Upregulation of miRNA‑301a‑3p promotes tumor progression in gastric cancer by suppressing NKRF and activating NF‑κB signaling

Abstract

MicroRNA-301a (miRNA/miR-301a) and nuclear factor (NF)-κB signaling play important roles in tumor invasion, migration and progression. However, the role of miRNA-301a-3p in human gastric cancer (GC), and specifically in the activation of NF-κB signaling, remains unclear. The aim of the present study was to investigate miRNA-301a-3p expression in GC progression and the molecular mechanisms as regards the regulation of NF-κB signaling. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect miRNA-301a-3p expression in GC and paired normal tissues. The association between the expression of miRNA-301a-3p and patient pathological parameters and the prognosis of GC was statistically analyzed using an in situ hybridization (ISH) assay. An MTS assay and a Transwell assay were performed to evaluate the effects of miRNA-301a-3p on the proliferation, invasion and migration of GC cells. RT-qPCR and western blot analysis were used to analyze the association between miRNA-301a-3p and nuclear factor-κB repressing factor (NKRF) expression and the corresponding downstream NF-κB signaling molecules. A luciferase assay was used to verify the target effect of miRNA-301a-3p and NKRF. It was found that miRNA-301a-3p expression was significantly higher in 30 cases of primary GC compared with matched normal tissues. Additionally, the ISH assay indicated that the high expression of miRNA-301a-3p in GC was associated with tumor invasion depth, lymph node metastasis, lymph node invasion and tumor metastasis stage. Patients whose tumors had a higher miRNA-301a-3p expression level exhibited a poorer prognosis. The in vitro assay indicated that miRNA-301a-3p affected the proliferative and invasive ability of GC cells by targeting the expression of NKRF, which then affected NF-κB signaling. Therefore, it was hypothesize that miRNA-301a-3p promotes GC progression and affects the prognosis of patients with GC by targeting NKRF, which in turn, directly influences NF-κB activation.

Signaling pathways and therapeutic interventions in gastric cancer

“…The upregulation of the NF-κB signaling pathway is involved not only in the occurrence of tumors but is also associated with chemoresistance and radioresistance. 332 , 333 NF-κB inhibitors may enhance the efficacy of antitumor drugs or increase sensitivity. With the improvement of the rapid detection technology of NF-κB activity and the understanding of the mechanism of NF-κB activation, many drugs that inhibit the activation of NF-κB have been developed.…”

PBRM1 presents a potential prognostic marker and therapeutic target in duodenal papillary carcinoma

“…The method refers to our previous published papers. 18 , 19 For detailed information on primary and secondary antibodies, please see Method S1 .…”

MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8+ T lymphocytes in human diffuse large B cell lymphoma

“…Slides were first scanned at low magnification (10x magnification), and 10 high magnification fields (400x magnification) were assessed. For scoring gene and miRNA expression, the intensity of staining was classified into 0 (no expression), 1 (weak expression), 2 (moderate expression) and 3 (high expression), and the percentage of positive cells was categorized as 1 (positive cells ≤25%), 2 (25% < positive cells ≤50%), 3 (50% < positive cells ≤75%) and 4 (positive tumor cells > 75%) [ 18 , 19 ]. The histochemical score (H-score) was achieved by multiplying the staining intensity and the percentage of positive cells and ranged from 0 to 300.…”