Stimulatory heterotrimeric GTP-binding protein augments cisplatin-induced apoptosis by upregulating Bak expression in human lung cancer cells

Cancer Sci. 2009 Jun;100(6):1069-74. doi: 10.1111/j.1349-7006.2009.01136.x. Epub 2009 Mar 23.

Abstract

The present study aimed to investigate the effect of the stimulatory heterotrimeric GTP-binding (Gs) protein signaling system on cisplatin-induced apoptosis of lung cancer cells and its underlying mechanism as an attempt to develop a novel strategy to improve the therapeutic efficacy of cisplatin. Overexpression of the constitutively active alpha subunit of Gs (GalphasQL) in A549 human lung cancer cells increased cisplatin-induced apoptosis, and knockdown of Galphas with small hairpin RNA decreased the percentage of apoptotic cells. GalphasQL increased the expression of the proapoptotic proteins B-cell leukemia/lymphoma-2 genes (Bcl-2) homologous antagonist killer protein (Bak) and Bcl-2 associated X protein (Bax), and decreased the expression of the antiapoptotic proteins Bcl-2 and Bcl-Xlong protein. Knockdown of Bak blocked the augmentative effects of GalphasQL. GalphasQL decreased the degradation rate of the Bak protein, and increased Bak mRNA transcript levels. GalphasQL increased Bak-luciferase activity in a protein kinase A and cyclic AMP response element-dependent manner. GalphasQL also augmented cisplatin-induced apoptosis of H1299 human lung cancer cells that lack functional p53. From this study, it is concluded that Galphas augments cisplatin-induced apoptosis of lung cancer cells partially through upregulating Bak expression by increasing transcription and by decreasing the rate of protein degradation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Carcinoma, Non-Small-Cell Lung / genetics
  • Carcinoma, Non-Small-Cell Lung / metabolism
  • Carcinoma, Non-Small-Cell Lung / pathology
  • Cell Line, Tumor
  • Cisplatin / pharmacology*
  • Cyclic AMP Response Element-Binding Protein / metabolism
  • DNA Primers
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic
  • Genes, Reporter
  • Heterotrimeric GTP-Binding Proteins / metabolism*
  • Humans
  • Luciferases / genetics
  • Lung Neoplasms / genetics
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology*
  • Transfection
  • bcl-2 Homologous Antagonist-Killer Protein / genetics*

Substances

  • BAK1 protein, human
  • CREB1 protein, human
  • Cyclic AMP Response Element-Binding Protein
  • DNA Primers
  • bcl-2 Homologous Antagonist-Killer Protein
  • Luciferases
  • Heterotrimeric GTP-Binding Proteins
  • Cisplatin