
  • Neoplasm Invasiveness
  • Nicotine / toxicity*
  • RNA, Small Interfering / genetics
  • Receptors, Nicotinic / genetics
  • Receptors, Nicotinic / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Cadherins
  • RNA, Small Interfering
  • Receptors, Nicotinic
  • nicotinic receptor alpha5 subunit, human
  • Nicotine