长链非编码RNA SPRY4-IT1作为miR-101-5p的海绵，通过抑制ZEB1促进胃肠道间质瘤进展。

Long noncoding RNA SPRY4-IT1 acts as a miR-101-5p sponge to promote gastrointestinal stromal tumor progression by inhibiting ZEB1.

作者信息

Huang Chen, Wang Ming, Zhao Wen-Yi, Shen Yan-Ying, Zhuang Chun, Ni Bo, Yang Lin-Xi, Lu Lu, Li Xiao-Qi, Tu Lin, Cao Hui

机构信息

Department of Gastrointestinal Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University Shanghai, P. R. China.

Department of Pathology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University Shanghai, P. R. China.

出版信息

Am J Transl Res. 2023 Feb 15;15(2):1026-1040. eCollection 2023.

PMID:36915750

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10006756/

Abstract

OBJECTIVES

Research on long noncoding RNAs (lncRNAs) has been conducted in different areas of oncology. Currently, the biological significance of lncRNAs and their regulatory features in gastrointestinal stromal tumors (GIST) remain largely unknown. We have previously identified SPRY4-IT1 overexpression in GIST through lncRNA sequencing of GIST tissues. Coincidentally, SPRY4-IT1 is an intron of the SPRY4 gene, and SPRY4 is specifically highly expressed in GIST. Thus the aim of the present study was to investigate the role of lncRNA SPRY4-IT1 in GIST pathogenesis.

METHODS

Herein, we screened for SPRY4-IT1 and analyzed its possible phenotypes using Gene set enrichment analysis (GSEA). The phenotypes of GIST were verified using CCK-8, colony formation, and wound-healing assays. The ceRNA mechanism was determined by the location of lncRNA SPRY4-IT1, and its relationship to the Ago2 protein. The SPRY4-IT1/miR-101-5p/ZEB1 axis was predicted using online software and sequencing. Luciferase and pull-down assays were performed for verification. Pathway-associated and phenotype-associated proteins were detected by western blotting.

RESULTS

Sequencing analysis revealed 117 differentially expressed lncRNAs in GIST and normal gastric tissue samples. Accordingly, SPRY4-IT1 was screened out and its phenotype was predicted by GSEA. Mechanistically, SPRY4-IT1 was identified as a competing endogenous RNA (ceRNA) that downregulated miR-101-5p and upregulated ZEB1, which activated extracellular signal-regulated kinase (ERK) signaling to stimulate GIST proliferation, invasion, and epithelial-mesenchymal transition. Although this effect was regulated by a negative feedback loop through SPRY4, it was still controlled by SPRY4-IT1.

CONCLUSIONS

In GIST, we revealed a ceRNA mechanism by which SPRY4-IT1 modulates ZEB1 by sponging miR-101-5p, eventually driving tumor cell proliferation, migration, and epithelial-mesenchymal transition (EMT).

摘要

目的

长链非编码RNA（lncRNA）的研究已在肿瘤学的不同领域展开。目前，lncRNA在胃肠道间质瘤（GIST）中的生物学意义及其调控特征仍 largely未知。我们之前通过对GIST组织进行lncRNA测序，在GIST中鉴定出SPRY4-IT1过表达。巧合的是，SPRY4-IT1是SPRY4基因的一个内含子，且SPRY4在GIST中特异性高表达。因此，本研究的目的是探讨lncRNA SPRY4-IT1在GIST发病机制中的作用。

方法

在此，我们筛选出SPRY4-IT1，并使用基因集富集分析（GSEA）分析其可能的表型。GIST的表型通过CCK-8、集落形成和伤口愈合试验进行验证。ceRNA机制通过lncRNA SPRY4-IT1的定位及其与AGO2蛋白的关系来确定。使用在线软件和测序预测SPRY4-IT1/miR-101-5p/ZEB1轴。进行荧光素酶和下拉试验进行验证。通过蛋白质印迹法检测与通路相关和与表型相关的蛋白质。

结果

测序分析显示GIST和正常胃组织样本中有117种差异表达的lncRNA。据此，筛选出SPRY4-IT1，并通过GSEA预测其表型。从机制上讲，SPRY4-IT1被鉴定为一种竞争性内源RNA（ceRNA），它下调miR-101-5p并上调ZEB1，从而激活细胞外信号调节激酶（ERK）信号传导，刺激GIST增殖、侵袭和上皮-间质转化。尽管这种效应通过SPRY4受到负反馈回路的调节，但它仍然受SPRY4-IT1控制。