Research On The Mechanism Of Esophagus Squamous Cancer Cell Proliferation And Invasion By Prostaglandin E₂ Via Activating EP₂/EGFR Signaling Pathway

Objective: To investigate the effect of prostaglandin E₂?PGE₂?and its receptor subtype EP₂ on the expression of epidermal growth factor receptor?EGFR?by in vitro cell culture experiments,and then regulate the proliferation and invasion of esophageal cancer cells,which can promote the secretion of a variety of cytokines through this pathway revealing the inflammatory factors in the development of esophageal cancer in the pathogenesis of esophageal cancer for the treatment of new targets to provide a basic theoretical basis.Methods: 1.In vitro cell culture of ESCC cell lines EC109 and TE-1 and normal esophageal epithelial cells;2.The levels of EP₂,EGFR m RNA in EC109 and TE-1 cells were detected by real-time quantitative PCR?RT-PCR?with normal esophageal epithelial cells as negative control group;The expression of EP₂,EGFR and phosphorylated EGFR?p EGFR?protein in TE-1 cells were detected by Western Blot?WB?.3.The expression of EP₂,EGFR and p EGFR protein were observed by RT-PCR and WB in the treatment of EC109 and TE-1 cells treated with PGE₂ and EP₂-specific agonists Butaprost and EP₂ inhibitors.The cell proliferation rate was detected by WST-8?CCK-8?method.Transwell invasion assay was used to detect the cell invasion ability.4.EC109 and TE-1 cells were treated with PGE₂ and EP₂-specific agonists Butaprost and EP₂ inhibitors respectively.The levels of matrix metalloproteinase-9?MMP-9?,vascular endothelium Growth factor?VEGF?expression level;5.The difference between the two groups was statistically significant by using the statistical method,the comparison between the two groups,and the comparison between the two groups before and after the intervention.Results: 1.Real-time quantitative PCR?RT-PCR?showed that the expression of EP₂,EGFR m RNA in EC109 and TE-1 cells were significantly higher than those in control group?P <0.05?.2.The expression of EP₂,EGFR and p EGFR protein in EC109 and TE-1 cells were significantly higher than those in control group?P <0.05?by Western Blot?WB?.3.RT-PCR showed the expression of EP₂ m RNA was decreased and the difference was statistically significant?P <0.05?,and EGFR m RNA unchanged?P> 0.05?in the cells treated with PGE₂;The expression of EP₂ m RNA was significantly increased?P <0.05?and the difference was statistically significant?P <0.05?,EGFR m RNA unchanged?P>0.05?in the cells treated with Butaprost;The expression of EP₂ m RNA was decreased and he difference was statistically significant?P <0.05?,EGFR m RNA unchanged?P>0.05?in the cells treated with EP₂ inhibitors.4.The expression of EP₂ and p EGFR protein was significantly higher than that of the control group?P <0.05?,and EGFR protein was unchanged?P> 0.05?in the cells treated with PGE₂;The expression of EP₂ and p EGFR protein was increased?P <0.05?,EGFR protein was unchanged?P> 0.05?in the cells treated with Butaprost;EP₂ and p EGFR protein expression were decreased?P <0.05?,EGFR protein was unchanged?P> 0.05?in the cells treated with EP₂ inhibitors.5.The expression of EC109 and TE-1 cells was detected by PGE₂ and Butaprost,and the cell proliferation rate was increased by CCK-8 method?P <0.05?.Transwell invasion test increased the invasive ability?P <0.05?.By EP₂ inhibitors intervention,proliferation and invasion were decreased?P <0.05?,the difference was statistically significant.6.The expression of MMP-9 and VEGF in the EC109 and TE-1 cells was significantly higher than that in the control group by PGE₂ and Butaprost?P <0.05?.The levels of MMP-9 and VEGF were significantly decreased by EP₂ inhibitors?P <0.05?.7.There was no significant difference in the expression of EC109 and TE-1 between the observation group and the control group?P> 0.05?.Conclusion: The expression of EP₂ in PGE₂ receptor subtypes,EGFR and p EGFR in esophageal squamous cell carcinoma cells can be detected by in vitro cell culture.PGE₂ can promote the secretion of MMP-9 and VEGF by EP₂ activation of p EGFR pathway,which enhance directly or indirectly the proliferation and invasion of esophageal squamous cell carcinoma cell lines,affecting the biological behavior of esophageal squamous cell carcinoma.