NF-κB-Induced Upregulation of miR-548as-3p Increases Invasion of NSCLC by Targeting PTEN

Abstract

Background: Non-Small Cell Lung Cancer (NSCLC) is an aggressive cancer type due to high metastatic capacity. Nuclear Factor Kappa B (NF-κB) is a consistently active transcription factor in malignant lung cancer cells and has crucial significance in NSCLC progression. It is also implicated in the transcriptional regulation of many genes including microRNAs (miRNAs) that function as tumor suppressor or oncogene. It has been increasingly reported that several miRNAs defined as gene members are induced by NF-κB. The present study aimed to find novel miRNAs that are regulated by NF-κB. Methods: Chromatin İmmunoprecipitation Sequencing (ChIP-Seq) experiment and bioinformatic analysis were used to determine NF-κB-dependent miRNAs. Western blot analysis, quantitative real-time polymerase chain reaction (qRT-PCR), luciferase reporter gene assays were carried out to investigate the target genes of miRNAs. To determine biologic activity, transwell invasion and MTT assay were carried out on H1299 NSCLC cell line. miRNA expression level was evaluated in metastatic and non-metastatic tissue samples of NSCLC patients. Results: ChIP-Seq and qRT-PCR experiments showed that miR-548as-3p is transcriptionally regulated by NF- κB in response to Tumor Necrosis Factor-α (TNF-α) treatment. Then, we found that tumor suppressor Phosphatase and Tension homolog (PTEN) is a direct target of miR-548as-3p. Furthermore, miR-548as-3p mediates phosphatidylinositol-3-OH kinase (PI3K)/Akt pathway and NF-κB-implicated genes including Matrix Metalloproteinases 9 (MMP9), Slug and Zeb1. We further showed that miR-548as-3p increased invasiveness of NSCLC cells and was upregulated in metastatic tumor tissues compared to non-metastatic ones. Conclusion: All these findings provide a miRNAs-mediated novel mechanism for NF-κB signaling and that miR-548as-3p could be a biomarker for NSCLC metastasis.

miR‑494 promotes progression of retinoblastoma via PTEN through PI3K/AKT signaling pathway

“…PTEN is essential for the maintenance of normal cells and acts as a tumor suppressor in human cancers ( 31 ). A large number of studies have reported that PTEN played important roles in tumor development and progression, including lung cancer, ovarian cancer, and prostate cancer ( 23 , 32 , 33 ). Furthermore, in the RB development, PTEN showed an inhibitory effect and it served as the target of multiple miRNAs.…”

“…For instance, miR-296/PTEN axis facilitated gastric cancer tumorigenesis in vitro and in vivo ( 22 ). miR-548 promoted NSCLC cell invasion by regulating PTEN ( 23 ). Moreover, miR-182 promoted breast cancer cell viability and invasion via targeting PTEN ( 24 ).…”

The NF‐κB‐modulated miR‐19a‐3p enhances malignancy of human ovarian cancer cells through inhibition of IGFBP‐3 expression

“…Aberrant miRNAs expression can also be regulated by NF‐κB. The expression of miR‐31‐5p, miR‐146a, and miR‐548as‐3p can be upregulated by NF‐κB, whereas miR‐195 and miR‐497 are negatively regulated with NF‐κB . It has been reported that miR‐19a promotes colitis‐associated colorectal cancer by regulating tumor necrosis factor α‐induced protein 3‐NF‐κB feedback loops and microRNA‐19a promotes cell viability and migration of chondrocytes via upregulation of SOX9 through the NF‐κB pathway .…”

Integrative Bioinformatics Analysis Reveals Potential Target Genes and TNFα Signaling Inhibition by Brazilin in Metastatic Breast Cancer Cells

“…Another study demonstrated that PTEN hampers invasion and metastasis of gastric cancer cells through the inhibition of the PI3K/NFkB pathway and preventing the DNA binding of NFkB on the FAK promoter (Zhang et al, 2014). Moreover, PTEN inhibits metastasis in breast cancer cells through the downregulation of WNT1 inducible signaling pathway protein 1 (WISP1) and lipocalin-2 (LCN2) (Chiang et al, 2016), and inactivation of NFkB signaling in non-small cell lung cancer cells (Akgun et al, 2019). A previous study showed that TNFα increased the expression of PTEN through activation of NFkB signaling in HL60 human leukemic cells (Lee et al, 2007).…”