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中国癌症防治杂志

中国癌症防治杂志 ›› 2021, Vol. 13 ›› Issue (1): 45-50.doi: 10.3969/j.issn.1674-5671.2021.01.08

• 基础研究 • 上一篇    下一篇

miR-106b对肝癌细胞放疗敏感性的影响及其作用机制

  

  1. 淮安市第二人民医院肝胆外科
  • 出版日期:2021-02-25 发布日期:2021-03-05
  • 通讯作者: 王正 E-mailwz110518@163.com
  • 基金资助:
    2018年高层次卫生人才“六个一工程”拔尖人才科研项目（LGY2018043）

Effect of miR-106b on radiosensitivity of hepatoma cells and its mechanism

  • Online:2021-02-25 Published:2021-03-05

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摘要： 目的 探讨miR-106b对肝癌细胞放疗敏感性的影响及其可能的作用机制。方法 采用qRT-PCR检测人肝癌细胞株HepG2、SMMC-7721、SK-HEP-1、Huh7及正常肝细胞株QSG7701中miR-106b mRNA的表达。用miR-106b siRNA转染HepG2细胞（miR-106b下调组），并设空白对照组和阴性对照组。不同剂量（0 Gy、2 Gy、4 Gy、6 Gy、8 Gy）照射后，采用细胞克隆形成实验检测各组细胞克隆形成率（plating efficiency，PE）及细胞存活分数（survival fraction，SF）；6 Gy照射后，采用CCK-8法、流式细胞仪分别检测各组细胞增殖、凋亡情况，Western blot检测p-PI3K/PI3K、p-AKT/AKT、PCNA和Bax蛋白表达情况。结果 与QSG7701细胞相比，HepG2、SMMC-7721、SK-HEP-1、Huh7细胞中miR-106b表达水平均升高（均P＜0.05），其中HepG2细胞中miR-106b的表达水平最高。克隆形成实验结果显示，不同剂量（0 Gy、2 Gy、4 Gy、6 Gy、8 Gy）照射后，HepG2细胞的PE、SF水平随照射剂量升高而逐渐降低（均P＜0.05），其中6 Gy和8 Gy照射的PE、SF水平差异无统计学意义（均P＞0.05）。6 Gy照射后，与空白对照组、阴性对照组相比，miR-106b下调组HepG2细胞的增殖能力及p-PI3K/PI3K、p-AKT/AKT、PCNA蛋白表达水平均降低（均P＜0.05），细胞凋亡率及Bax蛋白表达水平则升高（均P＜0.05）。结论 下调miR-106b表达可抑制HepG2细胞增殖并诱导凋亡，增强放疗敏感性，其作用机制可能与阻断PI3K/AKT通路活化有关。

关键词: 肝癌, miR-106b, 放疗敏感性, PI3K/AKT通路

Abstract: Objective To investigate the effect of miR-106b on the radiosensitivity of hepatoma cells and its possible mechanism. Methods The expression of miR-106b in human hepatoma cell lines HepG2, SMMC-7721, SK-HEP-1, Huh7 and normal hepatocyte QSG7701 were detected by qRT-PCR. The HepG2 cells were transfected with miR-106b siRNA(miR-106b down-regulated group), and the blank control group and negative control group were set up. After irradiated by different doses(0 Gy, 2 Gy, 4 Gy, 6 Gy, 8 Gy), cell plating efficiency(PE) and cell survival fraction(SF) were determined by cell clone formation assay. After irradiated by 6 Gy, the proliferation and apoptosis of cells in each group were detected by CCK-8 method and flow cytometry; the protein expressions of p-PI3K/PI3K, p-AKT/AKT, PCNA and Bax were detected by Western blot. Results Compared with QSG7701 cells, the expression level of miR-106b in HepG2, SMMC-7721, SK-HEP-1, and Huh7 cells was increased(all P<0.05), and the expression level of miR-106b in HepG2 cells was the highest. The results of clone formation experiment showed that PE and SF levels in HepG2 cells were reduced with the increase of irradiation dose (all P<0.05) after irradiated by different doses(0 Gy, 2 Gy, 4 Gy, 6 Gy, 8 Gy), and there was no statistical significance in PE and SF levels between 6 Gy and 8 Gy irradiation (both P>0.05). After 6 Gy irradiation , compared with the blank control group and the negative control group, the proliferation ability of HepG2 cells and the protein expression levels of p-PI3K/PI3K, p-AKT/AKT and PCNA in miR-106b down-regulated group were decreased(all P<0.05), while the apoptosis rate and the protein expression level of Bax were increased(all P<0.05). Conclusion The down-regulation of the expression of miR-106b can inhibit the proliferation and induce the apoptosis of HepG2 cells, while enhance the radiosensitivity, and the underlying mechanism may be related to the blocking of PI3K/AKT pathway activation.

Key words: Hepatoma, miR-106b, Radiosensitivity, PI3K/AKT pathway

中图分类号: 

  • R735.7