| [Objective] On the basis of the previous work, use ROR?agonist SR1078?and ROR? inhibitors T0901317 act on MGC803 cells, to investigate the effect and mechanism of ROR? on human gastric carcinoma cells EMT and migration, through the DADS with SR1078 contras, reveal whether DADS is a ROR alpha agonist, further prove that DADS is specific for the prevention and treatment of gastric cancer.[Method] Human gastric carcinoma cells MGC803 were divided into untreated group, DADS-treated group, the SR1078-treated group, the T0901317-treated group, the SR1078-and-DADS-treated group, the T0901317-and-DADS- treated group.Inverted microscope observation human gastric carcinoma cells of MGC803 morphology.Immunofluorescence technique and Western blot to detect the ROR-? expression in MGC803 cells, and Wnt/Beta-catenin pathways and molecular ?--catenin, related to EMT Rac1, TGF-? 1 and Vimentin,E-cadherin protein expression of each group. MTT, wound healing and Transwell invasion chamber assay were used respectively to detect proliferation and invasion migration ability of each group.Co-immunoprecipitation were employed to confirm RORa binding and localization with?--catenin in human gastric.[Results]As exposed to inversion microscope and optics microscope,cells treat with T0901317 are long spindle, pleomorphism, cytoplasm out long and slender projections,similar to the control. cells with SR1078,most of cells are round, spindle cells decreased significantly, cytoplasm swelled, atypi ruduce. Cells are gather together that on DADS, turn round, protuberant disappear, atypia ruduce too. Cells treat with SR1078 jion DADS, spindle become less than DADS and SR1078 respectively. DADS join to the T0901317, round cells increased, spindle decrease compare with only treat T0901317. The MTT showed that,exposure of human gastric cancer MGC803 cells to DADS and SR1078(a RORa activator) respectively, proliferation ability was significantly inhibited than without drug treatment, whereas the proliferation ability of cells that with T0901317(a ROR? inhibitor) was promoted than without drug treatment, DADS jiont to T0901317, the proliferation ability was inhibited than T0901317.Wound healing and Transwell invasion chamber assays revealed that, cells treated with DADS and SR1078 respectively, the migration and invasion were significantly decreased comparing with without drug treatment,cells with T0901317,the migration and invasion was increased than the control.Western blot showed that,DADS and SR1078 significantly upregulated the protein expression of ROR? and E-cadherin, but significantly downregulated the protein expression of Rac1,TGF-?1,Vimentin compare with the control(P<0.05),whereas the T0901317 was downregulated the protein expression of ROR? and E-cadherin, significantly upregulated the protein expression of Rac1, TGF-? 1, Vimentin than the control(P<0.05), DADS combined to T0901317 downregulated the protein expression of ROR? and E-cadherin,significantly upregulated the protein expression of Rac1, TGF-? 1,Vimentin,. Compared to the T0901317. DADS and SR1078 significantly downregulated the expression of nuclear ?-catenin and T0901317 upregulated the expression of nuclear ?-catenin(P<0.05),DADS.Immunofluorescence assay indicated that,cell staining of ROR?,E-cadherin in cells with DADS and SR1078 respectively more than that without drug treatment; Rac1, TGF-? 1, Vimentin were less than that without drug treatment,whereas cell staining of ROR?,E-cadherin in cells with T0901317 less than that without drug treatment and Rac1, TGF-? 1,Vimentin were more than that without drug treatment. Co-immunoprecipitation revealed that DADS and SR1078 promote the binding of ?-catenin and ROR?,T0901317 was suppression the binding of ?-catenin and ROR?.[Conclusions] 1. DADS regulated the Wnt/?-catenin pathway through ROR?, and then inhibit the EMT, migration and invasion in MGC803 cells. 2. ROR alpha agonist can strengthen the effect of DADS inhibit the EMT invasion and migration in MGC803 cells.3. DADS have ROR alpha agonist effects. |
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