This study developed a novel synthetic biomimetic ligand that replicates Epidermal Growth Factor (EGF) using an integrated in silico and in vitro pipeline. A fragment-based design strategy generated a virtual library, which was filtered to identify drug-like compounds with high predicted affinity for the EGFR. The lead compound, DB09210, demonstrated exceptional biomimetic properties. In human fibroblasts, it sustained cell viability comparably to fetal bovine serum. Notably, in 3D cultures of gastric cancer stem cells (GCSCs), DB09210 exhibited potent, time- and dose-dependent activity, surpassing native EGF in promoting proliferation after 7 days. The observed maintenance of CD133 + expression can be interpreted as evidence for preserved stemness, implying the compound may act as a potent functional mimetic mechanism. While its potent enrichment of cancer stem cells requires therapeutic caution, DB09210 is a powerful tool compound. Its ability to reliably expand stem cell populations under defined, serum-free conditions holds significant promise for regenerative medicine, organoid biobanking, and advanced ex vivo drug screening platforms.
Non-small cell lung cancer (NSCLC) is a leading cause of global cancer mortality. Maoyu extract (MY) is a traditional Chinese medicine formulation comprising Ranunculus ternatus Thunb. and Houttuynia cordata Thunb., used traditionally to treat inflammatory lung ailments and nodules. This study aims to systematically investigate the anti-NSCLC efficacy of Maoyu extract and elucidate its underlying molecular mechanisms, specifically focusing on the PI3K/AKT signaling axis. Active compounds and targets were screened using the TCMSP database and validated via molecular docking. Predicted mechanisms were verified in vitro using A549 and NCI-H1299 cell lines and in vivo using a syngeneic Lewis lung carcinoma (LLC) mouse model. Network analysis identified 15 active compounds (including β-sitosterol, kaempferol, and quercetin) and 197 targets. MY significantly inhibited cell proliferation, induced apoptosis, and blocked the PI3K/AKT axis. In the LLC model, MY reduced tumor volume by 56.77
Cigarette smoke (CS) exposure is a significant environmental trigger for childhood asthma. Although S100 calcium-binding protein A10 (S100A10) is upregulated in asthma patients, its regulatory mechanism in CS-associated asthma remains unclear. This study aimed to elucidate how insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) influences S100A10 expression via the N6-methyladenosine (m6A) modification pathway and to investigate the role of the IGF2BP3/S100A10 axis in CS-exposed childhood asthma. A total of 120 asthmatic children were enrolled, with 120 healthy children selected as controls. Serum levels of S100A10 and IGF2BP3 were measured. A murine model of asthma was established using ovalbumin (OVA) induction combined with CS exposure. Lentivirus-mediated overexpression or knockdown of IGF2BP3 or S100A10 was performed to evaluate lung function, inflammatory infiltration, immune cell proportions, and immunoglobulin E (IgE) levels. Airway epithelial cells (AECs) were isolated and treated with cigarette smoke extract (CSE) and OVA to assess apoptosis and inflammatory cytokine levels. RNA immunoprecipitation, m6A methylated RNA immunoprecipitation, and crosslinking immunoprecipitation (CLIP) were conducted to validate the regulatory mechanism between IGF2BP3 and S100A10. The expression of S100A10 and IGF2BP3 was elevated in asthmatic children and OVA-induced asthmatic mice. In OVA-induced asthmatic mice, CS exacerbated OVA-induced asthma phenotypes, including increased lung resistance, enhanced inflammatory infiltration, elevated T helper 2 (Th2) and T helper 17 (Th17) cell proportions, and raised IgE levels. Knockdown of S100A10 or IGF2BP3 ameliorated these pathological changes, while overexpression of S100A10 reversed the protective effects of IGF2BP3 knockdown. Mechanistically, IGF2BP3 bound to m6A modification sites on S100A10 mRNA and enhanced its stability. CSE and OVA promoted IGF2BP3 expression, thereby increasing S100A10 mRNA stability, leading to enhanced epithelial apoptosis and release of inflammatory factors (interleukin-1 beta, interleukin-6, tumor necrosis factor-alpha). CS exposure aggravates childhood asthma by promoting IGF2BP3 expression and enhancing m6A-dependent stabilization of S100A10 mRNA. Targeting the IGF2BP3/S100A10 axis may provide a novel therapeutic strategy for CS-associated asthma.
Head and neck squamous cell carcinoma (HNSCC) is a highly heterogeneous malignancy characterized by extensive molecular dysregulation, including aberrant microRNA (miRNA) expression. Identification of clinically relevant miRNAs and their associated regulatory networks may improve understanding of oral squamous cell carcinoma (OSCC) biology and facilitate biomarker discovery. The present study aimed to identify novel dysregulated miRNAs associated with OSCC and to investigate their potential diagnostic relevance, predicted target interactions, and functional pathway associations through integrative transcriptomic and clinical validation analyses. Mature miRNA expression profiles from The Cancer Genome Atlas (TCGA)-HNSCC cohort were retrieved through the UCSC Xena Browser and analyzed using DESeq2 following anatomical site-based filtering to obtain HPV-negative OSCC samples. Differentially expressed miRNAs were identified using adjusted p-value < 0.05 and |log2FoldChange|≥ 0.5 thresholds. Based on differential expression patterns and limited prior characterization in OSCC, hsa-miR-1277, hsa-miR-301b, and hsa-miR-519a-1 were selected for downstream analyses. Kaplan–Meier survival, receiver operating characteristic (ROC), integrative target prediction, Pearson correlation, STRING protein–protein interaction network, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were subsequently performed. Independent validation was carried out using four Gene Expression Omnibus (GEO) cohorts and qRT-PCR analysis in Indian OSCC clinical specimens. Differential expression analysis identified 283 significantly expressed miRNAs in OSCC, including 167 upregulated and 116 downregulated transcripts. ROC analysis demonstrated strong discriminatory performance for hsa-miR-301b (Area Under the Curve = 0.868) and moderate performance for hsa-miR-1277 (AUC = 0.735), whereas hsa-miR-519a-1 exhibited comparatively limited diagnostic utility. Integrative target prediction identified five high-confidence targets each for hsa-miR-1277 and hsa-miR-301b, while hsa-miR-519a-1 demonstrated a broader overlap profile comprising 24 candidate genes. Functional enrichment analyses demonstrated enrichment of neuronal signaling, synaptic regulation, ion channel activity, neuroactive ligand–receptor interaction, and cadherin-associated pathways among candidate target genes. Validation across independent GEO cohorts and clinical specimens demonstrated heterogeneous expression patterns with partial concordance to TCGA-derived observations. This integrative multi-cohort analysis identifies hsa-miR-1277, hsa-miR-301b, and hsa-miR-519a-1 as novel miRNA candidates associated with OSCC-related molecular dysregulation. The findings further highlight the substantial biological heterogeneity underlying HNSCC and emphasize the challenges associated with translating large-scale transcriptomic signatures into reproducible clinical biomarkers.
Qingjin Huazhuo Formula (QJHZF) is a traditional Chinese medicinal prescription used to treat chronic obstructive pulmonary disease (COPD). To investigate its mechanism of action, a COPD rat model with phlegm heat and blood stasis syndrome was established by combining cigarette smoke exposure and passive swimming. After model confirmation, QJHZF was intragastrically administered at 10.9 g/kg/day for 14 days. The serum inflammatory cytokines and pulmonary oxidative stress indicators in rats were measured using ELISA and biochemical assays, respectively. Liquid chromatography-mass spectrometry was performed for proteomic analysis of lung tissues and identification of QJHZF components. Network pharmacology predicted potential targets for COPD treatment. After identifying the core targets through intersection analysis of proteomic targets and network pharmacology targets, qRT-PCR and molecular docking were subsequently employed for validation. Results indicate that QJHZF can inhibit inflammation and reduce oxidative stress associated with COPD. Overall, 114 differentially expressed proteins were identified. In QJHZF, 160 active compounds were identified, with 2,178 possible QJHZF targets involved in COPD treatment. A total of 29 compounds targeting 28 core targets were ultimately identified as the key constituents of QJHZF in treating COPD. qRT-PCR and molecular docking confirmed their pivotal role. KEGG enrichment analysis showed significant enrichment of the core targets in 15 pathways implicated in COPD-related inflammation and oxidative stress. Therefore, this study provides preliminary evidence for a multi-level network of 29 components − 28 targets − 15 pathways through which QJHZF treats COPD, laying a valuable foundation for further in-depth investigation.
