The Effect of Sulindac on Wnt Signaling in Hepatocellular Carcinoma Cell Line SMMC7721 Transfected with HBV X Gene
|Keywords||Hepatitis B virus X gene Non-steroidal anti-inflammatory drugs Sulindac Wnt signaling pathway Hepatic carcinoma|
Objective To investigate the non- steroidal anti-inflammatory drug sulindac (Sulindac) of the hepatitis B virus (HBV) X gene transfection the liver cancer SMMC7721 cells Department of the Wnt signaling pathway . With liposomal transfection method HBx eukaryotic expression vector pcDNA3-X transiently transfected the SMMC7721 cells (7721/HBx), to transfer transfected with empty vector pcDNA3 SMMC7721 cell group (7721/pcDNA3) of untransfected plasmid SMMC7721 cell group ( 7721 ) as the control , after transfection 72h by RT-PCR and Western blot detection of HBx expression ; 24 hours after transient transfection and various concentrations of sulindac intervention 7721/HBx group , and 7721/pcDNA3 - group 72 h, Western blot and immune cells chemical detection of β -catenin expression , Western blot was used to detect the expression of cyclinD1 . The results of RT-PCR and Western blot , 7721/HBx group in RNA and protein levels were HBx expression , 7721/pcDNA3 group and 7721 group no expression . Compared with 7721/pcDNA3 group and the 7721 group , Western blot shows 7721/HBx group of β - catenin , cyclinD1 expression levels , immunocytochemistry shows 7721/HBx group of β-catenin expression levels deep nuclear staining 7721/pcDNA3 group and 7721 group no significant difference between the Western blot and immunocytochemistry shows a certain concentration of sulindac 48h after the intervention 7721/HBx group , and 7721/pcDNA3 - cells can down-regulate the expression of β-catenin and cyclinD1 down to 7721/HBx group than 7721/pcDNA3 group was significantly . Conclusion HBx gene successful transient transfection into SMMC7721 cells , activation of the Wnt signaling pathway , and at the same time raised cyclinD1 expression . Sulindac can inhibit liver cancer cells by β-catenin cut Wnt signaling pathway down cyclinD1 expression and HBx positive HCC cells sensitive sulindac .