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首页> 美国卫生研究院文献>World Journal of Gastroenterology >Lithium inhibits proliferation of human esophageal cancer cell line Eca-109 by inducing a G2/M cell cycle arrest
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Lithium inhibits proliferation of human esophageal cancer cell line Eca-109 by inducing a G2/M cell cycle arrest

机译：锂通过诱导G2 / M细胞周期阻滞抑制人食道癌细胞Eca-109的增殖

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AIM: To investigate the effect of lithium on proliferation of esophageal cancer (EC) cells and its preliminary mechanisms.METHODS: Eca-109 cells were treated with lithium chloride, a highly selective inhibitor of glycogen synthase kinase 3β (GSK-3β), at different concen-trations (2-30 mmol/L) and time points (0, 2, 4, 6 and 24 h). Cell proliferative ability was evaluated by 3-(4,5-dimethylthiazole-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and cell cycle distribution was examined by flow cytometry. Expressions of p-GSK-3β, β-catenin, cyclin B1, cdc2 and cyclin D1 protein were detected by Western blotting, and the subcellular localization of β-catenin was determined by immunofluorescence. The mRNA level of cyclin B1 was detected by reverse transcription polymerase chain reaction (RT-PCR).RESULTS: Lithium could inhibit the proliferation of Eca-109 cells. Lithium at a concentration of 20 mmol/L lithium for 24 h produced obvious changes in the distribution of cell cycle, and increased the number of cells in G2/M phase (P < 0.05 vs control group). Western blotting showed that lithium inhibited GSK-3β by Ser-9 phosphorylation and stabilized free β-catenin in the cytoplasm. Immunofluorescence further confirmed that free β-catenin actively translocated to the nucleus. Moreover, lithium slightly elevated cyclin D1 protein expression, whereas lowered the cyclin B1 expression after 24 h lithium exposure and no obvious change was observed for cdc2 protein.CONCLUSION: Lithium can inhibit the proliferation of human esophageal cancer cell line Eca-109 by inducing a G2/M cell cycle arrest, which is mainly mediated through the inhibition of lithium-sensitive molecule, GSK-3β, and reduction of cyclin B1 expression.
机译：目的：研究锂对食管癌细胞增殖的影响及其初步机制。方法：在Eca-109细胞中，用一种选择性高的糖原合酶激酶3β（GSK-3β）抑制剂氯化锂处理。不同浓度（2-30 mmol / L）和时间点（0、2、4、6和24小时）。通过3-（4，5-二甲基噻唑-2-基）-2、5-二苯基溴化四溴化铵（MTT）测定法评价细胞增殖能力，并通过流式细胞术检查细胞周期分布。 Western blot检测p-GSK-3β，β-catenin，cyclin B1，cdc2和cyclin D1蛋白的表达，并通过免疫荧光法检测β-catenin的亚细胞定位。通过逆转录聚合酶链反应（RT-PCR）检测细胞周期蛋白B1的mRNA水平。结果：锂可以抑制Eca-109细胞的增殖。锂浓度为20 mmol / L的锂24 h产生了明显的细胞周期分布变化，并增加了G2 / M期的细胞数量（与对照组相比，P <0.05）。 Western印迹表明，锂通过Ser-9磷酸化抑制了GSK-3β，并稳定了细胞质中的游离β-连环蛋白。免疫荧光进一步证实游离的β-连环蛋白活跃地转移到细胞核。此外，锂暴露24 h后，细胞周期蛋白D1蛋白表达略有升高，而细胞周期蛋白B1蛋白表达降低，而cdc2蛋白未见明显变化。结论：锂可通过诱导人食管癌细胞Eca-109增殖而抑制其增殖。 G2 / M细胞周期停滞，主要是通过抑制锂敏感分子GSK-3β和减少细胞周期蛋白B1表达来介导的。

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