长链非编码 RNA ANCR 通过调节 FoxO1 表达抑制巨噬细胞 M1 极化促进胃癌侵袭和迁移。

LncRNA ANCR Promotes Invasion and Migration of Gastric Cancer by Regulating FoxO1 Expression to Inhibit Macrophage M1 Polarization.

机构信息

Department of Oncology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China.

Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Nanchang University, No. 1 Min-De Road, Nanchang, 330006, Jiangxi Province, China.

出版信息

Dig Dis Sci. 2020 Oct;65(10):2863-2872. doi: 10.1007/s10620-019-06019-1. Epub 2020 Jan 1.

DOI:10.1007/s10620-019-06019-1

PMID:31894487

Abstract

BACKGROUND

Long non-coding RNAs (LncRNAs) are closely related to the occurrence of cancer, but its mechanism in gastric cancer (GC) is still largely unclear.

AIMS

This study aimed to reveal the underlying mechanism of LncRNA ANCR in GC.

METHODS

The expression of LncRNA ANCR was detected by qRT-PCR. ELISA was used to identify THP-1 cells into macrophage M1 type polarization. After macrophages overexpressing LncRNA ANCR were co-cultured with GC cell HGC-27, the invasion and metastasis of GC were analyzed by Transwell assay. The targeted regulation of FoxO1 by LncRNA ANCR was analyzed by RNA pull-down, RNA immunoprecipitation (RIP), and Western blot. The BALB/c nude mouse model of GC was established to analyze the effect of LncRNA ANCR on tumor growth.

RESULTS

LncRNA ANCR was highly expressed in GC. The overexpression of LncRNA ANCR in macrophages reduced the concentrations of M1 macrophage polarized marker molecules IL-1β and IL-6 in the supernatant of cells, and inhibited the polarization of macrophages to M1, while the knockdown of LncRNA ANCR produced the opposite effect. The co-culture of macrophages overexpressing LncRNA ANCR with GC cells promoted the invasion and migration of cells. LncRNA ANCR targeted FoxO1 and inhibited the expression of FoxO1 in THP-1 cells by promoting FoxO1 ubiquitination degradation. In addition, the overexpression of LncRNA ANCR promoted tumor growth in a BALB/c nude mouse model of GC, while the knockdown of LncRNA ANCR produced the opposite effect.

CONCLUSIONS

Based on these results, the overexpression of LncRNA ANCR promoted the invasion and metastasis of GC cells via down-regulating FoxO1 to inhibit macrophage polarization to M1.

摘要

背景

长链非编码 RNA（LncRNA）与癌症的发生密切相关，但在胃癌（GC）中其机制仍在很大程度上不清楚。

目的

本研究旨在揭示 LncRNA ANCR 在 GC 中的潜在机制。

方法

通过 qRT-PCR 检测 LncRNA ANCR 的表达。采用 ELISA 将 THP-1 细胞诱导为巨噬细胞 M1 型极化。在过表达 LncRNA ANCR 的巨噬细胞与 GC 细胞 HGC-27 共培养后，通过 Transwell 分析 GC 细胞的侵袭和转移。通过 RNA 下拉、RNA 免疫沉淀（RIP）和 Western blot 分析 LncRNA ANCR 对 FoxO1 的靶向调节。建立 GC 的 BALB/c 裸鼠模型，分析 LncRNA ANCR 对肿瘤生长的影响。

结果

LncRNA ANCR 在 GC 中高表达。巨噬细胞中 LncRNA ANCR 的过表达降低了细胞上清液中 M1 巨噬细胞极化标记分子 IL-1β和 IL-6 的浓度，并抑制了巨噬细胞向 M1 的极化，而 LncRNA ANCR 的敲低则产生了相反的效果。过表达 LncRNA ANCR 的巨噬细胞与 GC 细胞共培养促进了细胞的侵袭和迁移。LncRNA ANCR 通过促进 FoxO1 泛素化降解靶向 FoxO1 并抑制 THP-1 细胞中 FoxO1 的表达。此外，LncRNA ANCR 的过表达促进了 GC 的 BALB/c 裸鼠模型中的肿瘤生长，而 LncRNA ANCR 的敲低则产生了相反的效果。