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目的研究微囊藻毒素-LR(MC-LR)对人肝癌HT17细胞周期、凋亡的影响及其可能的作用机制。方法调整HT17细胞密度为1×104/孔,分别设终浓度为0(溶剂对照,0.1%DMSO)、0.05、0.1、0.2、0.4、0.8、2.0、5.0μmol/L的MC-LR染毒组和去铁敏(DFO,1 mmol/L)染毒组以及MC-LR(0.8μmol/L)+DFO(1 mmol/L)染毒组培养24 h,采用四甲基偶氮噻唑蓝(MTT)法检测细胞生长情况。调整HT17细胞密度为1×105/孔,分别设终浓度为0(溶剂对照)、MC-LR(0.8μmol/L)染毒组和DFO(1 mmol/L)染毒组以及DFO(1 mmol/L)+MC-LR(0.8μmol/L)染毒组培养24、48 h,采用流式细胞术检测细胞周期和细胞凋亡情况。结果高剂量(≥0.8μmol/L)MC-LR染毒24 h后细胞存活率明显低于溶剂对照组(P<0.01),DFO+MC-LR染毒组细胞存活率高于0.8μmol/L MC-LR染毒组(P<0.05)。与溶剂对照组相比,MC-LR染毒24、48 h后G0/G1期细胞所占比例和细胞凋亡率均增高,S期细胞所占比例下降(P<0.05或P<0.01);DFO+MC-LR染毒组与MC-LR染毒组染毒24 h后各期细胞所占比例无差异(P>0.05)。DFO+MC-LR染毒组染毒24 h后细胞凋亡率低于MC-LR染毒组(P<0.01)。结论 MC-LR可导致HT17细胞在G0/G1期发生阻滞及细胞凋亡,可能与其诱发的氧化应激、蛋白质磷酸化状态紊乱有关。
Objective To study the effects of microcystin-LR (MC-LR) on the cell cycle and apoptosis of human hepatoma HT17 cells and its possible mechanism. Methods The cell density of HT17 cells was adjusted to 1 × 10 4 / well. MC-LR cells were treated with 0.05, 0.1, 0.2, 0.4, 0.8, 2.0 and 5.0 μmol / L final concentrations of 0 (solvent control, 0.1% DMSO) (DFO, 1 mmol / L) and MC-LR (0.8 μmol / L) and DFO (1 mmol / L) for 24 h. MTT ) Method to detect cell growth. The density of HT17 cells was adjusted to 1 × 10 5 / well, and the cells were treated with 0.1 mmol / L DFO (1 mmol / L) and MCAO (0.8 mmol / L) /L)+MC-LR(0.8μmol/L) were cultured for 24,48 h. The cell cycle and apoptosis were detected by flow cytometry. Results The cell viability of MC-LR treated with high dose (≥0.8μmol / L) for 24 h was significantly lower than that of solvent control (P <0.01), and the survival rate of DFO + MC-LR treated group was higher than that of 0.8μmol / L MC-LR group (P <0.05). Compared with the solvent control group, the proportion of cells in the G0 / G1 phase and the apoptosis rate increased after MC-LR exposure for 24 and 48 h, and the proportion of cells in S phase decreased (P <0.05 or P <0.01). There was no difference in the proportion of cells between DFO + MC-LR and MC-LR groups (P> 0.05). The apoptosis rate of DFO + MC-LR treated group was lower than that of MC-LR treated group (P <0.01) 24 h after exposure. Conclusion MC-LR can cause HT-17 cells to arrest and apoptosis in G0 / G1 phase, which may be related to the disorder of oxidative stress and protein phosphorylation induced by MC-LR.