Effects of SAM level on proliferation of human liver cancer cell line HepG2 under the treatment of HGF

Abstract

Objective To investigate the effects of SAM level on proliferation of human liver cancer cell line HepG2 under the treatment of hepatocyte growth factor (HGF).Methods We established cell systems of different levels of SAM by transfecting pGCSIL-GFP-MAT2A siRNA plasmids into HepG2 cells or adding C3-Ado.The cell system was tested by RP-HPLC.The cell growth curve was drawn by methylthiazol tetrazolium (MTT) assay.Flow cytometry (FCM) was adopted to analyze cell cycle and apoptosis.Results After pGCSIL-GFP-MAT2AsiRNA was transfected successfully,the SAM level was increased to ( 1.35 ± 0.06 ) nmol/mg protein.MTT assay revealed that SAM could suppress the growth of HepG2 cells.Floy cytometry showed that the number of cells in G0/G1 phase was increased by 23.42% and that in G2/M phase decreased by 40.61%,and HepG2 cell apoptosis rate was 34.58%; After C3-Ado being added,the SAM level in HepG2 cells was decreased to (0.37 ± 0.03 ) nmol/mg protein,and proliferation of HepG2 cells was accelerated,the number of cells in G0/G1 phase was decreased by 7.73% and that in G2/M phase was increased by 18.18%,and HepG2 cell apoptosis rate was 5.17%.Conclusion The SAM level in HepG2 cells was the key in the process of HGF promoting proliferation.Only the concentration of SAM was lower level,HGF could promote proliferation of HepG2 cells. Key words: Carcinoma,hepatocellular; siRNA; SAM; Cell proliferation