实用医学杂志 ›› 2022, Vol. 38 ›› Issue (2): 160-167.doi: 10.3969/j.issn.1006⁃5725.2022.02.007

• 基础研究 • 上一篇    下一篇

miR⁃26b⁃5p通过靶向NFE2L3调控结直肠癌细胞生长迁移和侵袭

黄波1 李慧雯2 常诚1   

  1. 1 广州市红十字会医院普外科（广州 510220）；2 广州市妇女儿童医疗中心消化内科（广州 510623）

  • 出版日期:2022-01-25 发布日期:2022-01-25

miR⁃26b⁃5p regulates the growth，migration and invasion of colorectal cancer cells by targeting NFE2L3

HUANG Bo*，LI Huiwen，CHANG Cheng   

  1. Department of General Surgery，Guangzhou Red Cross Hospital，Guang⁃ zhou 510220，China

  • Online:2022-01-25 Published:2022-01-25

摘要：

目的 探究 miRNA⁃26b⁃5p（miR⁃26b⁃5p）靶向调控 NFE2L3 表达水平影响结直肠癌细胞生长、 迁移和侵袭能力的分子机制。方法 通过 qPCR 的方法检测了人正常肠上皮细胞（NCM460）及结直肠癌 细胞系（HT29、SW620及HCT116）中miR⁃26b⁃5p的表达水平。通过qPCR及western blot检测了上述细胞系中 NFE2L3的表达水平。生物信息学方法分析了miRNA⁃26b⁃5p与NFE2L3结合的靶向序列。双荧光素酶基因报告实验检测 miR⁃26b⁃5p 对 NFE2L3 的靶向调控机制。用脂质体法将 miR⁃26b⁃5p mimic 或同时将 NFE2L3 转染 HT29及SW620结直肠癌细胞系中，通过CCK⁃8及克隆形成实验观察对细胞生长的影响，通过流式细胞术 观察凋亡能力的影响，通过Transwell观察细胞迁移和侵袭能力的影响。结果 与人正常上皮细胞系相比，在结直肠癌细胞系中miR⁃26b⁃5p的表达水平显著下调（P < 0.05），而NEF2L3的mRNA水平及蛋白水平则显著 上调（P < 0.05）。双荧光素酶报道基因实验显示miR⁃26b⁃5p能显著影响NEF2L3 3′UTR 表达载体的荧光素 酶活性（P < 0.05）。过表达miR⁃26b⁃5p可显著抑制结直肠癌细胞系细胞的生长、克隆形成及迁移和侵袭能力， 引起细胞凋亡（均P < 0.05），而同时过表达NEF2L3则可抵消miR⁃26b⁃5p的对细胞功能的作用（均P < 0.05）。 结论 在结直肠癌中miR⁃26b⁃5p低表达，而NEF2L3高表达；miR⁃26b⁃5p可靶向调控癌基因EF2L3的表达水平，从而调控结直肠癌细胞的生长、凋亡、迁移和侵袭，参与结直肠癌的发生发展。

关键词: miR?26b?5p, NEF2L3, 结直肠癌, 生长, 凋亡, 迁移, 侵袭

Abstract:

Objective To explore the molecular mechanism that miRNA ⁃26b ⁃5p（miR ⁃26b ⁃5p）targets the expression level of NFE2L3 and affects the growth，migration，and invasion of colorectal cancer cells. Methods The expression levels of miR⁃26b⁃5p in human normal intestinal epithelial cells（NCM460）and colorectal cancer cell lines（HT29，SW620 and HCT116）were detected by qPCR. The expression level of NFE2L3 in the above cell lines was detected by qPCR and western blot. Bioinformatics methods analyzed the target sequence of miRNA⁃26b⁃ 5p binding to NFE2L3. The dual luciferase gene reporter experiment detects the targeted regulation mechanism of miR⁃26b⁃5p on NFE2L3. Liposome method was used to transfect miR⁃26b⁃5p mimic or NFE2L3 into HT29 and SW620 colorectal cancer cell lines，to observe the effect on cell growth through CCK⁃8 and clone formation experiment，to observe cell apoptosis by flow cytometry and to observe cell migration and invasion through Transwell assay. Results Compared with human normal epithelial cell lines，the expression level of miR⁃26b⁃5p in colorectal cancer cell lines was significantly down⁃regulated（P < 0.05），while the mRNA and protein levels of NEF2L3 were significantly up⁃regulated（P < 0.05）. The dual luciferase gene reporter experiment showed that miR⁃26b⁃5p can significantly affect the luciferase activity of NEF2L3 3′UTR expression vector（P < 0.05）. Overexpression of miR⁃ 26b⁃5p significantly inhibited the growth，colony formation，migration and invasion of colorectal cancer cell lines， and induce cell apoptosis（all P < 0.05），while overexpression of NEF2L3 can counteract the effects of miR⁃26b⁃5p （all P < 0.05）. Conclusion In colorectal cancer ，miR⁃26b⁃5p is lowly expressed ，while NEF2L3 is highly expressed；miR ⁃26b ⁃5p can target and regulate the expression level of oncogene EF2L3，thereby regulating the growth，apoptosis，migration and invasion of colorectal cancer cells，thereby participating in the tumorigenesis and development of colorectal cancer.

Key words:

miR?26b?5p, NEF2L3, colorectal cancer, growth, apoptosis, migration, invasion

粤ICP备18046265号
版权所有 © 2019 《实用医学杂志》编辑部
地址：广州市越秀区惠福西路进步里2号之6　
邮编：510180　电话：020-81866302；81872080；81840509；81922330　
E-mail： syyxzz@syyxzz.com
本系统由北京玛格泰克科技发展有限公司设计开发