26 July 2019 : Laboratory Research
Small Nucleolar RNA Host Gene 12 (SNHG12) Promotes Proliferation and Invasion of Laryngeal Cancer Cells via Sponging miR-129-5p and Potentiating WW Domain-Containing E3 Ubiquitin Protein Ligase 1 (WWP1) ExpressionJing Li1ABCF, Suguang Sun1BCE, Wei Chen1ABCDEFG*, Kun Yuan1E
Med Sci Monit 2019; 25:5552-5560
BACKGROUND: The clinical significance and biological function of long noncoding RNA SNHG12 have not been identified in laryngeal squamous cell carcinoma (LSCC).
MATERIAL AND METHODS: Expression levels of SNHG12, miR-129-5p, and WWP1 in LSCC tissues or cells were tested by RT-qPCR. MTT assay, flow cytometry, and Transwell assay were used to identify the progression of LSCC cells in vitro. Luciferase reporter assay was used to assess the associations among SNHG12, WWP1, and miR-129-5p.
RESULTS: SNHG12 was significantly overexpressed in LSCC tissues compared with adjacent normal tissues. The expression level of SNHG12 was significantly associated with T classification, lymph node metastasis, and cancer stage of LSCC. High expression of SNHG12 predicted shorter disease-free survival. Suppressing SNHG12 using siRNA inhibited proliferation and invasion and promoted apoptosis in the AMC-HN-8 LSCC cell line. SNHG12, mainly located in cytoplasm of AMC-HN-8 cells, was validated by dual luciferase reporter test and RT-qPCR to directly interact with miR-129-5p. Inhibition of miR-129-5p significantly increased proliferation and invasion of AMC-HN-8 cells and ameliorated the suppressive effects of si-SNHG12. Luciferase assay showed that miR-129-5p was able to combine with the 3’UTR region of WWP1, which is generally regarded as an E3 ubiquitin protein ligase. RT-qPCR and Western blot showed that WWP1 was positively regulated by SNHG12 and negatively regulated by miR-129-5p at the mRNA level and protein level. Overexpression of WWP1 significantly increased proliferation and invasion of laryngeal cancer cells. Moreover, when SNHG12 was suppressed, rescue of WWP1 restored the proliferation and invasion abilities of AMC-HN-8 cells.
CONCLUSIONS: Our study demonstrated that SNHG12 promoted LSCC cells progression via sponging miR-129-5p and potentiating WWP1 expression.
Keywords: Laryngeal Neoplasms, RNA, Long Noncoding, Aged, Gene Expression, Middle Aged, Neoplasm Invasiveness, Prognosis, Squamous Cell Carcinoma of Head and Neck, transcriptome, Ubiquitin-Protein Ligases
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