超氧阴离子自由基促肝卵圆细胞株WB—F344增殖,转化作用 …

目的 探讨超氧阴离子自由基对肝卵圆细胞株ＷＢ－Ｆ３４４的促增殖及转化作用。方法 用黄嘌呤－黄嘌呤氧化酶（Ｘ－ＸＯ）系统产生的Ｏ２＾－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,＾３Ｈ－Ｔｄｒ掺入放射自显影观察其对细胞的促增殖效应,。细胞经Ｎ－甲基－Ｎ‘－硝基－Ｎ－亚硝基胍（ＭＮＮＧ）启动后,用小剂量的Ｏ２＾－连续作用促其转化,以形态学观察,核型分析及甲基纤维素

Study on the role of superoxide anion in promoting proliferation and transformation of rat liver oval cell line WB-E344

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.