实用医学杂志 ›› 2022, Vol. 38 ›› Issue (24): 3054-3058.doi: 10.3969/j.issn.1006⁃5725.2022.24.006

• 基础研究 • 上一篇    下一篇

METTL3介导IGF2BP3 m6A修饰对儿童肝母细胞瘤细胞增殖、迁移的影响 

魏慧珍1 允文晶2 崔西春3    

  1. 郑州大学第一附属医院 1 儿童重症监护室，2 综合重症监护室，3 小儿外科（郑州 450000）

  • 出版日期:2022-12-25 发布日期:2022-12-25
  • 基金资助:
    河南省卫生健康委员会基金项目（编号：LH⁃GJ20200354）

Effects of METTL3 mediated IGF2BP3 m6A modification on proliferation and migration of hepatoblasto⁃ ma cells in children

WEI Huizhen*，YUN Wenjing，CUI Xichun.   

  1. Children′s Intensive Care Unit of the First Affili⁃ ated Hospital of Zhengzhou University，Zhengzhou 450000，China

  • Online:2022-12-25 Published:2022-12-25

摘要：

目的 分析 METTL3 介导 IGF2BP3 m6A 修饰对儿童肝母细胞瘤细胞增殖、迁移的影响，为临床治疗应用提供依据。方法 采用 qRT⁃PCR 检测小儿肝母细胞瘤 HuH⁃6 中 METTL3 和 IGF2BP3 mRNA 的 表达；Western blot 检测 METTL3 和 IGF2BP3 蛋白表达。通过慢病毒转染，在 HuH⁃6 细胞中敲低 METTL3，并 通过免疫印迹验证METTL3蛋白表达，采用qRT⁃PCR 与Western blot 检测IGF2BP3的表达，m6A 甲基化RNA 免疫沉淀技术分析 IGF2BP3 mRNA m6A 修饰的情况；CCK⁃8 和 Transwell 小室实验检测细胞增殖和迁移能 力。结果 采用 METTL3 sgRNA 干预后会显著降低细胞 METTL3 表达水平（P < 0.05）；sgRNA 组细胞 48 h 和 72 h 时细胞增殖水平明显低于空载体组（P < 0.05）；sgRNA 组细胞 24 h 和 48 h 时细胞迁移明显低于空 载体组，sgRNA 组细胞侵袭水平明显低于空载体组；sgRNA 组细胞中 IGF2BP3 蛋白及 mRNA 表达水平明显 低于空载体组，且差异均有统计学意义（P < 0.05）；sgRNA 组细胞中 m6A⁃IP 及 IGF2BP3 m6A 相对表达量明 显低于空载体组，且差异均有统计学意义（P < 0.05）。结论 METTL3可有效调节IGF2BP3 m6A 修饰并对儿 童肝母细胞瘤细胞增殖、迁移进行调控。

关键词:

METTL3, IGF2BP3 m6A 修饰, 肝母细胞瘤, 细胞增殖, 细胞迁移, 儿童

Abstract:

Objective To analyze the effect of METTL3 mediated IGF2BP3 m6A modification on the proliferation and migration of hepatoblastoma cells in children，and provide basis for clinical treatment. Methods the expression of METTL3 and IGF2BP3 mRNA in children hepatoblastoma HuH ⁃6 was detected by QRT PCR ； Western blot was used to detect the expression of METTL3 and IGF2BP3. By lentiviral transfection，METTL3 was knocked down in HuH⁃6 cells，and the expression of METTL3 protein was verified by immunoblotting. The expres⁃ sion of IGF2BP3 was detected by QRT PCR and Western blot，and the m6A methylation RNA immunoprecipitation technique was used to analyze the m6A modification of IGF2BP3 mRNA；The cell proliferation ability was detected by CCK⁃8 assay，and the cell migration ability was detected by Transwell cell assay. Results METTL3 sgRNA intervention significantly decreased the expression level of METTL3 in cells，and the difference was statistically significant（P < 0.05）；The cell proliferation level of sgRNA group was significantly lower than that of empty vector group at 48 h and 72 h，and the difference was statistically significant（P < 0.05）；The cell migration of sgRNA group was significantly lower than that of empty vector group at 24 h and 48 h，and the cell invasion level of sgRNA group was significantly lower than that of empty vector group；The expression levels of IGF2BP3 protein and mRNA in sgRNA group were significantly lower than that in empty vector group（P < 0.05）；The relative expression of m6A IP and IGF2BP3 m6A in sgRNA group was significantly lower than that in empty vector group （P < 0.05）. Conclusion METTL3 can effectively regulate the modification of IGF2BP3 and m6A and regulate the proliferation and migration of hepatoblastoma cells in children.

Key words:

METTL3, IGF2BP3 m6A modification, hepatoblastoma, cell proliferation, cell migration, children

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