Aberrant miR-874-3p/leptin/EGFR/c-Myc signaling contributes to nasopharyngeal carcinoma pathogenesis

Abstract

Background Leptin is important in physiological and pathological functions in various cancers, however, the significance and mechanisms of leptin in nasopharyngeal carcinoma remain ambiguous. Methods Leptin expression was analyzed by QPCR, immunohistochemistry, Western blotting, and TCGA database. The impact of gain- or loss-of-function of leptin were determined by MTT, colony formation, wound healing, and Transwell assays in NPC cells, and by a xenograft tumor model. Leptin-modulated glucose consumption and lactate production were assessed by ELISA. Furthermore, leptin-regulated signaling pathways were examined by QPCR and Western blotting assays. The immunoprecipitation assay was conducted to determine interaction between leptin and EGFR. In addition, miR-874-3p-regulated leptin expression was evaluated using bioinformatics, QPCR, luciferase assay, AGO2-RIP assay, and Western blotting. Results In this study, we found that leptin was highly expressed in the sera and tumor tissues of patients with NPC, and elevated leptin expression was associated with advanced clinical features and poor prognosis. Functional assays demonstrated that leptin remarkably promoted NPC cell growth, motility, and glycolysis in vitro and in vivo. Mechanistically, leptin associated with EGFR, resulting in enhanced cell growth through the regulation of cell-cycle related markers, glycolysis-related genes, and EGFR/AKT/c-Myc signaling. Moreover, leptin potentiated the invasive capacity of NPC cells by promoting EMT. We further explored that miR-874-3p influenced leptin-mediated NPC progression. Overexpression of miR-874-3p prevented cell growth, motility, glucose consumption, and lactate production in NPC cells, whereas miR-874-3p inhibition had the opposite effects. AGO-RIP assays confirmed that Argonaute 2 (AGO2), a protein associated with miR-874-3p, regulated leptin expression in NPC cells. The rescue assays indicated that inhibition of leptin suppressed the effects of miR-874-3p inhibitor. In clinical specimens, miR-874-3p was negatively correlated with leptin. Conclusions Leptin may serve as a novel prognostic factor and potential therapeutic target for patients with NPC. In addition, a newly discovered regulatory axis of leptin/EGFR/AKT/c-Myc can provide a novel therapeutic strategy for NPC.

Tumor-associated characteristics and immune dysregulation in nasopharyngeal carcinoma under the regulation of m7G-related tumor microenvironment cells

“…Furthermore, we found that B_C3 was enriched in the MYC targets V1/V2 (Fig. 5 D), which is linked to NPC tumorigenesis [ 31 ]. TFs promoting NPC cell proliferation, migration and invasion, and EMT progression, such as SREBF1 [ 32 ], SOX9 [ 33 ], and SOX4 [ 34 ], were significantly activated in the m7G-related B cell clusters (Fig.…”

miR-874-3p Is Identified as a Biomarker for Acute Kidney Injury and Mediates Disease Development via Targeting MSRB3

“…For instance, the downregulation of miR-874-3p in hepatocellular carcinoma indicated poorly differentiated tumor cells, advanced stages, and adverse patients' outcomes [33]. In the sera and tumor tissues of nasopharyngeal carcinoma patients, miR-874-3p was found to be upregulated, which was associated with malignant development and poor prognosis of patients, and it was also found to regulate cellular processes during tumor progression [34]. Herein, significant downregulation of miR-874-3p was observed in the serum of AKI patients, which could discriminate AKI patients from healthy individuals with relatively high sensitivity and specificity suggesting its potential in serving as a diagnostic biomarker of AKI.…”

Influence of metabolic stress and metformin on synaptic protein profile in SH‐SY5Y‐derived neurons

“…Synaptophysin (Cell Signaling #5461 (Wu et al., 2018)), NeuN (Cell Signaling #24307 (Tang et al., 2021)), PSD‐95 (post‐synaptic marker, Cell Signaling #36233 (Shui et al., 2022)), Homer‐1 (post‐synaptic marker, SC‐136358 (Wang et al., 2014)), SNAP‐25 (Cell signaling #5308 (Polishchuk et al., 2023)), VAMP2 (Cell signaling #13508 (Arrojo et al., 2019)) were measured as synaptic and neuronal markers. Total IR α (Cell signaling #74118 (Dall'Agnese et al., 2022)), total IR β (Cell signaling #23413 (Dall'Agnese et al., 2022)), IRS1 S636 (Cell signaling #2388 (Uddin et al., 2019)), total IRS1 (Cell signaling #2390 (Tang et al., 2019)), mTOR S2448 (Cell signaling #2971 (Luo et al., 2022)), total mTOR (Cell signaling #2972 (Luo et al., 2022)), Akt T308 (Cell signaling #9275 (Mathieu et al., 2019)), Akt S473 (Cell signaling #4058 (Marko et al., 2020)), and total Akt (Cell signaling #4685 (Marko et al., 2020)) were measured as markers of insulin signaling. Ponceau S, α ‐tubulin (Cell signaling #2144 (Mertins et al., 2021)), GAPDH (Abcam ab8245 (Luo et al., 2023)) or β ‐Actin (Abcam ab8227 (Moll et al., 2023)) were utilized as loading controls.…”

“…Total raptor (Cell Signaling #2280 (Sanders et al, 2022)) and phosphorylated raptor (S792, Cell Signaling #2083 (Sanders et al, 2022)) levels were also measured alongside total P70S6K (Cell Signaling #9202 (Zhang et al, 2021)), phosphorylated P70S6K (Santa Cruz SC-11759 (Xiao et al, 2002)), total ULK (Cell signaling #8054 (Wu et al, 2020)), and ULK S555 (cell signaling #5869 (Wu et al, 2020)). Synaptophysin (Cell Signaling #5461 (Wu et al, 2018)), NeuN (Cell Signaling #24307 (Tang et al, 2021)), PSD-95 (post-synaptic marker, Cell Signaling #36233 (Shui et al, 2022)), Homer-1 (post-synaptic marker, SC-136358 (Wang et al, 2014) (Tang et al, 2019)), mTOR S2448 (Cell signaling #2971 (Luo et al, 2022)), total mTOR (Cell signaling #2972 (Luo et al, 2022)), Akt T308 (Cell signaling #9275 (Mathieu et al, 2019)), Akt S473 (Cell signaling #4058 (Marko et al, 2020)), and total Akt (Cell signaling #4685 (Marko et al, 2020)) were measured as markers of insulin signaling. Ponceau S, α-tubulin (Cell signaling #2144 (Mertins et al, 2021)), GAPDH (Abcam ab8245 (Luo et al, 2023)) or β-Actin (Abcam ab8227 (Moll et al, 2023)) were utilized as loading controls.…”