Effects Of Akt-YAP Signaling Pathway On Glutamine Deprivation Induced Apoptosis In Hela Cells

Objective:To investigate the effects of glutamine starvation on cell growth and viability, and to explore the mechanism of Akt-YAP signaling pathway in the process of glutamine on apoptosis.Methods:Hela cells cultured in vitro were used in all experiments. Hela cells were cultured with different concentrations of FBS and Gin (10%FBS+4mM Gin,10%FBS+Gin-Free, FBS-Free+4mM Gin, FBS-Free+Gln-Free) for24and48hours, cell proliferation was measured using cell counting method. Hela cells were cultured in DMEM medium with different concentrations of Gin (4,2,1,0.5,0.1,0mM) for24hours, cell viability was measured using the Cell Counting Kit, and the protein expression of Bax was measured by Western blot.Hela cells were cultured in DMEM medium with different concentrations of Gin (4,2,1,0.5,0.1,0mM) or with Gin4mM, Gin0mM and Akt inhibitor LY29400240μM+Gln4mM for24hours, and the expression of pAkt and Bax protein was measured by Western blot.Hela cells were cultured in DMEM medium with different concentrations of Gin (4,2,1,0.5,0.1,0mM) for24hours, and the expression of pYAP and YAP protein was measured by Western blot; Using siRNA to suppress YAP expression, normal cells and transfected cells were cultured in DMEM medium with Gin4mM or0mM for24hours, Bax protein expression was measured. Hela cells were cultured in DMEM medium with Gln4mM, Gln0mM or Akt inhibitor LY29400240μM+Gln4mM for24hours, the expression of pYAP and YAP protein in the cytoplasm and nucleus was measured by Western blot. Results:Glutamine starvation reduced the growth rate and the number of Hela cells, whether adding FBS or not (P<0.05); Cell viability decreased with the reduction of Gin concentrations (P<0.05), and Bax expression increased;With the reduction of glutamine concentration pAkt expression decreased, both glutamine starvation and Akt inhibitor increased the expression Bax protein;With the reduction of Gin concentration pYAP expression decreased; Bax expression increased in Gin starvation of normal cells, while it reduced in YAP siRNA transfected cells; Both glutamine starvation and Akt inhibitor decreased pYAP expression, and increased YAP expression in the nucleus.Conclusion:Gin starvation inhibited cell growth and viability, induced apoptosis of Hela cells; Akt-YAP signaling pathway involved in apoptosis induced by Gin starvation in Hela cells.