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中国癌症防治杂志 ›› 2015, Vol. 7 ›› Issue (1): 12-17.doi: 10.3969/j.issn.1674-5671.2015.01.03

• 基础研究 • 上一篇    下一篇

二甲双胍对LKB1基因过表达的子宫内膜癌HEC-1A细胞的影响

  

  1. 广西医科大学附属肿瘤医院妇瘤二科;广西医科大学研究生学院 
  • 出版日期:2015-02-25 发布日期:2015-02-28
  • 通讯作者: 宋红林。E-mail:381783170@qq.com
  • 基金资助:

    广西自然科学基金资助项目(2011GXNSFA018253)

Effect of metformin on HEC-1A endometrial cancer cells in the presence of LKB1 overex-pression

  • Online:2015-02-25 Published:2015-02-28

摘要:

目的 研究二甲双胍对LKB1基因过表达的子宫内膜癌HEC-1A细胞的增殖与细胞周期的影响。方法 以不同浓度的二甲双胍作用于LKB1基因过表达的子宫内膜癌HEC-1A细胞(LKB组)与缺失LKB1基因表达的HEC-1A细胞(CON组)。用流式细胞术仪检测各组HEC-1A细胞的细胞周期;RT-PCR法和Western blot技术检测各组HEC-1A细胞的LKB1、mTOR基因与蛋白的表达;以平板克隆法与细胞迁移实验检测各组细胞的细胞增殖水平。结果 与缺失LKB1基因表达的HEC-1A细胞比较,LKB1基因过表达的HEC-1A细胞随着二甲双胍浓度的升高明显降低细胞克隆形成率、细胞迁移率、S 期细胞比例及mTOR基因与蛋白的表达,而增加G0 /G1期细胞比例及LKB1基因与蛋白的表达,差异均有统计学意义(P均<0.05)。结论 二甲双胍抑制LKB1基因过表达的子宫内膜癌HEC-1A细胞的生长和侵袭,其机制可能为介导LKB1/mTOR信号传导通路而抑制子宫内膜癌细胞的增殖。

关键词: 子宫肿瘤, 肝激酶基因, HEC-1A细胞, 二甲双胍, 胰岛素抵抗

Abstract:

Objective To investigate the effect of metformin on proliferation and cell cycle of HEC-1A endometrial cancer cells in the presence of LKB1 overexpression. Methods HEC-1A cells overexpressing LKB1 and control HEC-1A cells were treated with different concentrations of metformin. The two groups were compared in terms of proliferation rate,cell migration ability and cell cycle using a colony formation assay,transwell experiments and flow cytometry. Expression of LKB1 and mTOR mRNA was measured using RT-PCR,and levels of the corresponding proteins were measured using Western blotting. Results Treating LKB1-overexpressing HEC-1A cells with metformin inhibited colony formation,cell motility,and expression of mTOR mRNA and protein in a dose-dependent manner relative to control HEC-1A cells(P<0.05). It also reduced the proportion of LKB1-overexpressing HEC-1A cells in S phase relative to controls(P<0.05). Conversely,treating LKB1-overexpressing HEC-1A cells with metformin increased the proportion of cells in G0 /G1 phase and upregulated expression of LKB1 mRNA and protein in a dose-dependent manner(P<0.05). Conclusion Metformin inhibits proliferation and cell migration of HEC-1A endometrial cancer cells overexpressing LKB1,potentially by acting on the LKB1/mTOR signaling pathway.

Key words: Endometria, neoplasm, Liver kinase B1, HEC-1A cell, Metformin, Insulin resistance