The Mechanism On Perfluorodecanoic Acid (PFDA) Promoting Gastric Epithelial Cells Proliferation

OBJECTIVE To research the influences of PFDA, PFOA,PFOS on the proliferation of gastric epithelial cell, and the molecular mechanisms of PFDA affecting the proliferation of gastric epithelial cell. Environmental pollutants have a great promotion on the incidence of gastric cancer.Research could help to elucidate the contribution of environmental Pollutants to gastric cancer development. Make the community realize the hazardness of environmental Pollution.METHODS1.To detect the effecth of PFDA, PFOA,PFOS on the proliferation of gastric epithelial cell,Gastric epithelial cells were cultured and treated with PFDA for 72 h and selected 300 cells for Clonogenic assay.Counting the number of clones after the clones were visible.2.After treating Gastric epUhelial cells with PFDA for 72 hours, we detected the effects of PFDA on the apoptosis, cell cycles with Flow Cytometry,protein level of autophagy-related proteins of P62,LC3 using Western Blotting. Detecting the effects of senescence in this molecular mechanisms of PFDA promoting Gastric epithelial cells proliferation with β-Galactosidase staining.3.Microarray analysis the possthle target genes which promote the cell Proliferation and further verify via qRT-PCR.4.The effects of TCF4,PLA2G2 A on the molecular mechanisms of PFDA promoting the proliferation of gastric epithelial cell.Gastric epithelial cells were treated with PFDA for 24 hour’s,the TCF4,PLA2G2 A expression plasmid were transfected for 48 hour’s, we tested the expression of TCF4,PLA2G2 A with qRT-PCR,and th detect the senescence changes using p-Galactosidase staining.RESULTS1.Colony formation result showed that PFOA,PFOS,PFDA promok the proliferation of Gastric epithelial cells(AGS),but the capability of promoting the proliferation is different, the order is PFDA> PFOS>PFOA.2.Flow Cytometry result showed PFD A has no effect on the cell cycles, but the apoptosis of Gastric epithelial cells was inhibited,mainly the early stage of apoptosis. The expression of autophagy-related proteins P62,LC3 increase after treated with PFDA,which was not obvious,suggesting that autophagy plays a certain role in inhibiting the proliferation of Gastric epithelial cells.3.Microarray analysis displayed that PFDA inhibits the expression of TCF4,PLA2G2 A mRNA significantly.4.PFDA negatively regulates PLA2G2 A through TCF4 th inhthit the senescence of Gastric epithelial cells,and the number of senescet cells increase significantly after TCF4,PLA2G2 A expression plasmid transfecthd.Conclusion1.PFDA promotes the proliferation of gastric epithelial cell.2.PFDA promotes the proliferation of gastric epithelial cell through inhthitingth e senescence.3.PFDA negatively regulates PLA2G2A through TCF4 to inhthit the senescence of Gastric epithelial cells,which playes an important role in the molecular mechanisms of PFDA promoting the proliferation of gastric epithelial.