[Role of hydrogen peroxide in promoting proliferation and transformation of rat liver oval cell line WB-F344].

OBJECTIVE

To study the role of hydrogen peroxide (H2O2) in promoting proliferation and transformation of rat liver oval cell line WB-F344.

METHODS

Culturing WB-F344 cells were stimulated directly by H2O2. The effect of H2O2 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-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were both promoted by stimulating continuously with H2O2 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 H2O2 of low concentration for only one time. The normal WB cells and the WB cells initiated with MNNG were transformed by action with H2O2 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 H2O2 were related to the low dose concentration in promoting proliferation and transformation of liver oval cells indicating its important role in hepatocarcinogenesis. Therefore antioxidants should be able to provide a new clue in prevention and cure of hepatoma.