MiR-144-3p inhibits gastric cancer progression and stemness via directly targeting GLI2 involved in hedgehog pathway

Yixun Lu,Benlong Zhang,Wenquan Liang,Baohua Wang,Chuang Wang,Hongqing Xi,Lin Chen,Yunhe Gao,Di Wu,Xinxin Wang

doi:10.1186/s12967-021-03093-w

Abstract

BackgroundGastric cancer (GC) is the fifth most commonly diagnosed cancer worldwide. Due to the dismal prognosis, identifying novel therapeutic targets in GC is urgently needed. Evidences have shown that miRNAs played critical roles in the regulation of tumor initiation and progression. GLI family zinc finger 2 (GLI2) has been reported to be up-regulated and facilitate cancer progression in multiple malignancies. In this study, we focused on identifying GLI2-targeted miRNAs and clarifying the underlying mechanism in GC.MethodsPaired fresh gastric cancer tissues were collected from gastrectomy patients. GLI2 and miRNAs expression were detected in gastric cancer tissues and cell lines. Bioinformatics analysis was used to predict GLI2-targeted miRNAs and dual-luciferase reporter assay was applied for target verification. CCK-8, clone formation, transwell and flow cytometry were carried out to determine the proliferation, migration, invasion and cell cycle of gastric cancer cells. Tumorsphere formation assay and flow cytometry were performed to detail the stemness of gastric cancer stem cells (GCSCs). Xenograft models in nude mice were established to investigate the role of the miR-144-3p in vivo.ResultsGLI2 was frequently upregulated in GC and indicated a poor survival. Meanwhile, miR-144-3p was downregulated and negatively correlated with GLI2 in GC. GLI2 was a direct target gene of miR-144-3p. MiR-144-3p overexpression inhibited proliferation, migration and invasion of gastric cancer cells. Enhanced miR-144-3p expression inhibited tumorsphere formation and CD44 expression of GCSCs. Restoration of GLI2 expression partly reversed the suppressive effect of miR-144-3p. Xenograft assay showed that miR-144-3p could inhibit the tumorigenesis of GC in vivo.ConclusionsMiR-144-3p was downregulated and served as an essential tumor suppressor in GC. Mechanistically, miR-144-3p inhibited gastric cancer progression and stemness by, at least in part, regulating GLI2 expression.

Highlights

Gastric cancer (GC) is the fifth most commonly diagnosed cancer worldwide
GLI family zinc finger 2 (GLI2) is upregulated in gastric cancer and indicates poor survival According to recent studies, GLI2 is up-regulated in multiple cancers [9,10,11]
Through Quantitative reverse transcription PCR (qRT-PCR), we found that GLI2 was up-regulated in most gastric cancer cell lines compared with GES-1 (Fig. 1a)

Summary

Introduction

Due to the dismal prognosis, identifying novel therapeutic targets in GC is urgently needed. Evidences have shown that miRNAs played critical roles in the regulation of tumor initiation and progression. We focused on identifying GLI2-targeted miRNAs and clarifying the underlying mechanism in GC. Gastric cancer (GC) is the fifth most commonly diagnosed cancer and the fourth leading cause of cancer-related death in the world [1]. Identifying novel therapeutic targets and clarifying the underlying mechanisms are urgently needed in GC. Mountain studies have shown that miRNAs played critical roles in tumor proliferation, apoptosis, invasion, metastasis and drug resistance [5], etc. MiR203 inhibits proliferation, migration and invasion of gastric cancer cells by downregulating Slug [6]; miR-1244 inhibits tumor metastasis by targeting FAK in intestinaltype gastric cancer [7]; miR-200c functions as a tumor suppressor in gastric cancer and overcomes drug resistance during cisplatin chemotherapy [8]

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Results

Discussion

Conclusion