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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (4): 340-345.doi: 10.12047/j.cjap.5935.2020.073

• 研究论文 • 上一篇    下一篇

miR-520a-3p调控宫颈癌细胞因子分泌的信号通路*

刘红艳1, 鲁启洪2△, 黄楚鹰3   

  1. 1.湖北省恩施土家族苗族自治州中心医院 肿瘤放疗中心, 恩施 445000;
    2.湖北省恩施土家族苗族自治州中心医院神经内科, 恩施 445000;
    3.湖北省恩施土家族苗族自治州中心医院肿瘤一科, 恩施 445000
  • 收稿日期:2019-09-09 修回日期:2020-05-19 发布日期:2020-11-09
  • 通讯作者: Tel: 13997794720; E-mail: wxl15608070633@163.com

miR-520a-3p regulates the signaling pathway of cytokine secretion in cervical cancer

LIU Hong-yan1, LU Qi-hong2△, HUANG Chu-ying3   

  1. 1. Tumor Radiotherapy Center of Central Hospital of Enshi Tujia Miao Autonomous Prefecture, Hubei Province, Enshi 445000;
    2. CentralHospital of Enshi Tujia and Miao Autonomous Prefecture, Hubei Province, Department of Neurology, Enshi 445000;
    3. Department of Oncology, Central Hospital of Enshi Tujia and Miao Autonomous Prefectures, Hubei Province, Enshi 445000, China
  • Received:2019-09-09 Revised:2020-05-19 Published:2020-11-09

摘要: 目的: 探讨miR-520a-3p调控宫颈癌细胞因子分泌的分子机制。方法: 通过TargetScanHuman分析miR-520a-3p与NF-κB复合体亚基RELA的匹配情况,然后通过荧光素酶报告系统检测miR-520a-3p是否靶向NF-κB复合体亚基RELA;使用LPS刺激宫颈癌HELA细胞后,将miR-520a-3p mimics与转染试剂混合后滴入HELA细胞中,此为过表达组;将miR-520a-3p inhibitor与转染试剂混合后滴入HELA细胞中,此为敲低组,通过酶联免疫吸附试验检测过表达组和敲低组GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα的表达水平。每次实验重复3次。结果: miR-520a-3p靶向RELA的3’UTR;LPS激活NF-kB信号通路后,宫颈癌HELA细胞分泌的细胞因子GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα的蛋白表达水平上升(P<0.05);过表达组中NF-κB复合体亚基RELA的蛋白表达水平下降,宫颈癌HELA细胞分泌的上述细胞因子的蛋白表达水平下降(P<0.05);敲低组中NF-κB复合体亚基RELA的蛋白表达水平上升,宫颈癌HELA细胞分泌的上述细胞因子的蛋白表达水平上升(P<0.05)。结论: miR-520a-3p通过靶向NF-κB信号通路的关键分子RELA抑制宫颈癌HELA细胞的细胞因子分泌。

关键词: miR-520a-3p, NF-kB, 宫颈癌HELA细胞, 细胞因子

Abstract: Objective: To investigate the molecular mechanism of miR-520a-3p regulating the secretion of cytokines in cervical cancer. Methods: The matching between miR-520a-3p and the NF-κB complex subunit RELA was analyzed by TargetScanHuman, and then the luciferase reporting system was used to detect whether miR-520a-3p targeted the NF-κB complex subunit RELA. After cervical cancer HELA cells stimulated by LPS, in the overexpression group or the knockout group, miR-520a-3p mimics or inhibitor were mixed with transfection reagent and dripped into HELA cells. The expression levels of colony stimulating factor (GCSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 2 (IL-2), interleukin 3 (IL-3), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 19 (IL-9), interleukin 10 (IL-10), interleukin 12 p40 (IL-12 p40), interleukin 12 p70 (IL-12 p70), interleukin 13 (IL-13), interleukin 17 (IL-17), interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), monocyte chemoattractant protein-5 (MCP-5), small inducible cytokine A5 (RANTES), tumor necrosis factor-alpha (TNFα) were detected by enzyme-linked immunosorbent assay in both the overexpression group and the knockout group. Each experiment was repeated three times. Results: miR-520a-3p targeted the 3’UTR of RELA. After activation of the NF-κB signaling pathway by lipopolysaccharide (LPS), the protein expression levels of cytokines GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα secreted by HELA cells of cervical cancer were increased significantly (P<0.05). In the overexpression group, the protein expression level of RELA of NF-κB complex subunit was decreased, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were decreased (P<0.05). In the knockdown group, the protein expression level of RELA, a subunit of NF-κB complex, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were increased (P<0.05). Conclusion: miR-520a-3p inhibits cytokine secretion in HELA cells of cervical cancer by targeting RELA, a key molecule in the NF-κB signaling pathway.

Key words: miR-520a-3p, NF-κB, HELA cells of cervical cancer, cytokines

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