The changes in secretory phenotypes of senescent human gastric epithelial cells

Objectives To investigate the effects of cell aging on the disorders relating to gastric mucosa aging. Methods A treatment of 200 μmol/L H2O2 was used to induce senescence of human gastric epithelial cell line GES-1, and the cell growth curve was monitored.Senescence secretory phenotypes were observed by detecting the protein level of p53 and p16INK4a with senescence-associated β-galactosidase(SA-β-gal)staining and Western blot testing.The mRNA levels of senescence-associated secretory phenotype(SASP)factors in human gastric epithelial GES-1 cell including IL-1β, IL-6, IL-8, TGF-β、IFN-γ, and VEGF-A were detected by RT-PCR.The mRNA expression levels of IL-1β, IL-6, IL-8, TGF-β, IFN-γ, and VEGF in the conditioned medium were detected by ELISA analysis. Results The 200 μmol/L H2O2-induced GES-1 cells stopped proliferating after 3 days of treatment, and cells enlarged and flattened at 10 days.The increased SA-β-gal staining(P<0.001)and the increased expression levels of p53 and p16INK4a proteins indicated the success of establishing the aging model of GES-1.The mRNA levels of IL-1β, IL6, IL8, TGF-β, and IFNγ were higher(t=2.94, 3.38, 3.15, 3.64, 2.97; P=0.015, 0.000, 0.000, 0.000, 0.000)and the mRNA level of VEGF-A was lower(t=2.31, P=0.20)in senescent GES-1 cells than in the control group.In the conditioned medium of senescent GES-1 cells, the levels of IL-1β, IL6, IL8, TGF-β1, and IFNγ were higher in the H2O2-induced group[(3.12±0.21)μg/L, (4.26±0.15)μg/L, (3.37±0.14)μg/L, (5.34±0.19)μg/L, and(2.90±0.47)μg/L]than in the negative control group[(0.24±0.04, 0.04±0.07, 0.52±0.02, 1.05±0.10, 0.52±0.02, respectively, P<0.001)], while the level of VEGF was lower in the H2O2-induced group than in the negative control group(0.21±0.03)μg/Lvs(0.59±0.07)μg/L(P<0.05). Conclusions The changes in senescence-associated secretory phenotype factors of the aging human gastric epithelial cells induced by oxidative stress may promote chronic gastritis and gastric cancer. Key words: Gastric mucosa; Epithelial cells; Cell aging; Senescence-associated secretory phenotype