The roles of Notch3 on the cell proliferation and apoptosis induced by CHIR99021 in NSCLC cell lines: a functional link between Wnt and Notch signaling pathways

PLoS One. 2013 Dec 18;8(12):e84659. doi: 10.1371/journal.pone.0084659. eCollection 2013.

Abstract

Wnt and Notch signaling pathways both play essential roles and interact closely in development and carcinogenesis, but their interaction in non-small-cell lung cancer (NSCLC) is poorly unknown. Here we investigated the effects of CHIR99021, a Wnt signaling agonist, or Notch3-shRNA, or the combined application of CHIR99021 and Notch3-shRNA on cell proliferation and apoptosis, as well as the expressions of Notch3, its downstream genes, cyclinA and caspase-3. Our results showed that CHIR99021 up-regulated the expression of Notch3 protein and HES1 and HEYL mRNA. CHIR99021 promoted cell proliferation and the expression of cyclinA, which were inhibited by Notch3-shRNA in these three cell lines. Moreover, Notch3-shRNA significantly attenuated the positive effects of CHIR99021 on cell proliferation and cyclinA in H460 and H157. As for apoptosis, Notch3-shRNA induced cell apoptosis and increased the expression of caspase-3, whereas CHIR99021 showed the different effects in these three cell lines. The inhibitory effect of CHIR99021 on apoptosis was significantly weakened by Notch3-shRNA only in H460. Overall, although the effects of CHIR99021 and the combined application of CHIR99021 and Notch3-shRNA on the cell proliferation and apoptosis aren't completely similar in the three cell lines, our findings still indicate that Notch3 signaling can be activated by canonical Wnt signaling and a functional link between Wnt and Notch signaling pathways exists in NSCLC, at least, which partially is associated with their regulations on the expressions of cyclinA and caspase-3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Blotting, Western
  • Carcinoma, Non-Small-Cell Lung / metabolism
  • Carcinoma, Non-Small-Cell Lung / physiopathology*
  • Caspase 3 / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects*
  • Cyclin A / metabolism
  • DNA Primers / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression Regulation / drug effects*
  • Humans
  • Microscopy, Fluorescence
  • Pyridines / pharmacology*
  • Pyrimidines / pharmacology*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Real-Time Polymerase Chain Reaction
  • Receptor, Notch3
  • Receptors, Notch / genetics*
  • Receptors, Notch / metabolism
  • Wnt Signaling Pathway / drug effects
  • Wnt Signaling Pathway / physiology*

Substances

  • Chir 99021
  • Cyclin A
  • DNA Primers
  • NOTCH3 protein, human
  • Pyridines
  • Pyrimidines
  • RNA, Small Interfering
  • Receptor, Notch3
  • Receptors, Notch
  • Caspase 3

Grants and funding

The Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (to Chunyan Li, No. 20091001), Liaoning Nature Science Project (to Chunyan Li, No. 201102258). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.