Identification of Tumor Suppressive Genes Regulated by miR-31-5p and miR-31-3p in Head and Neck Squamous Cell Carcinoma

Abstract

We identified the microRNA (miRNA) expression signature of head and neck squamous cell carcinoma (HNSCC) tissues by RNA sequencing, in which 168 miRNAs were significantly upregulated, including both strands of the miR-31 duplex (miR-31-5p and miR-31-3p). The aims of this study were to identify networks of tumor suppressor genes regulated by miR-31-5p and miR-31-3p in HNSCC cells. Our functional assays showed that inhibition of miR-31-5p and miR-31-3p attenuated cancer cell malignant phenotypes (cell proliferation, migration, and invasion), suggesting that they had oncogenic potential in HNSCC cells. Our in silico analysis revealed 146 genes regulated by miR-31 in HNSCC cells. Among these targets, the low expression of seven genes (miR-31-5p targets: CACNB2 and IL34; miR-31-3p targets: CGNL1, CNTN3, GAS7, HOPX, and PBX1) was closely associated with poor prognosis in HNSCC. According to multivariate Cox regression analyses, the expression levels of five of those genes (CACNB2: p = 0.0189; IL34: p = 0.0425; CGNL1: p = 0.0014; CNTN3: p = 0.0304; and GAS7: p = 0.0412) were independent prognostic factors in patients with HNSCC. Our miRNA signature and miRNA-based approach will provide new insights into the molecular pathogenesis of HNSCC.

MicroRNAs: Their Role in Metabolism, Tumor Microenvironment, and Therapeutic Implications in Head and Neck Squamous Cell Carcinoma

“…Upregulation of miR-31-5p has been observed in HNSCC cancers and has been confirmed by The Cancer Genome Atlas (TCGA) data analysis [63]. Functional assays have shown that miR-31-5p is an oncogenic miRNA in HNSCC [64].…”

Molecular Pathogenesis of the Coronin Family: CORO2A Facilitates Migration and Invasion Abilities in Oral Squamous Cell Carcinoma

“…Tumor-suppressive miRNAs are frequently downregulated, whereas their oncogene targets are upregulated, in cancer cells. A previous analysis of our RNA-sequencing-based miRNA signature of OSCC revealed that miR-125b-5p and miR-140-5p were downregulated in cancer tissues [ 42 ]. Our present study showed that these miRNAs have tumor-suppressive roles, and miR-125b-5p and miR-140-5p directly regulate the expression of CORO2A in OSCC cells.…”

“…RNA was extracted from cell lines and subjected to this study as described previously [ 42 , 57 , 58 ]. The TaqMan probes and primers used in this study are listed in Table S4 .…”

“…Transfection of siRNAs and miRNAs into OSCC cell lines was performed using Lipofectamine RNAiMAX reagent (Invitrogen, Carlsbad, CA, USA) according to our previous studies [ 42 , 57 , 58 ]. The reagents used in this study are listed in Table S4 .…”

“…The XTT assay for cell proliferation and the Matrigel chamber assay for the invasion were performed in OSCC cells as described previously [ 42 , 57 , 58 ]. In the wound healing assay for migration, a wound was created using a micropipette tip in the middle of each plate after siRNA or pre-miRNAs transfection 48 h. We incubated plates at 37 °C at 5% carbon dioxide with a free-serum medium, and live-cell migration was captured after 12 and 24 h.…”