过氧化氢促肝卵圆细胞株WB-F344的增殖及转化作用

Role of Hydrogen Peroxide in Promoting Proliferation and Transformation of Rat Liver Oval Cell Line WB-F344

目的 探讨过氧化氢（H_2O_2）对肝卵圆细胞株WB-F344的促增殖及转化作用。 方法 用H_2O_2直接作用于培养的WB-F344细胞，以MTT 比色试验、3H-TdR掺入液闪计数观察其对细胞的促增殖效应；经N-甲基-N′-硝基-N-亚硝基胍（MNNG）启动的和未经MNNG启动的WB细胞用小剂量H_2O_2连续作用促其转化，以形态学观察、核型分析及甲基纤维素半固体培养进行检测。 结果 小剂量的H_2O_2一次作用具有明显的促进WB细胞增殖的效应，连续作用28 d，可促进经MNNG启动的和未经MNNG启动的WB细胞发生转化，形态上呈现典型的转化细胞的特征；核型分析显示染色体数目改变，染色体结构畸变率明显升高；在甲基纤维素半固体培养基中形成克隆。 结论 小剂量H_2O_2可诱使肝卵圆细胞增殖、转化，提示H_2O_2在肝癌发生中可能具有重要作用，抗氧化作用可能在肝癌的防治中具有一定的意义。

Objective To study the role of hydrogen peroxide（H_2O_2）in promoting proliferation and transfo- rmation of rat liver oval cell line WB-F344． Methods Culturing WB-F344 cells were stimulated directly by H_2O_2． The effect of H_2O_2 in promoting proliferation of WB cells was investigated by using MTT colorimetric analysis and 3H-TdR incorporation liquid scintillation counter． The normal WB cells and the WB cells initiated with N-methy-N′-nitro-N-nitrosoguanidine(MNNG) were both promoted by stimulating continuously with H_2O_2 of low concentration(800 nmol/L)． The transformation effect was tested by morphologic observation, karyotype analysis, and anchorage-independent growth assay． Results Proliferation of WB cells was induced obviously by H_2O_2 of low concentration for only one time． The normal WB cells and the WB cells initiated with MNNG were transformed by action with H_2O_2 of low concentration continuously for 28 days and typical morphologic characters of transformed cells were observed． In karyotype analysis the cells chromosome number changed and the frequency of structure aberration raised dramatically． Otherwise the transformed cells could form clone on self-solid culture medium． Conclusions The biological effects of H_2O_2 were related to the low dose concentration in promoting proliferation and transformation of liver oval cells indicating its important role in hepatocarc- inogenesis． Therefore antioxidants should be able to provide a new clue in prevention and cure of hepatoma．