The Preliminary Research Of The Effect And Mechanism Of Pokemon On The Expression Of TGF-β1 In Gasric Cancer Cells

Objective:To explore the preliminary research of the effect and mechanism of pokemon on the expression of TGF-β1 in gasric cancer cells,and to analyze its clinicopathologic correlationMethods: the experiments includ two parts. : The first part :The MGC-803 gastric cancer cell lines in vitro culture was the research object, through transfercted the PcDNA3.1-Pokemon and PcDNA3.1 vector into MGC-803 cells,the expression of Pokemon was detected by Q-PCR and Western blot,then the expression of TGF-β1 was detected by fluorescent microscopy,the expression of TGF-β1mRNA was detected by Q-PCR, the Part of EMT related protein expression was detected by Western blot; 2) the SGC-7901 gastric cancer cell lines in vitro culture was the research object, through transfercted the PcDNA3. 1-Pokemon and PcDNA3.1 vector into SGC-7901 cells,the expression of Pokemon was detected by Q-PCR and Western blot,then the expression of TGF-β1 was detected by fluorescent microscopy,the expression of TGF-β1mRNA was detected by Q-PCR, the Part of EMT related protein expression was detected by Western blot;The second part:Application of Dual-Luciferase Reporter Assay determinated the expression plasmid of TGF- β1 promoter in MGC- 803 and SGC- 7901 gastric cancer cell line after the transfection of Pokemon。Results:1)After PcDNA3.1-Pokemon were transfected into MGC- 803 and SGC- 7901 gastric cancer cells, the protein and mRNA levels of Pokemon was increased;2)The TGF-β1mRNA levels was decreased significantly by transfected with PcDNA3.1-Pokemon( P<0.05);3) Immunofluorescence microscopy revealed lower TGF-β1 levels after transfected with PcDNA3.1-Pokemon(P<0.05);4)The expression of E-cadherin and ZO-1 protein was increased,and the expression of Vimentin and Slug protein was decreased(P<0.05);5)Higher Pokemon treatment led to decreased the luciferase activity of TGF-β1 by the luciferase reporter assay(P<0.05)。Conclusion:1)Pokemon regulated EMT process by inhibiting the expression of TGF-β1 in MGC-803 and SGC-7901 gastric cancer cell lines;2)Pokemon inhibit the expression of TGF-β1 in gastric cancer cells by inhibiting TGF-β1 promoter activity.