| The hepatocellular carcinoma (HCC) plays an important part in the incidence and mortality of human malignant tumors. Many studies have shown that chemical factors are related to hepatocarcinogenesis. However the mechanisms by which the carcinogens cause the disease are unclear. It has been suggested that Reactive Oxygen Species (ROS) might be involved in hepatocarcinogenesis, but no definitive evidences, especially in the study of cellular and molecular level in vitro are available to prove the view.The liver oval cell line WB-F344 derives from adult normal Fisher 344 male rats. It's a kind of liver infantine epithelial cells and has characteristics of liver stem cells. Many studies in vitro and in vivo imply that it has the role as precursor cells of hepatocacinoma, which indicates its function in hepatocarcinogenesis. Tsao et al have observed that MNNG acted several times could induce WB cells transformation, but the study of inducing its malignant transformation directly by ROS has not been reported.In present, for understanding the relationship between ROS and hepatocarcinogenesis, we have studied on the role and mechanisms of H2O2 in promoting WB cell proliferation and transformation.Part I the role of H2O2 in promoting proliferation and transformation of WB cells. 1. Using different experimental methods that indicate cellular proliferation, we obtained the correlative and corresponding results. When H2O2 of low concentration (800nmol/L) stimulate the quiescent WB cell, the 570nm fluorescent absorption rise by MTT colorimetric analysis, which indicate the increase of the number of cells. The H2O2 also induced the increase in 3H-Tdr incorporation and DNA synthesis by means of the measurement in a liquid scintillation counter. Also the H2O2 of low concentration can cause an increase in the mitoses of WB cell by morphologic examination of Giemsa staining. The experiments above demonstrate that H2O2 of low concentration can evidently promote the proliferation of WB cells. Moreover, the proliferation induced by H2O2 could... |
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