The Effect Of Sulindac On Wnt Signaling In Hepatocellular Carcinoma Cell Line SMMC7721 Transfected With HBV X Gene

Aim: To explore the effect of Non-steroidal anti-inflammatory drugs Sulindac on Wnt signaling in hepatocellular carcinoma cell line SMMC7721 transfected with HBV X gene.Methods: The HBV X gene eukaryon expression vector pcDNA3-X was transiently transfected into SMMC7721 cell by lipid-media transfection(7721/HBx) . Untransfected SMMC7721 (7721) and SMMC7721 transfected with pcDNA3(7721/pcDNA3) were used as control.The expression of HBx in SMMC7721 was identified by RT-PCR and Western blot analysis 72h after the transient transfection.7721/HBx and 7721/pcDNA3 cells were treated with Sulindac in different concentration for 48h.The changes of expressions ofβ-catenin were examined by Western blot and immunocytochemistry analysis,with the changes of expression of cyclinD1 by Western blot analysis.Results: RT-PCR and Western-blot analysis showed that 7721/HBx cells transiently express HBV X protein. Western blot and immunocytochemistry analysis showed the protein expression ofβ-catenin and cyclinD1 was much more in 7721/HBx before Sulindac treatment compared with 7721/pcDNA3 and 7721.Western blot and immunocytochemistry analysis showed Sulindac in some concentration could inhibit the protein expression ofβ-catenin and cyclinD1,which were more remarkable in 7721/HBx cells than in 7721/pcDNA3 cells.Conclusions: HBx could up-regulate the expression and regulate the etopic expression ofβ-catenin in hepatocellular carcinoma cell SMMC7721,which lead to the activation of Wnt signaling,while it could also up-regulate cyclinD1.Sulindac could down-regulateβ-catenin and cyclinD1,and inhibit Wnt signaling in HBx positive HCC cells,which were more remarkable than in HBx negative HCC cells.