Idiopathic Pulmonary Fibrosis (IPF) is characterized primarily by progressive lung tissue scarring. This study aimed to explore the molecular mechanism of the HOXA11-AS/miR-148a-3p/SMAD2 axis in IPF. Sixty IPF patients and 70 healthy controls were recruited. mRNA expression was measured by RT-qPCR. An IPF model was established in A549 cells and MRC-5 cells using transforming growth factor-beta 1 (TGF-β1) induction. Cell proliferation, apoptosis, oxidative stress levels, and fibrosis markers were assessed using the cell counting kit-8 (CCK-8), flow cytometry, and enzyme-linked immunosorbent assay (ELISA), respectively. A dual-luciferase reporter assay was conducted to confirm the regulatory interaction. In IPF patients, HOXA11-AS expression was significantly upregulated. Its expression was negatively correlated with lung function parameters as well as with the 6-minute walk test (6MWT) distance. Conversely, it was positively associated with fibrosis-related serum markers, such as hyaluronic acid (HA) and laminin (LN). These findings suggested that HOXA11-AS may serve as a promising diagnostic biomarker for IPF. Moreover, HOXA11-AS promoted the proliferation, suppressed apoptosis, enhanced oxidative stress by reducing superoxide dismutase (SOD) activity and increasing malondialdehyde (MDA) levels, and upregulated the key fibrosis markers expression, including alpha-smooth muscle actin (α-SMA), Collagen I, and Fibronectin. HOXA11-AS directly binds to miR-148a-3p, which in turn targets and regulates SMAD2. Notably, inhibition of miR-148a-3p reversed the suppressive effects of HOXA11-AS knockdown on TGF-β1-induced proliferation, oxidative stress, and fibrotic phenotypes in A549 cells, while restoring SMAD2 expression. Consistent results were observed in MRC-5 fibroblasts: HOXA11-AS knockdown significantly attenuated TGF-β1-induced proliferation, promoted apoptosis, and decreased the expression of α-SMA, Collagen I, and Fibronectin. These effects were partially reversed by miR-148a-3p inhibition, confirming the functional role of the HOXA11-AS/miR-148a-3p/SMAD2 axis in lung fibroblasts. HOXA11-AS promoted the IPF progression by upregulating SMAD2 expression through the sponging of miR-148a-3p.

Viral pneumonia (VP) is a major cause of severe respiratory illness in children. lncRNAs are involved in disease progression by regulating immune regulation. This study investigates the clinical value and regulatory function of exosomal DLEU2 in pediatric VP. Include 247 pediatric VP cases, divided into 120 mild and 127 severe groups, and collected plasma exosomes samples. The abundance of DLEU2, miR-330-5p, and CEBPD were quantified using qRT-PCR. ROC curves and logistic regression were performed to determine the diagnostic and prognostic utility. The correlation was assessed between DLEU2 level and clinical indicators. A VP cell model was established by stimulating BEAS-2B with Poly(I: C). Cell transfection, ELISA, flow cytometry and dual-luciferase reporter assays were conducted to detect cytokines, apoptosis level and target gene regulation. Exosomal DLEU2 was elevated in severe VP and showed good diagnostic performance in distinguishing severe from mild disease. High DLEU2 was independently associated with increased disease severity and correlated with adverse status, including tachypnea, respiratory retractions, reduced SpO2, and elevated inflammatory markers. Poly(I: C) stimulation upregulated DLEU2 and promoted cytokine IL-6 and IL-8 secretion and apoptosis in BEAS-2B. Silencing DLEU2 markedly attenuated these effects. DLEU2 functioned as a ceRNA by sponging miR-330-5p, thereby relieving its inhibitory effect on CEBPD. Restoration of miR-330-5p reversed DLEU2-mediated inflammatory and apoptotic responses. DLEU2 is upregulated in pediatric VP and is associated with disease severity. Functional assays suggest that DLEU2 may aggravate inflammatory injury in bronchial epithelial cells via miR-330-5p.

Acute myocardial infarction (AMI) is a critical and fatal cardiovascular condition. The role of cuproptosis as an emerging mechanism in the pathogenesis of AMI remains to be fully elucidated. Patient data were acquired from the Gene Expression Omnibus database. Differential expression analysis was performed on cuproptosis-related genes (CRGs). LASSO regression was employed to identify key CRGs and develop a diagnostic classification model for AMI. Weighted gene co-expression network analysis was performed to investigate key modules associated with the disease and its molecular classification. Subsequently, machine learning techniques were used to identify critical genes within these modules. The diagnostic efficacy of these genes for AMI was assessed using receiver operating characteristic analysis. The functions of the identified key genes were ultimately validated at the cellular level. Six characteristic CRGs were selected via LASSO regression. The AMI diagnostic classification model based on these CRGs demonstrated superior performance. Patients were classified into two subtypes related to cuproptosis, revealing significant differences in enrichment pathways between these subtypes. SYTL3, SCML1, and PMAIP1 were identified as key genes for AMI, with area under the curves of 0.813, 0.795, and 0.873, respectively. Knocking down PMAIP1 expression reduced intracellular copper levels in hypoxia-induced HUVECs. This study clarifies the role of cuproptosis in AMI and highlights the potential involvement of PMAIP1, providing a theoretical basis for further investigation into cuproptosis in AMI.

14–3-3ζ is becoming an important marker and possible treatment target because many studies link it with cancer recurrence and resistance to chemotherapy. When cancer cells make too much 14–3-3ζ, they tend to survive better, especially under stressful conditions, and they can grow even without attachment, which is usually a sign of more aggressive behavior. On the other hand, when the level of 14–3-3ζ is lowered, tumors grow slower and the cells become more sensitive to different anticancer drugs. In clinical settings high 14–3-3ζ can work as prognostic biomarker, helping to identify patients whose tumors might not respond fit to standard treatments, allowing doctors to think about more personalized therapy options. This protein also interacts with several cancer-related pathways for example working together with HER2, which can push tumors to become more invasive, and harder to treat. Many cancers show amplification of the 14–3-3ζ gene and this usually correlates with worse outcomes for patients. Even though scientists are still figuring out how directly target 14–3-3ζ, a lot of studies suggest that stopping this protein or blocking the signals it activates might make cancer cells respond better in chemo and slow the tumor from getting worse [14]. So overall, going after 14–3-3ζ looks like the pretty promising approach for dealing with therapy resistance, and it could maybe help improve patient outcomes in the future. The 14-3-3 protein family can actually act in two totally opposite ways in cancer, which makes it kinda tricky to study. In some situations, 14-3-3 works like an anti-cancer helper because it stabilizes the p53 protein. When p53 is stable, it can stop the cell cycle and even push damaged cells into apoptosis, so those cells don’t keep dividing. But in other cases, 14-3-3 ends up doing the complete opposite and helps the tumor grow. It mainly does this by turning on the PI3K/AKT pathway, which then causes NF-κB to move into the nucleus. Once NF-κB is in the nucleus, it switches on a bunch of genes that let cancer cells survive longer, divide faster, and even migrate or invade other tissues. So overall, 14-3-3 proteins can either slow cancer down or help it get worse, depending on which signaling pathway is taking over in the cell.

Colorectal cancer (CRC) remains one of the most commonly diagnosed malignancies worldwide and is associated with high recurrence and poor survival outcomes, particularly in its advanced stages. Zi-Shen-Gu-Sui Decoction (ZSGSD), a traditional Chinese medicinal formula, has been clinically applied in CRC treatment and postoperative rehabilitation; however, its pharmacological mechanisms are not yet fully elucidated. This study aimed to explore the anti-CRC efficacy and molecular mechanisms of ZSGSD, with particular attention to the PI3K/AKT/mTOR signaling axis. UPLC-Q-Orbitrap HRMS was utilized to characterize the chemical profile of ZSGSD. Network pharmacology was employed to identify key signaling pathways and bioactive compounds, and molecular docking was conducted to predict potential interactions between major constituents and PI3K/AKT/mTOR proteins. A xenograft mouse model of CRC was established to evaluate tumor suppression. Histological staining was used to assess apoptosis, and the expression of PI3K, AKT, and mTOR was quantified via RT-qPCR, Western blotting, and immunofluorescence assays. Sixty compounds were identified in ZSGSD, with flavonoids comprising approximately 50%. Network analysis suggested potential involvement of the PI3K/AKT/mTOR pathway, and docking results indicated favorable predicted binding between key flavonoids (including Genistein, Apigenin, and Naringenin) and proteins within this pathway. In vivo, ZSGSD treatment suppressed tumor growth in a dose-responsive fashion, with tumor inhibition rates ranging from 30.83% to 75.23% (p < 0.05). Histological evaluations revealed reduced nuclear-to-cytoplasmic ratios and enhanced apoptotic activity, as confirmed by TUNEL staining (p < 0.05). Molecular analyses showed a dose-related reduction in PI3K, AKT, and mTOR expression at the transcript and protein levels, indicating that this pathway may be modulated during ZSGSD treatment (p < 0.05). ZSGSD demonstrates notable tumor-suppressive and pro-apoptotic effects in a CRC xenograft model. These outcomes appear to be associated with modulation of the PI3K/AKT/mTOR signaling pathway, although further functional studies are required to confirm pathway specificity and clarify causal mechanisms.

This study aims to explore the functions and mechanisms underlying the involvement of 14-3-3β in esophageal squamous cell carcinoma (ESCC). Western blot and quantitative real-time PCR (RT-PCR) were utilized to evaluate the levels of 14-3-3β in SHEE, TE-1, and KYSE-150 cells. Following infection with YWHAB-RNAi lentivirus and puromycin screening, the reduction of 14-3-3β expression in KYSE-150 cells was confirmed via western blot and RT-PCR. Subsequently, wound healing, CCK8, and colony-formation assays were performed to assess cell migration and proliferation in KYSE-150 cells with stable low levels of 14-3-3β. Furthermore, western blot analysis was conducted to examine the expression of relevant proteins, elucidating the potential molecular mechanisms underlying 14-3-3β involvement in ESCC progression and metastasis. The expression of 14-3-3β was significantly elevated in KYSE-150 cells compared to SHEE cells. Upon infection in KYSE-150 cells, the expression of 14-3-3β was diminished, resulting in suppressed cell migration and colony formation, although no significant changes were observed in the CCK8 assays. Furthermore, in KYSE-150 cells exhibiting reduced 14-3-3β expression, the activity of phosphorylated AKT (p-AKT) was significantly decreased, while no notable difference was detected in AKT protein levels. Additionally, there were no significant alterations observed in apoptosis-related proteins, including BCL2, BAX, Caspase3, and activated Caspase9. In KYSE-150 cells infected with YWHAB-RNAi lentivirus, the downregulation of 14-3-3β expression resulted in significant reductions in both cell migration and proliferation. Furthermore, the findings demonstrated that 14-3-3β may regulate the biological activities of ESCC cells via modulation of the PI3K/AKT signaling pathway.

Abstract Background MicroRNAs (miRNAs) play a crucial role in tumorigenesis and malignant transformation. Studies indicate that miR-4652-3p is aberrantly expressed in various cancer types. However, its impact and underlying mechanisms in non-small cell lung cancer (NSCLC) have not been investigated. Methods A549 cells were stimulated with IL-1β, TNF-α, and IL-6 (each at 10 ng/ml) to mimic an inflammatory microenvironment. Metabolic status was evaluated by measuring glutamine uptake, α-ketoglutarate (α-KG), and ATP levels. Functional studies employed the glutaminase inhibitor (CB-839), the MYC inhibitor (10058-F4), SLC1A5 small interfering RNA (siSLC1A5-2), a miR-4652-3p mimic, and overexpression plasmids. Molecular interactions were validated using chromatin immunoprecipitation (ChIP), dual-luciferase reporter assays, and RNA pull-down experiment. CCK-8 and Transwell assays were used for the assessment of cell malignant phenotypes. The functional significance of miR-4652-3p was further verified in a xenograft mouse model. Results miR-4652-3p was downregulated in NSCLC, while MYC and SLC1A5 were upregulated. Inflammatory stimulation enhanced A549 cell proliferation, glutamine uptake, and α-KG/ATP production; these effects were attenuated by CB-839. ChIP and dual-luciferase assays demonstrated that MYC binds the SLC1A5 promoter and activates its transcription. Inhibiting MYC or knocking down SLC1A5 significantly reduced glutamine uptake. Mechanistic analysis revealed that miR-4652-3p directly targets both MYC and SLC1A5 mRNA. miR-4652-3p suppressed glutamine metabolism in NSCLC cells by negatively regulating the MYC/SLC1A5 axis, consequently inhibiting cell growth and tumor progression in a xenograft mouse model, an effect reversed by MYC or SLC1A5 overexpression. Conclusions miR-4652-3p blocked the inflammatory microenvironment-induced glutamine metabolic reprogramming by directly suppressing the MYC/SLC1A5 axis, thereby inhibiting NSCLC progression. The miR-4652-3p/MYC/SLC1A5 pathway represents a key regulatory mechanism for metabolic adaptation in NSCLC.

BackgroundAtherosclerosis (AS) is a leading cause of cardiovascular-related death worldwide. The role and regulatory mechanism of GAS6-AS2 in AS remain unclear.AimTo investigate the diagnostic/prognostic value of GAS6-AS2 in AS and clarify its molecular mechanism.Methods107 AS patients and 105 healthy controls were included. The levels of GAS6-AS2, miR-138-5p, and mRNA were measured using RT-qPCR. ROC curve, K-M survival analysis, and Cox regression were performed to evaluate the diagnostic and prognostic value of GAS6-AS2. Bioinformatics prediction and dual-luciferase reporter assay were performed to verify the regulatory axis. Ox-LDL-induced VSMCs were used to construct an AS cell model. The biological functions were assessed using CCK-8, SA-β-Gal, and ELISA.ResultsGAS6-AS2 expression was significantly increased in AS patients and in VSMCs treated with ox-LDL, and it showed high diagnostic accuracy and risk prediction for patients with AS. Knockdown of GAS6-AS2 reduced SA-β-Gal-positive cells, downregulated the expression of senescence-related genes and proteins (p16, p21, p53), and decreased the levels of inflammatory factors (IL-6, IL-1β) in ox-LDL-induced VSMCs. Mechanistically, GAS6-AS2 directly bound to miR-138-5p and inhibited its expression, while miR-138-5p targeted AKT1 to suppress its expression. Rescue experiments confirmed that the GAS6-AS2/miR-138-5p/AKT1 axis mediated ox-LDL-induced VSMC senescence and inflammation.ConclusionsGAS6-AS2 is a potential diagnostic and prognostic biomarker for AS. It regulates ox-LDL-induced VSMC senescence and inflammatory response through the sponging of miR-138-5p to upregulate AKT1, providing a novel molecular target for AS treatment.Supplementary InformationThe online version contains supplementary material available at 10.1186/s41065-026-00650-5.