MiR-155对HuR的调控与结肠癌细胞的转移机制研究

doi:10.3969/j.issn.1006-6233.2023.11.06

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摘要 目的: 探讨MiR-155通过对HuR的调控参与结肠癌细胞转移的机制。方法: 研究使用HCT-116结肠癌细胞系,分为miR-155模拟物和miR-155抑制剂、miRNA模拟物阴性对照和miRNA抑制物阴性对照组。MTT、伤口愈合试验、菌落形成测定、细胞周期分析和流式细胞术测定检测细胞活力、周期、迁移和凋亡。qPCR和蛋白质印迹法测量细胞中细胞miR-155的表达水平,蛋白质印迹法测量细胞中细胞HuR的表达水平,荧光素酶报告基因测定相关靶点。结果: 与对照组比较,用miR-155模拟物转染后,miR-155水平升高;转染miR-155模拟物72h后细胞活力增加,菌落数增加,miR-155模拟物增加了S期细胞的百分比,miR-155模拟物抑制结肠癌细胞的凋亡,转染miR-155模拟物的细胞的迁移率增加,明确miR-155模拟物促进了结肠癌细胞的迁移。用miR-155抑制剂转染后,结果相反。用miR-155抑制剂转染后HuR的mRNA水平降低,转染miR-155模拟物后,HuR的mRNA水平升高。荧光素酶报告基因测定miR-155与HuR的3′UTR结合,并且HuR是miR-155的直接靶标。结论: 结果表明miR-155促进结肠癌细胞增殖,增强其集落形成能力,促进其细胞周期进程,抑制细胞凋亡,促进了结肠癌细胞的迁移。HuR被确定为miR-155的新靶点。

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关键词 ： miR-155, HuR, 结肠癌, 转移

Abstract：Objective: To explore the mechanism of miR-155 involved in colon cancer cell metastasis through the regulation of HuR. Methods: HCT-116 colon cancer cell lines were used in the research.They were categorized into groups with miR-155 mimics,miR-155 inhibitors,miRNA mimics as a negative control,and miRNA inhibitors as a negative control.Cell viability,cell cycle,migration,and apoptosis were assessed using MTT,wound healing tests,colony formation assays,cell cycle analysis,and flow cytometry.The expression of miR-155 in cells was measured using qPCR and western blotting.The expression of HuR in cells was also measured using western blotting.Relevant targets were determined using a luciferase reporter gene. Results: Compared to the control group,the level of miR-155 increased after transfection with miR-155 mimics.After 72 hours of transfection with miR-155 mimics,cell activity and colony formation increased.The percentage of S-phase cells increased,and apoptosis was inhibited by miR-155 mimics in colon cancer cells.MiR-155 mimics also increased cell mobility,indicating they promoted the migration of colon cancer cells.These results were reversed after transfection with miR-155 inhibitors.The mRNA level of HuR decreased after transfection with miR-155 inhibitors and increased after transfection with miR-155 mimics.The luciferase reporter gene confirmed the binding of miR-155 to the 3'UTR of HuR,establishing HuR as a direct target of miR-155. Conclusion: The findings suggest that miR-155 promotes colon cancer cell proliferation,enhances colony formation,drives the cell cycle process,inhibits apoptosis,and increases cell migration.HuR is identified as a new target of miR-155.

Key words： miR-155 HuR Colon cancer Metastasis

基金资助:湖北省荆门市科学技术研究与开发计划,(编号:2022YFYB042);湖北省卫生健康科研基金项目,(编号:H20220036)

通讯作者: 强勇

引用本文:

张小风, 赵建锋, 吕洋, 范金强, 强勇. MiR-155对HuR的调控与结肠癌细胞的转移机制研究[J]. 河北医学, 2023, 29(11): 1789-1794.
ZHANG Xiaofeng, ZHAO Jianfeng, LV Yang, et al. The Metastasis Mechanism of Colon Cancer Cells Through the Regulation of HuR by MiR-155. HeBei Med, 2023, 29(11): 1789-1794.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2023.11.06 或 http://www.hbyxzzs.cn/CN/Y2023/V29/I11/1789

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