Verification On The Interaction Between Trefoil Factor 1 And Aldo-keto Reductase A1C1

OBJECTIVE:To analyze the interaction of TFF1 with AKR1C1 and observe the expression and cellular localization of TFF1 and AKR1C1.METHODS:The cDNA fragments encoding TFF1 and AKR1C1 was amplified by polymerase chain reaction(PCR) method,and subsequently were cloned into pCMV-AD and pCMV-BD vector and sequenced,respectively.Plasmids pCMV-NFkB-TFF1 and pCMV-GAL4-AKR1C1 were constructed and then the studies of binding in vivo were carded out by co-immunoprecipitation after co-transfection of the constructs into 293FT cells.The expression and subcellular localization of the fusion protein in the transfected 293-FT cells were analyzed by Western blotting and fluorescence confocal microscopy,respectively.RESULTS:Western blotting confirmed the expression of fusion protein in the transfected 293FT cells.The interactions between TFF1 and interactive proteins AKR1C1 was confirmed by co-immunoprecipitation technique.Fluorescence confocal microscopy showed that the fusion proteins had partial cellular co-localization in 293FT cell.CONCLUSION:This research indicates that AKR1C1 protein may be a newly identified protein which interacts with TFF1.TFF1 has comprehensive roles which remain to be elucidated.Nevertheless,our work offers an opportunity to probe into the role of TFF1 in the pathogenesis of gastrointestinal diseases and Cancer.