实用医学杂志 ›› 2023, Vol. 39 ›› Issue (13): 1627-1633.doi: 10.3969/j.issn.1006⁃5725.2023.13.006

• 基础研究 • 上一篇    下一篇

miR⁃30b⁃5p通过下调MKRN3表达抑制食管癌细胞增殖、迁移和侵袭 

周乾华 毕华俊 孟凡东 施益明 芮超    

  1. 安徽省第二人民医院胸心外科(合肥 230012) 
  • 出版日期:2023-07-10 发布日期:2023-07-10
  • 基金资助:
    安徽省卫生健康委科研计划项目(编号:2018SEYL024)

MiR⁃30b⁃5p inhibits the proliferation,migration and invasion of esophageal cancer cells by down⁃regulat⁃ ing the expression of MKRN3 

ZHOU Qianhua,BI Huajun,MENG Fandong,SHI Yiming,RUI Chao.    

  1. Depart⁃ ment of Thoracic and Cardiovascular Surgery,Anhui Second People′s Hospital,Hefei 233000,China 
  • Online:2023-07-10 Published:2023-07-10

摘要:

目的 探讨 miR⁃30b⁃5p 在食管癌细胞中的表达及对食管癌细胞增殖、迁移和侵袭的影响机制。方法 收集 2016 年 1 月至 2020 年 12 月就诊于安徽省第二人民医院手术治疗的 70 例食管癌患者的肿 瘤和癌旁组织,检测 miR⁃30b⁃5p 的表达水平。qRT⁃PCR 检测食管癌细胞株中 miR⁃30b⁃5p 的表达水平。将 miR⁃30b⁃5p mimics转染至TE⁃13和ECA109细胞中,检测转染后各组细胞中miR⁃30b⁃5p及MKRN3表达水平。 根据细胞增殖实验,Transwell 及裸鼠成瘤实验检测食管癌细胞增殖、迁移及侵袭能力。双荧光素酶实验验 证 miR⁃30b⁃5p 的靶基因。蛋白印迹实验探索 miR⁃30b⁃5p 对食管癌影响的分子机制。结果 miR⁃30b⁃5p 在食管癌细胞和食管癌组织中相较于正常上皮细胞和癌旁组织表达显著下降(P < 0.05)。细胞增殖实验显示, miR⁃30b⁃5p mimics 组增殖率显著低于 NC 组(P < 0.05);Transwell 实验结果显示,miR⁃30b⁃5p mimics 组迁移 率及侵袭率显著低于NC组(P < 0.05);双荧光素酶实验显示MKRN3是miR⁃30b⁃5p的直接靶基因。Western blot检测JAK/STAT 信号通路蛋白发现,miR⁃30b⁃5p mimics组蛋白低于NC组(P < 0.05)。结论 miR⁃30b⁃5p 在食管癌中呈现低表达,miR⁃30b⁃5p通过下调MKRN3表达抑制食管癌细胞增殖、迁移和侵袭能力。 

关键词: miR?30b?5p, makorin 环指蛋白 3, 食管癌, JAK/STAT 信号通路, 细胞增殖, 细胞 迁移, 细胞侵袭

Abstract:

Objective To investigate the expression of miR⁃30b⁃5p in esophageal cancer cells and its effect on the proliferation,migration,and invasion of esophageal cancer cells. Methods The expression level of miR⁃30b⁃5p was detected in tumor and adjacent tissues of 70 patients with esophageal cancer who received surgical treatment in our hospital from January 2016 to December 2020. The expression level of miR⁃30b⁃5p in esophageal cancer cell lines was detected by qRT⁃PCR. miR⁃30b⁃5p mimics were transfected into TE⁃13 and ECA109 cells, and the expression levels of miR⁃30b⁃5p and MKRN3 in the transfected cells were detected. The proliferation, migration and invasion ability of esophageal carcinoma cells were determined by cell proliferation assay,Transwell assay and nude mouse tumorigenesis assay. The target gene of miR⁃30b⁃5p was verified by double luciferase assay. Western blotting was used to explore the molecular mechanism of the effect of miR⁃30b⁃5p on esophageal carcino⁃ ma. Results The expression of miR⁃30b⁃5p in esophageal carcinoma cells and tissues was significantly decreased compared with normal epithelial cells and paracancer tissues(P < 0.05). Cell proliferation experiment showed that the proliferation rate of miR⁃30b⁃5p mimics group was significantly lower than that of NC group(P < 0.05). Tran⁃ swell experiment results showed that the mobility and invasion rates of miR⁃30b⁃5p mimics group were significantly lower than those of NC group(P < 0.05). Dual luciferase assay showed that MKRN3 was the direct target gene of miR⁃30b⁃5p. Western blot analysis of JAK/STAT signaling pathway protein showed that the histone of miR⁃30b⁃5p mimics was lower than that of NC group(P < 0.05). Conclusion miR⁃30b⁃5p shows low expression in esophageal cancer,and miR⁃30b⁃5p inhibits the proliferation,migration and invasion of esophageal cancer cells by down⁃regu⁃ lating the expression of MKRN3.

Key words: miR ?30b ?5p; makorin ring finger protein 3; esophageal cancer; JAK/STAT signaling pathway; cell migration; cell invasion ,