Cytotoxicity Differences Of Troglitazone On HepG2 And CD133 Liver Cancer Stem Cells

Objective Isolation and identification CD133 liver cancer stem cells in HepG2 cells,to investigate the cytotoxicity of troglitazone on HepG2 and its liver stem cells.Methods the CD133-positive and negative population in the HepG2 cell line were sorted by flow cytometry;The cells of CD133 sphere formation,colony formation assay and Transwll invasion migration assay,BALB/c nude mice in vivo tumor formation are used to identify proliferative capacity;liver cancer stem cells drug resisitance was evaluated by MTT;The toxicity of Tro to CD133 liver cancer stem cells was detected by MTT assay,Automatic biochemical analyzer was used to detect the contents of AST,LDH,TP,ALB and BUN changes in cell supernatants after administration,The effect of Tro on the total activity of CYP450 and the level of ROS was examined by fluorescence method to compare the cytotoxicity differences.Flow cytometry analysis of Tro cycle after cell cycle and apoptosis changes to study Tro showed cytotoxicity differencesResults CD133 expression in liver cancer stem cells were found to be(0.72±0.05)% in HepG2 and 98.7% in CD133+ cells;CD133,c-Myc and Oct4 protein in CD133+ cells were higher than those in parental cells;Sphere formation,colony formation assay and Transwll invasion migration assay of CD133+cells(P<0.05,P<0.01),compared with HepG2 group,the ability of CD133+ cell tumor formation was significantly increased(P<0.01).At the same time,CD133+ cells were mostly in G0/G1 phase,G2/M phase was not blocked,and the drug resistance of sorafenlb was higher(P <0.01);Tro treatment 12 h,24 h,48 h,the IC50 of liver cancer stem cells was(150.52 ± 1.25)?mol/L,(99.08±1.90)?mol/L,(43.96±0.71)?mol/L and(14.81±1.30)?mol/L respectively,which was significantly lower than that of HepG2(P<0.01),with a significant dose-effect relationship;TP,LDH,ALB and BUN in liver cancer stem cells treated with 80 ?mol/L Tro for 48 h increased significantly,Wherein the elevated LDH levels higher than parental cells(P <0.05);CYP450 total activity(P <0.05)and ROS levels(P <0.05)significant inhibition,CYP 1A2 degree of inhibition increases with concentration.In the apoptosis,Tro could induce the early apoptosis and necrosis of CD133+ cells,which was significantly higher than that of the parent cells,showing the cytotoxicity of the larger Tro(P <0.01).In the cell cycle,(P <0.01),and increased the S phase and G1 / M phase,showing the difference of cell selection toxicity(P <0.01).Conclusion Successful screening and identification of highly proliferative CD133+HepG2 tumor stem cells are more sensitive than HepG2 cells in showing liver cytotoxicity,and show significant cytotoxicity difference,which provides a new idea for the study of CD133+ HepG2 cells specific toxicity.