For Than Hepg2, Tumor Stem Cell Marker Cd133 Molecules, The Influence Of Afp And Its Biological Significance Of The Research

Objective：Investigate impact of telbivudine on hepatoma cell line HepG2cell tumor stemcell markers CD133and alpha-fetoprotein (AFP)， to discover the molecular biologymechanism of nucleoside drugs treatment of patients with hepatocellular carcinoma to providenew experimental data and theoretical basis for the comprehensive treatment of liver cancer.Methods:In this study, HepG2cells for experimental cell,experiment was divided into twogroups:experimental group (telbivudine group) and control group.cell culture to80%coveredwith flasks, the experimental group was given containing cells culture medium for telbivudine(10umol/L), the control group only received the cell culture medium. The next day to replacethe cell culture medium. Continue to cultured cells for9days, the cells were collected.1)1×107cells/ml for flow cytometry double staining of HepG2cell surface expression of CD133:2)1x107cells/ml was used for chemiluminescence immunoassay intracellular AFP levels;3)1×107cells/ml was used for real-time fluorescence quantitative polymerase chain reaction(Real-Time PCR) detection of intracellular expression of CD133and AFP.Results:1．Flow cytometry showed that telbivudine group and control group of CD133positivecells in the rate respectively of1.9%±0.04and0.83%±0.02, Chi-square test for statisticalsignificant difference (P <0.05) between the two groups, suggesting that for telbivudinetreatment of the HepG2cell surface of tumor stem cell markers CD133expression decreased,2．For telbivudine group and control group HepG2cells of CD133mRNA expressionwere0.609±0.057,0.956±0.132, statistical T-test, there are significant differences betweenthe two groups (P <0.05).3．Two groups of cells in6-well culture plates for3days, take the cell lysate usingchemiluminescence immunoassay, measured two groups of cells within the AFP values were:180.32±3.92149.07±3.57, T-test, of AFP expression differences between the two groups inHepG2cells significantly, with statistical significance (P <0.05).4．The two groups of cells cultured for9days, the cells of the same number of cells toobtain total RNA, were taken20ng/ul of RNA used for Real-time PCR detection, telbivudinegroup and control group,AFP expression were1.061±0.034,2.507±1.379, T-test showedstatistically significant differences between the two groups (P <0.05). Conclusion: the intervention of telbivudine in HepG2cells can inhibit cell tumor stem cellmarkers CD133expression, reduced AFP synthesis, this may be the application of nucleosidedrugs treating hepatitis, delaying one of the molecular biological mechanism of hepatocellularcarcinoma.