Identification of Tumor-Suppressive miR-30e-3p Targets: Involvement of SERPINE1 in the Molecular Pathogenesis of Head and Neck Squamous Cell Carcinoma

Abstract

Recently, our studies revealed that some passenger strands of microRNAs (miRNAs) were closely involved in cancer pathogenesis. Analysis of miRNA expression signatures showed that the expression of miR-30e-3p (the passenger strand of pre-miR-30e) was significantly downregulated in cancer tissues. In this study, we focused on miR-30e-3p (the passenger strand of pre-miR-30e). We addressed target genes controlled by miR-30e-3p that were closely associated with the molecular pathogenesis of head and neck squamous cell carcinoma (HNSCC). Ectopic expression assays demonstrated that the expression of miR-30e-3p attenuated cancer cell malignant phenotypes (e.g., cell proliferation, migration, and invasive abilities). Our analysis of miR-30e-3p targets revealed that 11 genes (ADA, CPNE8, C14orf126, ERGIC2, HMGA2, PLS3, PSMD10, RALB, SERPINE1, SFXN1, and TMEM87B) were expressed at high levels in HNSCC patients. Moreover, they significantly predicted the short survival of HNSCC patients based on 5-year overall survival rates (p < 0.05) in The Cancer Genome Atlas (TCGA). Among these targets, SERPINE1 was found to be an independent prognostic factor for patient survival (multivariate Cox regression; hazard ratio = 1.6078, p < 0.05). Aberrant expression of SERPINE1 was observed in HNSCC clinical samples by immunohistochemical analysis. Functional assays by targeting SERPINE1 expression revealed that the malignant phenotypes (e.g., proliferation, migration, and invasion abilities) of HNSCC cells were suppressed by the silencing of SERPINE1 expression. Our miRNA-based approach will accelerate our understanding of the molecular pathogenesis of HNSCC.

Identifying Oxidized Lipid Metabolism-Related LncRNAs as Prognostic Biomarkers of Head and Neck Squamous Cell Carcinoma

“…Previous research has indicated that SERPINE1 plays a crucial role in controlling the origin and development of HNSCC and might be identified as critical indicators for the accurate diagnosis and prognosis of HNSCC; hence, providing prospective therapeutic targets [ 51 ]. Consistent with its relationship with a poor prognosis in HNSCC patients, SERPINE1 overexpression enhances tumor aggressiveness and metastasis to lymph nodes and the lung [ 52 ]. In addition, functional experiments targeting SERPINE1 expression have indicated that suppressing SERPINE1 expression inhibited the malignant phenotype of HNSCC cells [ 53 ].…”

Identification of Antitumor miR-30e-5p Controlled Genes; Diagnostic and Prognostic Biomarkers for Head and Neck Squamous Cell Carcinoma

“…The protocols used for transient transfection of miRNAs and siRNAs were described in our previous studies [ 23 , 24 , 25 , 26 ]. The miRNA precursors and siRNAs used in this report were detailed in Table S2 .…”

“…The procedures for conducting functional assays (cell proliferation, migration, and invasion assays) with HNSCC cells were described in earlier publications [ 23 , 24 , 25 , 26 ]. In brief, for proliferation assays, Sa3 or SAS cells were transferred to 96-well plates at 3.0 × 10 3 cells per well.…”

“…The procedures for plasmid construction and the dual-luciferase reporter assays were outlined previously [ 23 , 24 , 25 , 26 ]. Briefly, a partial wild-type sequence, including the seed sequence of FOXD1 within the 3′-untranslated region (3′-UTR), was inserted into the psiCHECK-2 vector (C8021; Promega, Madison, WI, USA).…”

“…The procedures for immunohistochemistry were described in our previous studies [ 23 , 24 , 25 , 26 ]. The clinical samples were obtained from HNSCC cases who received surgical treatment at Chiba University Hospital.…”