细胞内稳态S-腺苷甲硫氨酸浓度对人肝细胞生长因子促肝癌细胞增殖效应中的影响  

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

作　　者：王刚[1] 何跃明[1] 刘权焰[1] 刘志苏[1] 

机构地区：[1]武汉大学中南医院肝胆外科,430071

出　　处：《中华实验外科杂志》2010年第12期1832-1834,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30872491/C160402）;湖北省自然科学基金资助项目（2009CDB292）;湖北省卫生厅基金资助项目（nx200718）

摘　　要：目的 观察细胞内稳态S-腺苷甲硫氨酸（SAM）水平的变化对人肝细胞生长因子（HGF）促肝癌细胞增殖效应的影响.方法 采用pGCSIL-GFP-MAT2AsiRNA重组质粒转染HepG2细胞升高细胞内SAM浓度,SAM竞争抑制剂C3-Ado降低SAM浓度,用RP-HPLC验证不同SAM浓度的细胞体系,并用噻唑蓝（MTT）比色法、流式细胞术（FCM）检测各细胞体系中的HGF促增殖效应.结果 HGF处理的HepG2细胞转染重组质粒后细胞内SAM浓度升高至（1.35±0.06）nmol/mg蛋白质,细胞增殖活性降低,G0/G1期细胞增加23.42%,G2/M期细胞减少40.61%,细胞凋亡率34.58%;当加入C3-Ado,细胞内SAM浓度下降至（0.37±0.03）nmol/mg蛋白质,细胞增殖活性升高,G0/G1期细胞减少7.73%,G2/M期细胞增加18.18%,细胞凋亡率5.17%.结论 HGF在肝癌细胞中发挥促有丝分裂效应的必要条件是肝癌细胞内稳态SAM浓度处于较低水平.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.

关 键 词：癌 肝细胞 RNA干扰 S-腺苷甲硫氨酸 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]