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

Abstract

Analysis of microRNA (miRNA) expression signatures in head and neck squamous cell carcinoma (HNSCC) has revealed that the miR-30 family is frequently downregulated in cancer tissues. The Cancer Genome Atlas (TCGA) database confirms that all members of the miR-30 family (except miR-30c-5p) are downregulated in HNSCC tissues. Moreover, low expression of miR-30e-5p and miR-30c-1-3p significantly predicts shorter survival of HNSCC patients (p = 0.0081 and p = 0.0224, respectively). In this study, we focused on miR-30e-5p to investigate its tumor-suppressive roles and its control of oncogenic genes in HNSCC cells. Transient expression of miR-30e-5p significantly attenuated cancer cell migration and invasive abilities in HNSCC cells. Nine genes (DDIT4, FOXD1, FXR1, FZD2, HMGB3, MINPP1, PAWR, PFN2, and RTN4R) were identified as putative targets of miR-30e-5p control. Their expression levels significantly predicted shorter survival of HNSCC patients (p < 0.05). Among those targets, FOXD1 expression appeared to be an independent factor predicting patient survival according to multivariate Cox regression analysis (p = 0.049). Knockdown assays using siRNAs corresponding to FOXD1 showed that malignant phenotypes (e.g., cell proliferation, migration, and invasive abilities) of HNSCC cells were significantly suppressed. Overexpression of FOXD1 was confirmed by immunostaining of HNSCC clinical specimens. Our miRNA-based approach is an effective strategy for the identification of prognostic markers and therapeutic target molecules in HNSCC. Moreover, these findings led to insights into the molecular pathogenesis of HNSCC.

Tumor-Suppressive and Immunomodulating Activity of miR-30a-3p and miR-30e-3p in HNSCC Cells and Tumoroids

“…However, the passenger strands of miR-30a and miR-30e still remain poorly studied in HNSCC. Using TCGA-HNSC data, Minemura and colleagues confirmed that miR-30a-3p and miR-30e-3p were downregulated in HNSCC [ 12 ]. Patients with low expression of miR-30e-3p had significantly poorer prognosis compared to those with a high expression [ 26 ].…”

“…Over the past decade, several studies have shown that dysregulation of miRNAs can profoundly impact normal physiology and enhance oncogenesis [ 10 , 11 ]. Analysis of miRNA signature and TCGA has revealed that the miR-30 family is frequently downregulated in HNSCC [ 12 , 13 ]. The miR-30 family contains six miRNAs (miR-30a/b/c1/c2/d/e) encoded by six genes located on three distinct loci of the human genome (chromosome 1 for miR-30e/c1, chromosome 6 for miR-30a/c2, and chromosome 8 for miR-30d/b) [ 14 ].…”

“…If guide strands of the miR-30 family share the same SEED (mRNA-targeting) sequence (5′-GUAAACA-3′), passenger strands have two different SEED sequences: 5′-UUUCAGU-3′ for miR-30a/d/e and 5′-UGGGAG-3′ for miR-30b/c. Within this family, miR-30a and miR-30e have been described as important anti-oncomiR (or tumor suppressor miRNAs) [ 12 , 13 , 15 , 16 ]. Although both miRNAs have been reported to be consistently downregulated in HNSCC compared to adjacent normal tissue [ 12 , 13 , 17 , 18 , 19 , 20 ], no data are available on the impact and consequences of variability in their expression on LA-HNSCC tumor behavior and progression.…”

“…Within this family, miR-30a and miR-30e have been described as important anti-oncomiR (or tumor suppressor miRNAs) [ 12 , 13 , 15 , 16 ]. Although both miRNAs have been reported to be consistently downregulated in HNSCC compared to adjacent normal tissue [ 12 , 13 , 17 , 18 , 19 , 20 ], no data are available on the impact and consequences of variability in their expression on LA-HNSCC tumor behavior and progression. Moreover, functional studies have focused mainly on the 5p guide strand and less on the 3p strand, which was thought to be only passenger and always eliminated.…”

Molecular Pathogenesis of Colorectal Cancer: Impact of Oncogenic Targets Regulated by Tumor Suppressive miR-139-3p

“…The assay for RIP was performed according to previous studies [ 55 ]. Briefly, CRC cells were cultured in 6-well dish at 3.0 × 10 5 /well concentration.…”