文章摘要
丁敬健,张升涛,郭永锋,等.微 RNA-196a-1-3p靶向 Ras响应元件结合蛋白调控胆管癌细胞增殖的机制研究[J].安徽医药,2024,28(7):1399-1403.
微 RNA-196a-1-3p靶向 Ras响应元件结合蛋白调控胆管癌细胞增殖的机制研究
Mechanism of miR-196a-1-3p targeting RREB1 to regulate the proliferation of cholangiocarcinoma cells
  
DOI:10.3969/j.issn.1009-6469.2024.07.028
中文关键词: 胆管肿瘤  转化生长因子 β  细胞增殖  微 RNA-196a-1-3p  Ras反应元件结合蛋白 1  SMAD家族成员 3
英文关键词: Bile duct neoplasms  Transforming growth factor beta  Cell proliferation  MiR-196a-1-3p  Ras responsive element binding protein 1  SMAD family member 3
基金项目:西安市卫生厅科研基金项目( 20A038)
作者单位E-mail
丁敬健 西安市第九医院普通外科陕西西安710054  
张升涛 西安市第九医院普通外科陕西西安710054  
郭永锋 西安市第九医院普通外科陕西西安710054  
王尚毓 空军军医大学第一附属医院肝胆外科陕西西安 710032  
罗孔亮 西安市第九医院普通外科陕西西安710054  
董伟 西电集团医院普通外科陕西西安 710077 304145789@qq.com 
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中文摘要:
      目的探讨转化生长因子 β(TGF-β)调控人胆管癌细胞系 RBE细胞增殖的关键微 RNA(miRNA)及其潜在的机制。方法该研究起止时间为 2020年 1月至 2022年 1月。磷酸盐缓冲液( PBS)处理为对照组, TGF-β处理为 TGF-β组, TGF-β抗体处理为抗体组。检测三组 RBE细胞的增殖水平。 miRNA高通量测序检测三组 RBE细胞的 miRNA调控变化,并进行 miRNA模拟物过表达筛选鉴定受 TGF-β调控的影响 RBE细胞增殖水平的关键 miRNA。miRNA数据库( miRDB)在线分析 miRNA的潜在底物,并通过小干扰 RNA(siRNA)敲低筛选鉴定影响 RBE细胞增殖水平的关键底物。结果相比于对照组, TGF-β组 RBE细胞的增殖水平上升( 1.62±0.07比 2.35±0.09,P<0.05)抗体组 RBE细胞的增殖水平下降( 1.62±0.07比 1.11±0.08,P<0.05)。过表达微 RNA-196a-1-3p(miR-196a-1-3p)时, RBE细胞的增,殖水平下降( P<0.05)。敲低 Ras响应元件结合蛋白( RREB1)时, RBE细胞的增殖水平下降( P<0.05)。过表达 miR-196a-1-3p后, RBE细胞中 RREB1的信使 RNA(mRNA)和蛋白水平下降( P<0.05)。敲低 miR-196a-1-3p后, RBE细胞中 RREB1与 SMAD家族蛋白 3(SMAD3)的相互作用增加。敲低 SMAD3后, RBE细胞的增殖水平下降( P<0.05)。与仅敲低 SMAD3相比,敲低 SMAD3的同时过表达 RREB1的 RBE细胞的增殖水平无显著变化,并且同时敲低 SMAD3和 miR-196a-1-3p的 RBE细胞的增殖水平无显著变化。结论 TGF-β能够通过 miR-196a-1-3p/RREB1/SMAD3轴促进 RBE细胞增殖; miR-196a-1-3p和 RREB1可作为潜在的治疗胆管癌的靶标,为针对该靶标的新药研发奠定了基础。
英文摘要:
      Objective To investigate the key microRNAs (miRNAs) of transforming growth factor β (TGF-β) regulating cholangiocarcinoma cells (RBE) proliferation and their potential mechanisms.Methods The starting and ending time of this study was from January 2020 to January 2022. Phosphate buffer (PBS) treatment was used as the control group, TGF-β treatment as the TGF-β group, and TGF-β antibody treatment as the antibody group. The proliferation levels of RBE cells in the three groups were detected. miRNA high-throughput sequencing (miRNA-seq) was performed to detect miRNA regulatory changes in the three groups of RBE cells, and miRNAmimics overexpression screening was performed to identify key miRNAs regulated by TGF-β that affect the proliferation levels of RBEcells. miRNA database (miRDB) was analyzed online for potential substrates of miRNAs, and small interfering RNA (siRNA) knockdown screen to identify key substrates affecting the proliferation level of RBE cells.Results Compared to the control group, the proliferation level of RBE cells was increased in the TGF-β group (1.62±0.07 vs. 2.35±0.09, P<0.05) and decreased in the antibody group (1.62±0.07 vs. 1.11±0.08, P<0.05). The proliferation level of RBE cells was decreased upon overexpression of miR-196a-1-3p (P< 0.05). The proliferation level of RBE cells was decreased when Ras response element binding protein (RREB1) was knocked down (P< 0.05). The messenger RNA (mRNA) and protein levels of RREB1 were decreased in RBE cells after overexpression of miR-196a-1-3p (P<0.05). The interaction of RREB1 with SMAD3 was increased in RBE cells after knockdown of miR-196a-1-3p. The proliferation lev·el of RBE cells decreased after knockdown of SMAD3 (P<0.05). There was no significant change in the proliferation level of RBE cellswith knockdown of SMAD3 and overexpression of RREB1 compared to knockdown of SMAD3 alone, and there was no significantchange in the proliferation level of RBE cells with both SMAD3 and miR-196a-1-3p knockdown. Conclusion TGF-β can promote RBE cell proliferation through miR-196a-1-3p/RREB1/SMAD3 axis. miR-196a-1-3p and RREB1 can be used as potential targets forthe treatment of cholangiocarcinoma, laying the foundation for the development of new drugs against this target.
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