长链非编码 RNA MCM3AP-AS1 通过 miR-19a-3p/FOXF2 轴抑制结直肠癌的进展。

lncRNA MCM3AP-AS1 inhibits the progression of colorectal cancer via the miR-19a-3p/FOXF2 axis.

机构信息

Medical Care Center, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China.

Department of BIN Convergence Technology and Polymer Nano Science and Technology, Chonbuk National University, Jeonju, Republic of Korea.

出版信息

J Gene Med. 2021 Mar;23(3):e3306. doi: 10.1002/jgm.3306. Epub 2021 Jan 19.

DOI:10.1002/jgm.3306

PMID:33450091

Abstract

BACKGROUND

Long non-coding RNA MCM3AP antisense RNA 1 (lncRNA MCM3AP-AS1) has a regulatory role in the development of diverse malignancies, whereas its role and mechanism in colorectal cancer (CRC) is not yet clear.

METHODS

The relative expression of MCM3AP-AS1, miR-19a-3p and forkhead box F2 (FOXF2) mRNA in 53 cases of CRC and its adjacent normal tissues, human normal colonic mucosal cells (FHC cells) and CRC cell lines was examined by a quantitative real-time polymerase chain reaction, and the changes of cell multiplication and migration were examined by the cell counting kit-8 method, EdU test, and scratch-healing test, respectively. Bioinformatics, dual-luciferase reporter gene assay and a RNA immunoprecipitation experiment were adopted to predict and verify the relationship between MCM3AP-AS1 and miR-19a-3p; bioinformatics and dual-luciferase reporter gene assay were adopted to predict and verify the relationship between miR-19a-3p and FOXF2. Western blotting was executed to examine the effects of MCM3AP-AS1 overexpression or knockdown on FOXF2 protein expression.

RESULTS

MCM3AP-AS1 expression was down-modulated in CRC, and its dysregulation was linked to unfavorable pathological characteristics. MCM3AP-AS1 significantly impeded the multiplication and migration of CRC cells. MCM3AP-AS1 was recognized as a molecular sponge to suppress miR-19a-3p expression, and FOXF2 was a target gene of miR-19a-3p. MCM3AP-AS1 positively modulated FOXF2 expression, and its biological effect was dependent the on miR-19a-3p/FOXF2 axis.

CONCLUSIONS

MCM3AP-AS1 can inhibit CRC promoting by modulating the miR-19a-3p/FOXF2 axis.

摘要

背景

长链非编码 RNA MCM3AP 反义 RNA 1（lncRNA MCM3AP-AS1）在多种恶性肿瘤的发展中具有调节作用，但其在结直肠癌（CRC）中的作用和机制尚不清楚。

方法

采用实时定量聚合酶链反应检测 53 例 CRC 及其相邻正常组织、人正常结肠黏膜细胞（FHC 细胞）和 CRC 细胞系中 MCM3AP-AS1、miR-19a-3p 和叉头框 F2（FOXF2）mRNA 的相对表达量，采用细胞计数试剂盒-8 法、EdU 试验和划痕愈合试验分别检测细胞增殖和迁移的变化。采用生物信息学、双荧光素酶报告基因检测和 RNA 免疫沉淀实验预测和验证 MCM3AP-AS1 与 miR-19a-3p 之间的关系；采用生物信息学和双荧光素酶报告基因检测预测和验证 miR-19a-3p 与 FOXF2 之间的关系。采用 Western blot 检测 MCM3AP-AS1 过表达或敲低对 FOXF2 蛋白表达的影响。

结果

CRC 中 MCM3AP-AS1 表达下调，其失调与不良的病理特征有关。MCM3AP-AS1 显著抑制 CRC 细胞的增殖和迁移。MCM3AP-AS1 被认为是一种分子海绵，可以抑制 miR-19a-3p 的表达，FOXF2 是 miR-19a-3p 的靶基因。MCM3AP-AS1 正向调节 FOXF2 表达，其生物学效应依赖于 miR-19a-3p/FOXF2 轴。