SUFU mediates EMT and Wnt/β-catenin signaling pathway activation promoted by miRNA-324-5p in human gastric cancer

Yin Peng; Xiaojing Zhang; Huijuan Lin; Shiqi Deng; Ying Qin; Yuan Yuan; Xianling Feng; Jian Wang; Wangchun Chen; Fan Hu; Ruibin Yan; Yanqiu Zhao; Yulan Cheng; Yanjie Wei; Xinmin Fan; Hassan Ashktorab; Duane Smoot; Song Li; Stephen J Meltzer; Zhe Jin

doi:10.1080/15384101.2020.1826632

SUFU mediates EMT and Wnt/β-catenin signaling pathway activation promoted by miRNA-324-5p in human gastric cancer

Cell Cycle. 2020 Oct;19(20):2720-2733. doi: 10.1080/15384101.2020.1826632. Epub 2020 Oct 5.

Authors

Yin Peng¹, Xiaojing Zhang^{1

2}, Huijuan Lin^{3

4}, Shiqi Deng¹, Ying Qin⁵, Yuan Yuan¹, Xianling Feng¹, Jian Wang³, Wangchun Chen¹, Fan Hu¹, Ruibin Yan⁶, Yanqiu Zhao⁶, Yulan Cheng⁷, Yanjie Wei⁸, Xinmin Fan¹, Hassan Ashktorab⁹, Duane Smoot¹⁰, Song Li⁶, Stephen J Meltzer⁷, Zhe Jin¹

Affiliations

¹ Guangdong Key Laboratory for Genome Stability & Disease Prevention, Guangdong Provincial Key Laboratory of Regional Immunity and Diseases, Department of Pathology, Shenzhen University School of Medicine , Shenzhen, Guangdong, China.
² Department of Pathology, Guangdong Province Key Laboratory of Molecular Oncologic Pathology , Guangzhou, Guangdong, China.
³ Department of Pathology and Pathophysiology, Guangzhou Medical University , Guangzhou, Guangdong, China.
⁴ Department of Ultrasound, Guangdong Women and Children Hospital , Guangzhou, Guangdong, China.
⁵ Department of Gastrointestinal Surgery, Shenzhen Second People's Hospital , Shenzhen, Guangdong, China.
⁶ Laboratory of Chemical Genomics, Shenzhen Graduate School of Peking University , Shenzhen, Guangdong, P.R. China.
⁷ Department of Medicine/GI Division, Johns Hopkins University School of Medicine and Sidney Kimmel Comprehensive Cancer Center , Baltimore, MD, USA.
⁸ Center for High Performance Computing, Shenzhen Institutes of Advanced Technology , Shenzhen, Guangdong, P.R. China.
⁹ Department of Medicine and Cancer Center, Howard University, College of Medicine , Washington, USA.
¹⁰ Department of Medicine, Meharry Medical Center , Nashville, TN, USA.

Abstract

The poor prognosis of late gastric carcinomas (GC) underscores the necessity to identify novel biomarkers for earlier diagnosis and effective therapeutic targets. MiRNA-324-5p has been shown to be over-expressed in GC, however the biological function of miRNA-324-5p implicated in gastric cancer and its downstream targets were not well understood. Wnt/β-catenin signaling pathway is aberrantly regulated in GC. We sought to explore if miRNA-324-5p promotes oncogenesis through modulating Wnt signaling and EMT. MiRNA-324-5p is highly expressed in GC based on qRT-PCR and TCGA data. In addition, in vitro cell proliferation, cell migration assays and in vivo animal exenograft were executed to show that miRNA-324-5p is an oncogenic miRNA in GC. MiRNA-324-5p activates Wnt signaling and induces EMT in GC. Further, SUFU was identified as a target of miRNA-324-5p confirmed by western blotting and luciferase assays. Spearson analysis and TCGA data indicate that the expression of SUFU is negatively associated with the expression of miRNA-324-5p. Rescue experiments were performed to determine if SUFU mediates the Wnt activation, EMT and oncogenic function of miRNA-324-5p. MiRNA-324-5p inhibitors plus SUFU siRNAs rescue partially the inhibitory effect on Wnt signaling and EMT caused by miRNA-324-5p inhibitors. Finally, the suppression of cell proliferation, migration, and colony formation ability induced by miRNA-324-5p inhibitors is alleviated by addition of SUFU siRNAs. In summary, miRNA-324-5p is overexpressed in vivo and exerts cell growth and migration-promoting effects through activating Wnt signaling and EMT by targeting SUFU in GC. It represents a potential miRNA with an oncogenic role in human gastric cancer.

Keywords: EMT; SUFU negative regulator of hedgehog signaling; Wnt/β-catenin; miRNA-324-5p.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carcinogenesis / genetics
Carcinogenesis / pathology
Cell Line
Cell Line, Tumor
Cell Movement / genetics
Cell Proliferation / genetics
Epithelial-Mesenchymal Transition / genetics
Female
Gene Expression Regulation, Neoplastic / genetics
Humans
Mice
Mice, Inbred BALB C
Mice, Nude
Mice, SCID
MicroRNAs / genetics*
Oncogenes / genetics
Organic Cation Transport Proteins / genetics*
Repressor Proteins / genetics*
Stomach Neoplasms / genetics*
Stomach Neoplasms / pathology
Wnt Signaling Pathway / genetics*
beta Catenin / genetics*

Substances

CTNNB1 protein, human
MIRN324 microRNA, human
MicroRNAs
Organic Cation Transport Proteins
Repressor Proteins
SUFU protein, human
beta Catenin
solute carrier family 22 (organic cation transporter), member 3

Grants and funding

This research is supported by: National Natural Youth Science Foundation of China (31601028) to Y Peng; National Nature Science Foundation of China (81772592) to Z Jin; National Science Foundation of China (81871969) to X Zhang; Shenzhen Basic Research Fund (JCYJ20170818142852491) to Z Jin; Nature Science Foundation of Guangdong Province (2017A030313144), Shenzhen Basic Research Fund (JCYJ20190808163801777), Medical science and technology research foundation of Guangdong Province (A2019211) and Startup Fund of Shenzhen University (2018015)to Y Peng; Medical science and technology research foundation of Guangdong Province (A2016112), Nature Science Foundation of Guangdong Province (2017A030313479) to X Zhang; National Key Research and Development Program of China (2016YFB0201305), Shenzhen Basic Research Fund (JCYJ20160331190123578, JCYJ20170413093358429) to Y Wei; Science and Technology Program of Guangdong Province (2017B030301016) to Z Jin; Medical science and technology research foundation of Guangdong Province (A2018170) to X Feng; High Quality University Construction 2nd phase (860-00000210).