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目的研究氯化汞(HgCl2)对人肝细胞HL-7702细胞增殖、DNA损伤、细胞周期及金属硫蛋白(MT)表达的影响。方法采用四甲基偶氮噻唑蓝法(MTT)、单细胞凝胶电泳(SCGE)、流式细胞术(FCM)及免疫细胞化学方法进行检测。结果HgCl2对HL-7702细胞具有细胞毒性,能够明显抑制HL-7702细胞增殖,并有剂量依赖关系(P<0.05)。HgCl2对HL-7702细胞的抑制率随染毒时间延长而增高。HgCl2可诱导HL-7702细胞产生DNA损伤,随着HgCl2浓度增大和染毒时间延长,DNA损伤率升高(P<0.05)。HgCl2可影响HL-7702细胞DNA合成过程和细胞周期进程。HgCl2染毒导致HL-7702细胞出现G0/G1期阻滞,进入S期的细胞数量减少(P<0.05)。HgCl2可以诱导保护蛋白MT的表达。随HgCl2浓度增大,MT表达较对照组明显增加(P<0.05)。结论HgCl2能够对HL-7702细胞产生细胞毒作用,降低含巯基酶活性、损伤DNA、影响细胞周期进程。
Objective To study the effects of HgCl2 on proliferation, DNA damage, cell cycle and expression of metallothionein (MT) in human hepatocellular HL-7702 cells. Methods MTT assay, single cell gel electrophoresis (SCGE), flow cytometry (FCM) and immunocytochemistry were used for detection. Results HgCl2 was cytotoxic to HL-7702 cells and could inhibit the proliferation of HL-7702 cells in a dose-dependent manner (P <0.05). The inhibitory rate of HgCl2 on HL-7702 cells increased with the prolongation of exposure time. HgCl2 induced DNA damage in HL-7702 cells. With the increase of HgCl2 concentration and exposure time, DNA damage rate increased (P <0.05). HgCl2 can affect DNA synthesis and cell cycle progression in HL-7702 cells. HgCl2 induced HL-7702 cells in G0 / G1 phase arrest, the number of cells into the S phase decreased (P <0.05). HgCl2 can induce the expression of protective protein MT. With the increase of HgCl2 concentration, MT expression was significantly increased (P <0.05). Conclusion HgCl2 can produce cytotoxic effect on HL-7702 cells, reduce the activity of thiol-containing enzyme, damage DNA, and affect the cell cycle progression.