| Objective:To study the effect of over expression Trop2 on migration of the human gastric mucosal epithelial cell and preliminary analyses the mechanism.Methods:We explored the mRNA and protein expression level of Trop2 in GES-1 cell line by Real-time RT-PCR and Western-blot;constructed OE-Trop2 and VE-Trop2 plasmid and transfected into GES-1 cell line and detected the efficiency of the transfection;cell wound healing assay and transwell migration assay explored the change of the migration in GES-1 after transfected OE-Trop2 plasmid;Western-blot explored the change of the EMT phenomenon in GES-1 cell line after over expression Trop2.Results:Compared with gastric cancer cell line BGC823 and MGC803,the mRNA and protein expression level of Trop2 is lower;over expression Trop2 plasmid transfected in GES-1 and the mRNA expression level of Trop2 in OE-Trop2 group was 6.18±0.52 times higher than that of the control group,whereas no difference were detected between control group and VE-Trop2 group,meanwhile,the protein expression level was similar to that of the mRNA expression level;after transfected plasmid 72h,the migration rate of GES-1 was 50%in VE-Trop2 group and 95%in OE-Trop2 group by cell wound healing assay(P<0.05);after transfected OE-Trop2 plasmid 24h,the number of migration cell was 41±6 in VE-Trop2 group,whereas 87±6 in OE-Trop2 group by transwell migration assay(P<0.05);the expression level of E-cadherin in GES-1 was down-regulated,meanwhile,fibronectin and vimentin were up-regulated after transfected OE-Trop2 by western-blot.Conclusion:Trop2 could promoted migration and deterioration of GES-1 cell line through induced EMT phenomenon. |
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