Objective: To study the role of superoxide anion (O2.-) in promoting proliferation and transformation of rat liver oval cell strain WB-F344.
Methods: WB-F344 cells cultured were stimulated directly by O2.- generated by interaction of xanthine with xanthine oxidase (X-XO). The effect of O2.- in promoting proliferation of WB cells was investigated by using MTT colorimetric analysis, 3H-Tdr incorporation liquid scintillation counter and 3H-Tdr incorporation autoradiography. WB cells initiated with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) were promoted by stimulating continuously with O2.- of low concentration (X: 100 mumol/L, XO: 0.2 mU/ml). The transformation effect was tested by morphologic observation, karyotype analysis and anchorage-independent growth assay.
Results: Proliferation of WB cells was induced obviously by O2.- of low concentration for only one time. WB cells initiated with MNNG were transformed by action with O2.- of low concentration continuously for 15 d and typical morphologic character of transformed cells was observed. In karyotype analysis the cells chromosome number changed and the frequency of structure aberration raised dramatically. Also the transformed cells could form clone on self-solid culture medium.
Conclusions: The biological effect of O2.- was related closely with its dose; The effects of low concentration in promoting proliferation and transformation of liver oval cells indicate its important role in hepatocarcinogenesis and antioxidation was able to provide a new clue in prevention and cure of hepatoma.