Long Noncoding RNA RC3H2 Facilitates Cell Proliferation and Invasion by Targeting MicroRNA-101-3p/EZH2 Axis in OSCC

Kun Wu; Yingying Jiang; Wenkai Zhou; Bolin Zhang; Yan Li; Fei Xie; Jianjun Zhang; Xu Wang; Ming Yan; Qin Xu; Zhenhu Ren; Wantao Chen; Wei Cao

doi:10.1016/j.omtn.2020.02.006

Long Noncoding RNA RC3H2 Facilitates Cell Proliferation and Invasion by Targeting MicroRNA-101-3p/EZH2 Axis in OSCC

Mol Ther Nucleic Acids. 2020 Jun 5:20:97-110. doi: 10.1016/j.omtn.2020.02.006. Epub 2020 Feb 13.

Authors

Kun Wu¹, Yingying Jiang², Wenkai Zhou³, Bolin Zhang¹, Yan Li¹, Fei Xie¹, Jianjun Zhang³, Xu Wang³, Ming Yan³, Qin Xu³, Zhenhu Ren⁴, Wantao Chen⁵, Wei Cao⁶

Affiliations

¹ Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
² Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, School of Medicine, Shanghai Jiao Tong University, Shanghai, China; Department of Dentistry, Affiliated Hospital, Weifang Medical University, Weifang 261031, China.
³ Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China.
⁴ Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China. Electronic address: ren.zhenhu@outlook.com.
⁵ Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. Electronic address: chenwantao196323@sjtu.edu.cn.
⁶ Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. Electronic address: caowei561521@shsmu.edu.cn.

Abstract

In our previous studies, enhancer of zeste homolog 2 (EZH2) has been proven to be a key oncogenic driver in oral squamous cell carcinoma (OSCC). However, the regulatory mechanisms on EZH2 remain poorly understood in OSCC. Here, through multi-transcriptomics, bioinformatics analysis, and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the co-expression network of long noncoding RNA RC3H2 (RC3H2), microRNA-101-3p (miR-101-3p), and EZH2 were screened and validated as a competing endogenous RNA (ceRNA) mechanism in OSCC. Silencing of RC3H2 inhibited OSCC cell proliferation, colony formation, migration, and invasion in vitro and reduced the expression of EZH2 and H3K27Me3, whereas RC3H2 overexpression significantly promoted OSCC cell growth, colony formation, migration, invasion, and xenograft tumor growth in vivo and increased the expression of EZH2 and H3K27Me3. A fluorescence in situ hybridization (FISH) assay verified that RC3H2 was predominately localized to the cytoplasm. RNA pull-down and luciferase activity assays showed that miR-101-3p was physically bound to RC3H2 as well as EZH2, and its inhibitor reversed the inhibitory effect of RC3H2 knockdown on progression of OSCC. Taken together, our findings demonstrate that RC3H2 as completive endogenous RNA sponging miR-101-3p targets EZH2 and facilitates OSCC cells' malignant behavior.

Keywords: EZH2; competitive endogenous RNA; invasion; lncRNA RC3H2; metastasis; microRNA-101-3p; migration; oral squamous cell carcinoma; proliferation.