Investigation Of PAFc-Interacting Proteins In Hepatocelluar Carcinoma Cells

Objective: Identify PAFc-interacting proteins in Hepatocelluar carcinoma(HCC)cells,to provide the underlying mechanisms of PAFc’s function in HCC.Methods:Co-immunoprecipitation method was employed to purify PAFc interacting protein from Hepatocelluar carcinoma cells.The purified proteins were loaded on SDS-polyacrylamide gel electrophoresis(SDS-PAGE),and then coomassie blue stain was utilized to display the protein bands which were cut back and subjected into in-gel digestion,subsequently for mass spectrometric analysis.The proteins of interest were selected and high efficient overexpressed in 293 T cells.Immunoprecipitation method and Western blot experiments were employed to verify the interactions.Results: 1.PAFc interacting proteins were identified and screened by Co-immunoprecipitation method and mass spectrometric analysis.Fli1 and SALL4 were chosen as interested potential PAFc interacting proteins,by bioinformatics analysis.2.Fli1 and SALL4 were respectively overexpressed,in the 293 T cells.3.SALL4 was further verified to be PAFc interacting protein by CO-IP and WB.Conclusions: We identifed SALL4 to be PAFc interacting proteins.These results provide new evidence for further understanding of PAFc function and molecular mechanisms of HCC.The interaction of PAFc with SALL4 in Hepatocelluar carcinoma cells may be related to occurrence and development of HCC.