实用医学杂志 ›› 2021, Vol. 37 ›› Issue (17): 2187-2193.doi: 10.3969/j.issn.1006⁃5725.2021.17.004

• 基础研究 • 上一篇    下一篇

miR⁃128⁃3p靶向调控ZEB1表达对结肠癌细胞侵袭和迁移能力的影响

马静1 滕圣敏1 李苓2   

  1. 徐州医科大学附属滕州市中心人民医院1 病理科,2 肿瘤科(山东滕州277500)

  • 出版日期:2021-09-10 发布日期:2021-09-10
  • 通讯作者: 李苓 E⁃mail:lilingtz@163.com
  • 基金资助:

    山东省肿瘤防治联合基金项目(编号:ZR2020LZL019


miR ⁃ 128⁃3p affects the invasion and migration of colorectal cancer cells by targeting ZEB1

MA Jing* TENG ShengminLI Ling.   

  1. Department of Pathology,Affiliated Tengzhou Central People′s Hospital of Xuzhou Medi⁃ cal University,Tengzhou 277500,China

  • Online:2021-09-10 Published:2021-09-10
  • Contact: LI Ling E⁃mail:lilingtz@163.com

摘要:

目的 探讨 miR⁃128⁃3p 在结直肠癌中的表达及其对结直肠癌细胞侵袭和迁移能力的影响 和可能的作用机制。方法 选取 2015 6 月至 2017 5 月我院手术切除的 56 例结直肠癌组织及癌旁组织标本,并收集结直肠癌细胞系和人正常结直肠黏膜细胞,采用 qPCR 法检测 miR⁃128⁃3p 相对表达水平并分析其与结直肠癌临床病理特征的关系。分别将 miR⁃128⁃3p mimic、mimic NC、miR⁃128⁃3p inhibitor inhibitor NC 转染至 SW620 细胞,采用 transwell 实验检测其侵袭和迁移能力的改变。采用双荧光素酶报告 基因实验验证 miR⁃128⁃3p ZEB1 的靶向关系,将 ZEB1 质粒单独或联合 miR⁃128⁃3p mimic 转染至 SW620 细胞,采用 transwell 实验检测其侵袭和迁移能力改变,分别采用 qQCR WB 实验检测 SW620 细胞中 ZEB1 蛋白及 mRNA 的表达。结果 miR⁃128⁃3p 在结直肠癌组织和结直肠癌细胞中低表达(均 P < 0.05),其表达水平与肿瘤分化程度、淋巴结转移和远处转移有关(均 P < 0.05);瞬时转染 miR⁃128⁃3p mimic 能显著抑制 SW620 细胞的侵袭和迁移,瞬时转染 miR⁃128⁃3p inhibitor 能显著增强 SW620 细胞的侵袭和迁移。双荧光 素酶报告基因实验证实 miR⁃128⁃3p 直接靶向调节 ZEB1 的表达,过表达 ZEB1 能促进 SW620 细胞的侵袭和 迁移,同时转染 miR⁃128⁃3p 可下调 ZEB1 蛋白和 mRNA 表达,SW620 细胞的侵袭和迁移能力下降。结论 miR⁃128⁃3p 在结直肠癌中低表达并与肿瘤分化程度、淋巴结转移和远处转移相关,miR⁃128⁃3p 通过靶向调ZEB1抑制结直肠癌的侵袭和迁移。

关键词:

结直肠癌, miR?128?3p, 转录因子E 盒结合蛋白1, 侵袭, 迁移 ,

Abstract:

Objective To investigate the expression of miR⁃128⁃3p in colorectal cancer(CRC)and its effect on the invasion and migration of CRC cells and the underlying mechanism. Methods The canceroustissues and paired para⁃cancerous tissues of CRC patients(n = 56)who underwent surgical treatment in our hospital from⁃ June 2015 to May 2017 were collected. CRC cell lines and human normal colorectal mucosal cells were obtained. The expression of miR⁃128⁃3p in CRC tissues and cells were detected by qPCR and its relationship with the clinico⁃ pathologic feature of CRC patient was further analyzed. miR⁃128⁃3p mimic,mimic NC,miR⁃128⁃3p inhibitor and inhibitor NC were transfected into SW620 cells,respectively. Transwell assay was used to detected the invasion and migration of SW620 cells. Luciferase reporter assay was used to validate the targeted relationship between miR⁃128⁃3p and ZEB1. After transfection with ZEB1plasmid alone or in combination with miR⁃128⁃3p mimics,the invasion and migration ability of SW620 cells was detected by transwell assay,the expression of ZEB1 protein or mRNA in SW620 cells was detected by WB and qPCR,respectively. Results miR⁃128⁃3p was down⁃regulated in CRC cells and tissues(P < 0.05),and its expression level was related to the degree of tumor differentiation lymph node metastasis and distant metastasis(P < 0.05). Up⁃regulated expression of miR⁃128⁃3pcould significantly inhibit the invasion and migration of SW620 cells and down⁃regulated expression of miR⁃128⁃3p reversed the inhibi⁃ tory effect of miR⁃128⁃3p mimics on SW620 cells. Theluciferase reporter assay confirmed that miR⁃128⁃3p directly targeted to regulated ZEB1. Over ⁃ expression of ZEB1 could significantly promote the invasion and migration of SW620 cells,whereas the collaborative transfection of miR⁃128⁃3p mimics could down⁃regulate the expression of ZEB1 protein and mRNA,and reduce the invasion and migration ability of SW620 cells. Conclusions The low expression of miR⁃128⁃3pwas correlated with tumor differentiation,lymph node metastasis and distant metastasis of CRC patients. miR⁃128⁃3p could inhibit the invasion and migration of CRC cells by targeting ZEB1

Key words:

colorectal cancer, miR?128?3p, ZEB1, invasion, migration