Current Medicinal Chemistry - Online First

This study examined the regulatory mechanisms and functional significance of LINC01515 in lung adenocarcinoma (LUAD) cells.

Associations between LINC01515 levels and patient survival were analyzed using TCGA data, and qRT-PCR was employed to assess LINC01515 transcript levels in both LUAD tissues (n = 15) and cell lines. Following LINC01515 silencing, changes in migration, proliferation, cellular viability, and invasion were evaluated using colony formation, CCK-8, and Transwell assays. Bioinformatics predictions of relationships among LINC01515, miR-33a-5p, and HMGA2 were confirmed using dual-luciferase reporter assays. Functional cellular experiments further verified that LINC01515 modulates both growth and motility via the miR-33a-5p/HMGA2 regulatory axis, and mouse xenograft models were employed to determine the effects of LINC01515 depletion on tumor development.

Compared with matched normal controls, LINC01515 levels were markedly higher in LUAD tissues (p<0.01). Moreover, both cellular and animal functional tests revealed that LINC01515 silencing significantly decreased cell motility, colony-forming capacity, proliferation, and invasive potential. Further mechanistic investigations showed that LINC01515 acts as a competing endogenous RNA by sequestering miR-33a-5p, thereby alleviating the suppression of HMGA2 expression. Moreover, rescue assays confirmed that the regulatory interplay among LINC01515, miR-33a-5p, and HMGA2 is a key determinant of LUAD cell proliferation and motility.

These data identify LINC01515 as an oncogenic driver of LUAD via a cytoplasmic ceRNA mechanism that derepresses HMGA2 through miR-33a-5p. Clinically, measuring LINC01515 may aid risk stratification and prognosis, and therapeutically, targeting LINC01515 or restoring the miR-33a-5p/HMGA2 balance represents a tractable axis to curb LUAD growth and dissemination, extending the lncRNA-based framework for precision oncology.

LINC01515 is predominantly distributed in the cytoplasm of LUAD cells and promotes oncogenic behavior through modulation of the miR-33a-5p/HMGA2 axis, suggesting the potential of LINC01515 as a biomarker and target in LUAD management.

The coronavirus 3C-like protease (3CLpro) is essential for SARS-CoV-2 replication, making it a key target for antiviral drug development. Natural products represent a valuable source of bioactive compounds. This study aimed to identify novel 3CLpro inhibitors from natural compounds through a combination of virtual screening and experimental validation.

Recombinant 3CLpro was expressed, purified, and evaluated for enzymatic activity using Fluorescence Resonance Energy Transfer (FRET) assays under optimized conditions. Out of 583 virtually screened compounds, 30 were selected for experimental validation. Epitheaflagallin 3-O-gallate (ETFGg) was further analyzed for binding interactions using Molecular Dynamics (MD) simulations.

ETFGg exhibited strong binding affinity (−66.90 kcal/mol) and inhibitory activity (IC50 = 8.73 ± 2.30 μM) against 3CLpro. MD simulations revealed stable interactions with key residues (HIE163, THR190, GLN192) in the 3CLpro active site.

Our findings demonstrate that ETFGg is a potent and stable inhibitor of SARS-CoV-2 3CLpro, with a binding profile distinct from known inhibitors such as ebselen. The substrate inhibition kinetics observed suggest a novel allosteric mechanism, which may provide a new strategy for targeting 3CLpro. This study supports the value of natural product libraries combined with computational and FRET-based screening for discovering antiviral leads.

ETFGg was identified as a promising 3CLpro inhibitor with high binding stability, highlighting its potential as a lead compound for the development of anti-COVID-19 drugs.

Lung cancer is a leading cause of cancer-related morbidity and mortality. The development and evaluation of effective treatment strategies for lung cancer are of high clinical importance. Everolimus (Eve) has been shown to upregulate the expression of phosphatases and inhibit the migration and proliferation of A549 cancer cells. The present study focuses on the synthesis of biodegradable bovine serum albumin (BSA) nanoparticles for the loading and delivery of Eve.

In the desolvation process, Eve molecules were kept in the BSA system. The physicochemical properties of the Eve drug containing BSA nanoparticles (Eve@BSA) have been exactly characterized. The loading and release assays of Eve were also studied at different glutaraldehyde percentages, times, and solvents.

Field emission scanning electron microscopy (FE-SEM) analysis of BSA nanoparticles revealed a spherical morphology with an average size of 93.7 ± 3.7 nm. The results demonstrated that BSA nanoparticles are highly efficient carriers, achieving an Eve loading efficiency of approximately 54% at 4% glutaraldehyde. The release of Eve from the BSA nanoparticles was dependent on the solvent and duration of incubation. According to the MTT assay, Eve@BSA exhibited low cytotoxicity and high biocompatibility against L929 fibroblast cells. In contrast, the cytotoxicity of Eve@BSA against A549 cells (IC50 ≈ 47 μg/mL) was significantly higher than that of free Eve (IC50 ≈ 283 μg/mL) after 48 hours.

The synergistic effects of Eva@BSA nanoformulation due to functional groups-rich BSA seemed to improve in vitro antiproliferation efficacies compared with the single treatment of Eve.

The findings confirm the synergistic anticancer effect of Eve@BSA, indicating that this nanosystem may serve as a promising candidate for the treatment of lung cancer.

The development and progression of non-small cell lung cancer (NSCLC) are intricately linked to immune cell activation, but its related signature has not been reported.

This study combines in silico and in vitro approaches. TCGA-NSCLC and Gene Expression Omnibus (GEO) datasets were utilized to develop and validate a prognostic signature based on cell activation genes. The signature’s validity was assessed through the identification of genomic, transcriptomic, tumor microenvironment (TME), and single-cell infiltration characteristics. The function of the candidate gene RORA was verified using CCK8, apoptosis, colony formation, wound healing, and transwell assays. The detailed mechanism of RORA was investigated through ChIP-PCR, luciferase assays, Western blot, and ROS detection.

The prognostic signature was constructed from TCGA-NSCLC datasets and validated in six independent datasets (GSE30219, GSE33072, GSE37745, GSE41271, GSE42127, GSE50081). The signature was associated with LRP1B and RYR2 mutations, NSCLC-related pathways, drug response, and immune cell infiltration. The candidate gene RORA significantly inhibits the proliferation and migration abilities of NSCLC cell lines (A549 and NCI-H1299). Furthermore, the transcription factor RORA promotes ZNF490 expression, which subsequently inhibits NUDFs expression and oxidative phosphorylation (oxphos).

The signature highlighted its significance with genomic features that were frequently reported as prognostic indicators (LRP1B and RYR2 mutations, cancer-related infiltration and pathway infiltration), and putative treatment response (IC50 in the TCGA dataset). Its detailed mechanism of candidate gene RORA revealed its role in oxphos, highlighting the crosstalk between metabolism and immune activation.

The model is robust and effectively reflects NSCLC heterogeneity while predicting prognosis. RORA promotes the expression of ZNF490 to inhibit NUDFs and oxidative phosphorylation.

An excessive inflammatory response plays a central role in the pathogenesis of ulcerative colitis (UC), but the specific cytokines involved remain unclear. This study aimed to identify inflammatory factors associated with UC and explore the possible mechanisms of the identified targets.

Protein quantitative trait loci (pQTLs) and expression quantitative trait loci (eQTLs) for inflammatory cytokines were obtained from a genome-wide pQTL study and the eQTL consortium, respectively. Summary data for UC from the exploration and validation cohorts were derived from a genome-wide association study and the Finngen cohort. MR and colocalization analyses were conducted to identify causal associations between inflammatory cytokines and UC. Bioinformatics analyses were employed to explore the involved biological processes of candidate targets. Immunohistochemistry was used to validate the expression of these candidate targets in colon tissues.

Among all inflammatory cytokines, a significant causal association was identified between C-X-C motif chemokine ligand 5 (CXCL5) and UC. Using eQTL data, a significant genetic association was established between the mRNA expression of CXCL5 and its receptor, C-X-C motif chemokine receptor 2 (CXCR2), with UC. Colocalization analysis further supported these identified links. Differential expression analysis confirmed the dysregulation of the CXCL5/CXCR2 axis in UC patients. Enrichment and immune infiltration analysis indicated that the CXCL5/CXCR2 axis was involved in neutrophil chemotaxis and immune activation in UC. Moreover, CXCL5 expression was found to correlate with neutrophil extracellular trap (NET) formation in UC. Immunohistochemistry further confirmed the dysregulation of the CXCL5/CXCR2 axis in colon tissues of UC patients.

The CXCL5/CXCR2 axis has been implicated to play a significant role within a broader inflammatory network that includes Interleukin (IL)-17, NF-κB, and Tumor Necrosis Factor (TNF) signaling pathways. Additionally, this axis interacts with macrophages and T cells, further contributing to the complexity of inflammatory responses in UC.

There is a significant association between CXCL5/CXCR2 and UC under the MR assumption, which is potentially linked with colonic chemotaxis and activation of neutrophils. These findings highlight the potential of CXCL5/CXCR2 as a therapeutic target for UC. However, future functional studies are needed to validate these findings and explore the exact mechanisms by which CXCL5/CXCR2 influences immune cell crosstalk in UC.

Spirooxindoles have been reported to be effective anticancer drug candidates by displaying promising pre-clinical results. Therefore, to find out a lead spirocyclic oxindole template, a series of spirooxindole derivatives bearing pyrrolizidine (14a-e) and N-methyl pyrrolidine (15a-e) were synthesized using an efficient multicomponent, one-pot, and stereoselective [3+2] cycloaddition reaction and evaluated in vitro against HT29 and HCT116 human colon cancer cell lines.

The pyrrolizidine and N-methyl pyrrolidine spirooxindole derivatives were synthesised in excellent regio- and stereoselectivity using previously optimized reaction conditions. They were evaluated in vitro against cell lines HT29 and HCT116. In silico ADME profiling, molecular docking, and dynamics simulation studies were performed to ascertain the probable mode of action of the lead derivative.

The spirooxindoles were characterized using FTIR, ESI-MS, ¹H and ¹³C NMR, purity was determined by RP-HPLC, and stereochemistry was confirmed by X-ray crystallography. Compound 14a produced the best anti-colon cancer activity with IC50 values of 62.66 and 9.55 µM against HT29 and HCT116 human colon cancer cell lines, respectively. The in silico studies revealed that MDM2 protein inhibition is a probable mode of anti-colon cancer activity, supported by the data obtained in the molecular docking and molecular dynamics study.

The described [3+2] cycloaddition reaction proved to be a highly efficient and catalyst-free reaction. The in vitro cell viability assays and in silico studies revealed that more spirooxindoles can be designed with a varied degree of substitution to target colon cancer.