Effects Of Moniliformin On Human Hepatoblastoma Cells Biological Behavior And Involved Primary Mechanisms

Objectives:Human hepatoblastoma cell line Hep G2 and Huh7 were used in this subject to explore the effects of moniliformin(MON) on human hepatoblastoma cells biological behavior, and the involved primary mechanisms were explored by proteomic analysis.Methods:1. Effects of MON on human hepatoblastoma cells biological behavior:(1) Effect of MON on cell proliferation was tested by CCK-8.(2) Effect of MON on migration was tested by transwell assay and the wound healing assay.(3) Effect of MON on oncogenicity was tested by soft agar assay.2. Primary mechanisms that MON affected human hepatoblastoma cells biological behavior by proteomic analysis:(1) Sample preparation and quantity of 2-DE.(2) IEF and SDS-PAGE.(3) Scanning and analysis of 2-DE gels.(4) Sample preparation of mass spectroscopy and identification by MALDI-TOF-MS.Results:1. Effect of MON on biological behavior of human hepatoblastoma cells:(1) After exposure at 0.001 ?g/ml-1 ?g/ml of MON over four days, there was no influence on the cell proliferation of Hep G2 and Huh7 cells. The cell proliferation of Hep G2 was decreased when the cells were exposed at 0.5 ?g/ml and 1 ?g/ml of MON after five days(P?0.001); the cell proliferation of Huh7 was retarded when the cells were exposed at 1 ?g/ml of MON after five days(P?0.01).(2) The migration ability of Hep G2 and Huh7 was enhanced when the cells were treated with 0.001 ?g/ml-0.5 ?g/ml of MON: the migration ability of Hep G2 was significantly promoted at 0.1 ?g/ml of MON(P?0.001); the migration ability of Huh7 was remarkablely improved at 0.2 ?g/ml of MON(P?0.001).(3) After dealing with 0.001 ?g/ml-0.5 ?g/ml of MON, the colony formation of Hep G2 and Huh7 in soft agar was enhanced: the malignancy of Hep G2 was significantly promoted at 0.1 ?g/ml of MON; the malignancy of Huh7 was remarkablely improved at 0.2 ?g/ml of MON.2 ? Primary mechanisms that MON affected human hepatoblastoma cells biological behavior by proteomic analysis:(1) 1021 protein spots matched successfully between the control groups and the experimental groups, and 8 protein spots which were expressed differently at least1.5 fold between the control groups and the experimental groups, and down-regulated expression in the experimental groups screened and analysed totally by Imaging Master 2D Platinum 7.0.(2) There were 8 protein spots identified by MALDI-TOF-MS.Conclusions:1. In our study, MON could promote migration and malignancy, but could not affect cells proliferation of human hepatoblastoma cells.2. MON could affect the protein expression profiles of hepatocellular carcinoma cells. 8protein spots were identified which were expressed differently at least 1.5 fold between the control groups and the experimental groups, and down-regulated expression in the experimental groups.