MiR-124-3p impedes the metastasis of non-small cell lung cancer via extracellular exosome transport and intracellular PI3K/AKT signaling

Abstract

Background Metastasis is a significant factor that affects the survival of patients with non-small cell lung cancer (NSCLC). Nevertheless, the molecular regulatory mechanism underlying the metastasis is currently not fully understood. This study aims to identify the important role of miR-124-3p in metastasis of NSCLC, thereby providing a potential therapeutic intervention. Methods Exosome secretion was determined by Nanoparticle Tracking Analysis (NTA) and the uptake was measured by fluorescence inverted microscope. The binding mechanism between miR-124-3p and its upstream or downstream target genes was validated experimentally by Luciferase reporter. Cells migration was evaluated by transwell assays. Transcriptome sequencing on A549 was carried out to verify the potential signaling pathway underlying miR-124-3p regulation. Western blotting analysis was used to assess the level of AKT, p-AKT, PI3K, and p-PI3K protein expression in NSCLC cell lines. The role of miR-124-3p to suppress the tumor metastasis was verified in NSCLC xenograft model. Results Exosomes were more abundant in serum from patients with advanced lung cancer (n = 24 patients) than in these from patients with early-stage lung cancer (n = 30 patients), which suggested the potential correlation between amount of exosome secretion and the metastasis of NSCLC. Interestingly, the exosome release, uptake and the migration of NSCLC cells were notably inhibited by miR-124-3p. LINC00511 suppressed the expression of miR-124-3p to facilitate exosome transport due to its role as the competitive endogenous RNA for miR-124-3p. The miR-124-3p could directly target the 3′-UTR of Rab27a in NSCLC cells to inhibit exosome secretion and thereby prevent cell migration and invasion. Aside from the inhibition of exosome transport, miR-124-3p inhibited the activation of PI3K/AKT signaling in the intracellular environment. Finally, by measuring subcutaneous tumor weight and volume and lung metastasis, we also demonstrated that miR-124-3p inhibited tumor growth in vivo. Conclusion In NSCLC, miR-124-3p significantly suppressed metastasis through extracellular exosome transport and intracellular PI3K/AKT signaling. These findings provide new insights toward a better understanding of the NSCLC metastasis and suggest a potential treatment biomarker for NSCLC.

A potential posttranscriptional regulator for p60‐katanin: miR‐124‐3p

“…(δ-like canonical notch ligands delta-like 4 protein) draws attention to interaction with miR-124 in neural stem cell differentiation (Jiao et al, 2017). In other studies, miR-124-3p was shown to suppress the invasiveness and metastasis of hepatocarcinoma cells (Majid et al, 2020) and nonsmall cell lung cancer cells (Zhu et al, 2023) which indicates the role of miR-124-3p on metastasis. In addition, overexpression of miR-124-3p in human glioblastoma multiforme and stem cells derived from mouse neural brain tumors was shown to induce conversion to the neural phenotype (Silber et al, 2008).…”

miR-124 as a Liquid Biopsy Prognostic Biomarker in Small Extracellular Vesicles from NSCLC Patients

“…Patients with above-median sEV levels had a poorer prognosis, with a 2.4 times higher risk of progression and 3 times higher mortality than patients with below-median levels, in whom the risk of death was reduced by almost 70%. Furthermore, having shown that the sEV content of these patients is rich in miR-124, the fact that other authors have found an opposite prognostic risk when using the tissue expression levels of miR-124 [99], and that miR-124 can act as a negative regulator of metastasis in lung cancer through targeting the GTPases responsible for exosome export [66,100], supports our hypothesis regarding the active expulsion of tumor suppressor miRNAs by cells during carcinogenesis in lung cancer, even in early stages, to promote the progression of the disease. Conversely, we did not find any association with the prognoses of NSCLC patients when we analyzed the levels of miR-132 in sEVs, suggesting that miR-132 might not be a good candidate for monitoring advanced NSCLC progression.…”

Genome-Wide Methylation Analysis in Two Wild-Type Non-small Cell Lung Cancer Subgroups with Low and High PD-L1 Expression Levels

“…It plays a role in both breast and lung cancer [48] and was shown to suppress metastasis of lung cancer. [49] TRIM71 (Tripartite Motif Containing 71, LIN41) is a gene that encodes an E3 ubiquitin-protein ligase, which plays a role in the G1-S phase transition of the cell cycle. TRIM7 negatively regulates the NF-kappa B signaling pathway in lung cancer by degrading p65.…”