PurposeTo investigate the genomic resistance profile of carbapenem-resistant Acinetobacter baumannii (CRAB) isolates from ICU environments, with a focus on characterizing a representative CRAB strain I2 to elucidate its genomic determinants of resistance and assess their implications for infection control.MethodsBetween 2012 and 2015, a total of 24 Acinetobacter baumannii strains were isolated from high-touch surfaces ICUs of four hospitals. Antimicrobial susceptibility testing against 15 antibiotics was performed for all isolates using the VITEK® 2 system. One representative strain was selected for whole-genome sequencing. Resistance genes, virulence factors, and mobile genetic elements were systematically analyzed using bioinformatics tools and databases. In addition, the biofilm formation capacity of this strain was quantitatively assessed by crystal violet staining.ResultsResistance rates to β-lactams ranged from 58.33% to 66.67%, while 95.83% of isolates remained susceptible to polymyxin. The representative CRAB strain I2 (sequence type 191) harbored three carbapenemase genes and 13 ade efflux pump genes, with 40 resistance genes identified (68.75% efflux-mediated). Genomic island GI16 (carrying transposase ISAba1) suggested horizontal gene transfer driving resistance dissemination. A total of 99 virulence genes and disinfectant resistance genes were detected. Biofilm formation capacity was moderate. Genomic analysis of strain I2 revealed a comprehensive resistance profile and potential mechanisms underlying environmental persistence and transmission.ConclusionThe ICU environment constitutes an important reservoir for CRAB. The strain I2 harbored key resistance determinants, including efflux pump, and mobile genetic elements, which correlated with its carbapenem-resistant phenotype. Additionally, this strain harbors biofilm-associated genes and disinfectant efflux pump genes, and exhibits moderate biofilm-forming capacity, indicating strong environmental adaptability. The genomic characteristics of strain I2 provide a molecular basis for implementing targeted CRAB infection control strategies in high-risk healthcare settings.

Recurrent vulvovaginal candidiasis (RVVC) is a common, refractory fungal infection affectingwomen, primarily caused by Candida albicans. The interplay among fungal virulence factors, biofilm formation, and antifungal resistance is crucial in the pathogenesis of RVVC. This study compared 50 Candida albicans isolates from RVVC patients and 50 from asymptomatic vaginal colonizers. Antifungal susceptibility testing was performed using the broth microdilution method. Biofilm formation was assessed via crystal violet staining, and the expression levels of virulence factor hydrolases (SAP, PL, Lip) and cell wall protein genes (ALS1, ALS3, HWP1) were analyzed using phenotypic assays and quantitative real-time PCR (qRT-PCR). Pearson correlation analysis was used to evaluate the relationships among these parameters and antifungal resistance. RVVC isolates exhibited significantly higher MICs for fluconazole, voriconazole, and itraconazole. Biofilm formation ability and the expression levels of SAP, PL, Lip, ALS1, ALS3, and HWP1 were also significantly higher in RVVC isolates. A moderate correlation was observed between antifungal drug MIC values and biofilm OD, while a weak correlation existed between MIC values and ALS/HWP1 gene expression. Notably, hydrolase expression showed no significant correlation with resistance. Candida albicans from RVVC patients demonstrated enhanced biofilm formation, virulence factor expression, and antifungal resistance. Biofilm-mediated drug tolerance may be a key mechanism underlying the refractoriness of RVVC. Targeting biofilm formation and virulence factor genes may offer novel strategies for managing RVVC.

Background: Salinity is a critical threat to agricultural productivity, reducing crop yield and soil fertility in arid and semi-arid regions. Conventional remediation methods such as leaching and chemical desalination are expensive, unsustainable, and environmentally harmful. Therefore, the development of biological approaches using halotolerant bacteria provides a sustainable alternative for reducing salinity and enhancing crop growth under salt stress. Objective: This study aimed to isolate and characterize halotolerant bacterial strains from coco-coir and evaluate their potential to form biofilms for desalination and plant growth promotion under saline conditions. Methods: Coco-coir samples were aseptically collected from Lahore Garrison University and cultured on nutrient agar supplemented with 1M, 2M, and 3M NaCl to isolate halotolerant bacteria. Morphological and biochemical characterization, including Gram staining, catalase, oxidase, citrate, indole, and Voges–Proskauer tests, were conducted for identification. Six isolates (AK-1 to AK-6) were screened for biofilm formation using the crystal violet assay. Isolate AK-6, showing the highest tolerance (growth up to 2M NaCl) and strongest biofilm development, was selected for a pilot-scale desalination experiment using 1M artificial seawater with coco-coir and sand as substrata. Zea mays seeds inoculated with five isolates (AK-1, AK-3, AK-4, AK-5, and AK-6) were evaluated for germination and growth parameters under both saline and non-saline conditions. Results: All isolates formed distinct colonies and biofilms. AK-6 demonstrated strong growth at 2M NaCl and weak growth at 3M NaCl. The pilot setup showed a 46% reduction in Na⁺ concentration over 10 days. Inoculated Zea mays plants exhibited 85% germination under 1M saline conditions compared to 0% in non-inoculated controls. Mean shoot and root lengths increased by 73% and 81%, respectively, in inoculated plants, while chlorophyll a and b levels improved significantly (p < 0.05) compared to controls. Conclusion: Halotolerant bacterial isolates, particularly AK-6, demonstrated effective biofilm-mediated desalination and promoted Zea mays growth under salinity stress. The integration of such bacterial biofilms offers a cost-effective and eco-friendly solution for managing saline soils and water in agriculture.

Enhanced structural and optical properties of MgFe2O4 spinel ferrite by gamma irradiation for crystal violet and bromophenol blue dyes removal from wastewater

Synthesis and Characterization of CuFe2O4–ZnO Nanocomposite via Ultrasonication-Assisted Microwave Combustion for Photocatalytic Degradation of Crystal Violet Dye

Abstract Microplastics (MPs) are recognized as emerging pollutants in aquatic environments, where they are rapidly colonized by microbial communities that form biofilms. These biofilms can alter the environmental behaviour, transport characteristics, and ecological impact of MPs. Although many studies have simulated biofilm formation under laboratory conditions, fewer have examined natural biofilm development on MPs in freshwater systems. This study investigates biofilm formation on polystyrene (PS) MPs (shape: fragments) in different flow conditions of a natural riverine setting. The goal is to develop a protocol for producing environmentally relevant biofilm-coated MPs. PS MPs of two size classes, namely 20–75 $$\upmu$$ m and 600–1000 $$\upmu$$ m, were incubated for four weeks in the Rhine River using a perforated box (low flow environment) and a tube setup (high flow environment). Immersion microscopic observations revealed widespread microbial colonization across all MPs, with higher flow conditions supporting visibly more surface coverage and diverse biofilms. Scanning electron microscopy revealed the presence of various microorganisms-including diatoms, bacteria, ciliates, and choanoflagellates-in the high-flow tube setup, whereas they were largely absent in the low-flow box setup. Spectrophotometric analysis using crystal violet staining confirmed significantly higher biofilm biomass (higher absorbance values) in the tube incubation setup (0.2374 ± 0.0865) compared to the box setup (0.0764 ± 0.0225). The results demonstrated that flow velocity plays a critical role in shaping biofilm density and microbial composition. Higher flow conditions likely promoted greater nutrient exchange and surface contact, facilitating enhanced colonization. These findings underscore the importance of mimicking realistic hydrodynamic conditions when preparing biofilm-coated MPs for environmental studies. The methodology developed in this study is a step towards a standardized approach to generating environmentally relevant MPs, which can improve the accuracy of future research on MP behaviour, transport, and ecological interactions.

Surface-enhanced Raman scattering (SERS) has emerged as a powerful analytical tool for ultrasensitive trace molecule detection. Herein, we report the fabrication of uniform nanofilm substrates composed of Au–Ag alloy hollow nanoparticles (HNPs) with tunable sizes, demonstrating exceptional performance as SERS platforms. Using crystal violet (CV) as a Raman probe, the substrates exhibit densely and uniformly distributed “hot spots” across the nanofilm surface, enabling a detection limit as low as 10[Formula: see text] M for CV with excellent signal reproducibility (relative standard deviation, RSD [Formula: see text] 6.61%). Notably, the substrates achieve quantitative detection of thiram pesticides at ultratrace concentrations (0.01 ppb). Finite-difference time-domain (FDTD) simulations elucidate that the observed SERS enhancement originates from the synergistic contribution of two factors: (1) the localized surface plasmon resonance (LSPR) effects mediated by the Au–Ag HNPs and (2) the formation of abundant, spatially homogeneous “hot spots” within the nanofilm architecture. This study highlights the potential of size-engineered Au–Ag HNP nanofilms as robust, and high-performance SERS substrates for environmental and analytical applications.

Mycoremediation emerges as an alternative strategy for decolourisation of synthetic dyes and is valued for its cost-effectiveness and environmentally friendly attributes. Five fungal strains, Aspergillus sp.1, Lasiodiplodia crassispora, L. pseudotheobromae, Neopestalotiopsis saprophytica, and Trichoderma sp.1, isolated from freshwater environments in Sri Lanka, were subjected to decolourisation of 100 mg L-1 of Congo Red (CR), Crystal Violet (CV), Malachite Green (MG), and Safranin dyes, frequently discharged into the environment from laboratories and industries. Screening of the decolourisation ability of isolated fungal strains was conducted in both solid and liquid media containing Potato Dextrose Agar (PDA) and Potato Dextrose Broth (PDB) for ten days incubation period and 14–28 days, respectively, at 30 °C. The liquid media screening processes showed that L. pseudotheobromae exhibited the highest decolourisation percentage for CV (95.23% ± 0.82) and MG (93.12% ± 0.36). L. crassispora demonstrated the highest decolourisation abilities for CR (91.45% ± 0.20) and Safranin. All fungal strains successfully achieved over 60% decolourisation of CV, CR, and MG. However, Safranin showed the lowest decolourisation by all isolated strains, except for L. crassispora (70.46% ± 1.18). Considering the overall results in both solid and liquid media (exceeding 70%), L. crassispora exhibited the highest decolourisation ability among all selected dyes. Besides, the results in liquid media were reconfirmed by the screening process on solid media. The results of the present study showed that mycoremediation for synthetic dye decolourisation should be expanded to outdoor settings. Leveraging this insight, a prototype was developed for real-world application, creating a microencapsulation system for mycoremediation. This innovative system offers a sustainable alternative to traditional physicochemical treatments for wastewater management, specifically on laboratory discharges.

‏A direct comparison was made between raw banana peel waste (RBPW) and acid-treated banana peel waste (ABPW), under identical conditions, for adsorption of crystal violet (CV). Sorption kinetics, isotherms, and thermodynamics were considered to reveal the underlying mechanisms. The effects of contact time, pH, initial CV concentration, temperature, and adsorbent dosage were evaluated. The sorption process obeyed a pseudo-second-order kinetic model, while the Langmuir isotherm model best explained the equilibrium data with maximum adsorption capacities. The Dubinin–Radushkevich model supported the potential of ion-exchange mechanisms for the acidified sample. Adsorption was spontaneous and endothermic, as revealed by negative Gibbs free energy, positive enthalpy (+16.4 kJ/mol for RBPW and +53.5 kJ/mol for ABPW) and positive entropy (RBPW = 6.79 J/mol·K and ABPW = 14.65 J/mol·K) values. The lower ΔH for the raw peel is more consistent with physisorption, while higher ΔH of the acid-treated peel suggests stronger interactions consistent with chemisorption/ion-exchange. The FT-IR analysis confirmed that functional groups such as –OH, –COOH, C=O, C-O, and possibly aromatic moieties on banana peel waste are involved in the sorption of CV. The enhanced performance of ABPW is attributed to acid-induced surface modifications that increased porosity, making the functional groups available for sorption process.

Despite the established clinical efficacy of oxaliplatin in colorectal cancer (CRC), resistance to this platinum-based agent continues to pose a significant therapeutic challenge. Increased exportin 1 (XPO1) expression in CRC has been linked to chemoresistance, while KPT-330, a selective XPO1 inhibitor, has exhibited potential in enhancing platinum drug effectiveness in other cancer types. The present study explored the synergistic effects of KPT-330 and oxaliplatin in oxaliplatin-resistant CRC models. Oxaliplatin-resistant cell lines (HCT116/L-OHP and HCT8/L-OHP) were developed, exhibiting elevated XPO1 expression as demonstrated by western blotting. A range of in vitro assays (Cell Counting Kit-8 assays, ethynyldeoxyuridine assays, crystal violet staining, transmission electron microscopy and flow cytometry) and an in vivo subcutaneous xenograft model in nude mice were used to evaluate the combination therapy. Co-treatment with KPT-330 and oxaliplatin induced G2/M phase arrest and mitochondrial dysfunction, thereby triggering apoptosis and ferroptosis. Mechanistically, the combination therapy of KPT-330 and oxaliplatin promoted the nuclear retention of p53, which in turn upregulated p21 and downregulated solute carrier family 7 member 11. In vivo, the combination therapy significantly enhanced tumor sensitivity to oxaliplatin. These results suggested that KPT-330 restored oxaliplatin sensitivity in resistant CRC by facilitating p53 nuclear retention, presenting a promising approach to overcome chemoresistance through dual modulation of cell cycle arrest and ferroptosis pathways.

Platelet concentrates (PCs) are stored at 20–24˚C in a biologically favorable environment that may support bacterial growth. Staphylococcus epidermidis, a typical contaminant, can form biofilms in PCs, complicating detection and increasing the risk of transfusion-transmitted bacterial infections. The material composition and surface texture of PC storage bags may influence biofilm formation. The impact of different PC storage bag materials on S. epidermidis biofilm formation was evaluated using the ISO 4768:2023(E) crystal violet (CV) assay. Four surface conditions were tested: polyvinyl chloride (PVC) plasticized with n-butyryl-tri(n-hexyl)-citrate (BTHC) – both smooth and rough sides, PVC plasticized with tri-(2-ethylhexyl)-trimellitate (TEHTM) and ethylene-vinyl acetate (EVA). Coupons and bags made from each material were used in the experiments. Biofilm-positive S. epidermidis was cultured in tryptic soy broth (TSB), PCs and plasma and added on plastic coupons under static conditions or directly in the bags with agitation. Bacterial enumeration and CV assay were performed on days 2, 5, and 7. In TSB, EVA coupons significantly formed more biofilm than the smooth side of PVC-BTHC or TEHTM over seven days. In PCs, more biofilm formed on the rough side of PVC-BTHC coupons than the smooth side, with no other differences between plastics, suggesting similar biofilm amount across PC bag materials in the presence of platelets. No biofilm was detected on coupons in plasma. Under continuous agitation and reduced oxygen levels, only the rough side of PVC-BTHC showed significant biofilm formation in TSB in PC storage bags over seven days. These findings highlight the need for standardized biofilm testing and suggest that some plastics are more conducive to biofilm formation under static conditions. However, during blood bank storage (i.e., continuous agitation and reduced oxygen levels), biofilm formation is limited, regardless of the platelet bag material, thereby reducing the risk of undetected bacterial contamination.

The challenges related to biofilm-associated infections and diseases have prompted scientists to identify the factors responsible for the formation of biofilms and to develop strategies aimed at decreasing biofilm-formation capacity. The present study aimed to evaluate the effect of randomly methylated cyclodextrin derivatives, at concentrations ranging from 32 nM to 12.5 mM, on the capacity of Candida albicans to form biofilms at 37 °C over a period of 24 h. This study provides novel insights into how randomly methylated α-cyclodextrin (RAMEA), randomly methylated β-cyclodextrin (RAMEB), and randomly methylated γ-cyclodextrin (RAMEG) can modulate C. albicans biofilm formation. Using the crystal violet assay and the XTT reduction assay, we consistently demonstrated that RAMEA and RAMEG have a clear, concentration-dependent inhibitory effect on both the total biofilm biomass and the metabolic activity of the cells associated with these biofilms. RAMEB exhibited a biphasic effect: low to moderate concentrations significantly reduced biofilm formation, while higher doses unexpectedly resulted in increased biofilm formation. Microscopic analysis revealed that elevated cyclodextrin concentrations induced hyphal formation. Optical density measurements and a membrane permeability assay indicated that none of the cyclodextrins had a notable cytotoxic effect or damaged the cell membrane. Moreover, elevated intracellular ROS levels were detected, suggesting a potential stress-inducing effect. These findings enhance our understanding of the complex interactions between cyclodextrin derivatives and fungal cells, underscoring their potential as biofilm-modulating agents.

Traditional Chinese medicine Pudilan synergizes with smc gene to impair cell growth and exopolysaccharides synthesis of Streptococcus mutans.

Carbapenem-resistant Pseudomonas aeruginosa(CRPA) poses a serious threat in healthcare settings due to its multidrug resistance and high mortality. Although ceftazidime/avibactam (CZA) demonstrates potent activity against CRPA, resistance has emerged.ObjectiveThis study investigates the epidemiology and molecular mechanisms of CZA resistance in CRPA isolates from Ningbo, China.MethodsA total of 279 non-duplicate clinical CRPA isolates (2022–2024) were classified as CZA-resistant (CZA-R, n = 68) or CZA-susceptible (CZA-S, n = 211). Carbapenemase genes were detected by PCR, clonality via MLST, biofilm formation by crystal violet assay, and efflux pump expression (mexA, mexC, mexE, mexY) via qRT-PCR. WGS was performed on selected isolates.ResultsThe CZA resistance rate was 24.37%. Risk factors included recent trauma, prior antibiotic exposure, central venous catheterization, and drainage tube placement (all p < 0.05). The CZA-R group showed higher recurrence (13.2% vs. 4.3%, p = 0.029) and lower clinical improvement (67.6% vs. 77.3%, p = 0.029). blaNDM prevalence was higher in CZA-R (7.4% vs. 0.5%, p = 0.003), and ST1076 was the predominant clone (29.3%), with higher representation in CZA-R (40.0%). Horizontal gene transfer mediated blaNDM spread. CZA-R isolates exhibited enhanced biofilm formation (p < 0.001) and mexA upregulation (2.04-fold, p = 0.007).ConclusionOur findings indicate a high prevalence of CZA resistance among CRPA isolates in Ningbo, driven by multiple mechanisms including blaNDM carriage, enhanced biofilm formation, and overexpression of efflux pumps. The dissemination of the high-risk clone ST1076 underscores the need for strengthened infection control measures to curb its spread. These findings provide important insights for optimizing infection control and treatment strategies against CRPA infections in this region.

Bacterial populations untiringly shift between sessile and motile states in order to adapt and survive during exposition to various stress conditions. From a practical viewpoint, elucidating factors that trigger biofilm formation in non-pathogenic bacteria has a big perspective as soon as representatives of normal colon microflora are known to positively contribute to an overall state of human health. Effects of low molecular thiols on bacterial motility and/or biofilm formation have been poorly studied. The aim of this work was to elucidate contribution of components of thiol metabolism in biofilm formation in E. coli using a number of knock-out mutants in the genes, encoding components of thiol metabolism, stages of biofilm formation, global regulators of responses to stationary-phase stress (RpoS), shift to sessile mode (YdeH), anaerobic shift (ArcB/ArcA), SOS-response (RecA), stringent response (RelA). Motility tests were performed on 0.2% agar plates with LB, after 6 h of incubation at 30⁰C diameters of the swimming zones were measured. Under our conditions, most striking inhibiting effects on swimming ability were found in case of cydD, gor trxB, arcB and recA mutants, when zone diameter decreased by about 7 times compared to the untreated control (28.5 ± 2.6 mm). While mutations in ggt, gsiA ggt, relA and rpoS resulted in about 2 times diminishment of the swimming zones. Deficiency in synthesis of one of the matrix components (pgaA) resulted in just 1.5 times inhibition of the zone. Collectively, it revealed involvement of thiol metabolism as well as participation of the studied global regulators in the processes of planktonic/sessile state transitions. In some cases, the observed down-regulation of the swimming ability coincided with the ability to form biofilms in the crystal violet staining tests. Further research is being carried out.

Quinine (QN), a widely used alkaloid in food and pharmaceuticals, can pose health risks at elevated levels, necessitating the development of sensitive and rapid detection methods. Here, we report a label-free, ratiometric fluorescent aptasensor for the sensitive and selective detection of QN. The sensing platform integrates a quinine-specific DNA aptamer (AptQN) with crystal violet (CV), a triphenylmethane (TPM) dye exhibiting near-infrared (NIR) emission, and fluorescein isothiocyanate (FITC) as an internal reference. Upon QN binding, CV is competitively displaced from the aptamer complex, resulting in fluorescence quenching, while FITC emission remains stable, facilitating self-calibrated quantification. The aptasensor exhibits high sensitivity, rapid response within one minute and robust analytical performance in complex samples, such as lake water and soft drinks. This cost-effective and efficient sensing strategy provides a reliable approach for QN monitoring in food safety and environmental applications.

Gram-positive enterococci are opportunistic and resistant to many antibiotics. This search investigated the prevalence of macrolide antibiotic resistance genes in local enterococcal isolates and its correlation with biofilm formation. We collected 112 clinical samples from the Medical City Hospital, dentists' clinics, and labs in Baghdad from October 2022 to March 2023, which included root canal samples from 50 patients and urine samples from 62 patients with urinary tract infections. The samples were cultured on Pfizer-specific Enterococcus media. Twenty-one isolates were identified as Enterococcus spp. by biochemical tests and confirmed using the VITEK 2 system. After that, the crystal violet staining method was used to assess enterococci isolates' ability to form biofilms in a polystyrene microtiter, and then molecular detection was done to detect the ermB gene. The results revealed that the percentage of enterococcal isolates positive for the ermB gene was 87.5% in root canal samples. In urine samples, the percentage of enterococcal isolates with the same gene was 84.6%. All isolates succeeded in forming biofilm; for urine isolates, 77% and 23% of isolates formed moderate and strong biofilm, respectively. While for root canal isolates, 12.5%, 75%, and 12.5% of isolates formed weak, moderate, and strong biofilm, respectively. We conclude in this study that the ermB gene was detected in enterococcal isolates from the tooth root canal and urine samples, with a higher prevalence percentage in urine sample isolates than tooth root canal isolates. Finally, the findings demonstrated that there is no connection between this gene's existence and the tested isolates' ability to create biofilms.

Coin-metal@semiconductor nanohybrids have attracted extensive attention in surface-enhanced Raman scattering (SERS) detection, aiming to achieve thesynergistic effectofelectromagnetic enhancement mechanism (EM) and chemical enhancement mechanism (CM). In this work, positively charged thin-layered molybdenum disulfide nanosheets (MoS2NSs) and negatively charged gold nanoparticles (AuNPs) were synthesized, which were further fabricated into MoS2NSs@AuNPs via electrostatic interactions and Au-S bonding. The approach is quite rapid, cost-effective, and environmentally friendly, and the resulting MoS2NSs@AuNPs exhibited an outstanding SERS enhancement factor of up to 2.33 × 108 for crystal violet due to the synergistic effect of EM and CM. Furthermore, the fabricated SERS substrate demonstrated long-term stability, maintaining about 80% of SERS activity even after storage for two months, and excellent reproducibility, with a relative standard deviation of 5.20%. Remarkably, this SERS-active nanohybrid enabled rapid and highly sensitive detection of thiram residues in apple juice, achieving satisfactory recoveries of 82.6-110.7% and an impressively low limit of quantification of 50µg/L. This work not only presents a novel strategy for designing high-performance SERS nanomaterials but also highlights their promising applications in food safety monitoring and environmental analysis.

Dyes play an essential role in water contamination; however, more effort must be made to reduce this issue. This study utilised Lagurus ovatus as an adsorbent, which can be considered an inexpensive weed, to remove the crystal violet from aqueous media. Various factors were examined, and it was found that the highest dye removal efficiency was obtained at pH between 5─11, the adsorbent weight of 2 g/L, the contact time of 40 min, and at the temperature of 50 °C. At the same time, it decreases with increasing dye concentration to reach its lowest value of about 82% at 80 mg/L. The thermodynamic study denotes the endothermic behaviour of the removal system. Also, the kinetic investigation elucidates that the best fit of the removal data can be offered by applying the pseudo-second order kinetic equation. In addition, the study of the isotherms reveals that the removal system follows precisely the Langmuir isotherm. The proposed adsorption mechanism is the electrostatic attraction between the cationic dye and the biosorbent surface. These results suggest that Lagurus ovatus, a new and alternative natural adsorbent, can be utilised efficiently to eliminate crystal violet from aqueous media.

ObjectiveTo investigate the regulatory role of MACF1 and its upstream transcriptional control in focal adhesion remodeling and tumor progression in lung adenocarcinoma (LUAD).MethodsWe employed in vitro loss- and gain-of-function assays using shRNA-mediated knockdown and ectopic overexpression of MACF1 and NR2F1 in LUAD cell lines (H1299 and Calu-3). Cell proliferation, adhesion, and migration were assessed by CCK-8, EdU, crystal violet, and Transwell assays. In vivo tumor growth and metastasis were evaluated using subcutaneous and tail vein xenograft models in nude mice. RNA-seq and GSEA were performed to identify MACF1-regulated pathways, followed by nuclear-cytoplasmic fractionation, dual-luciferase reporter assays, and immunofluorescence to assess WNT/β-catenin activity. ChIP-qPCR and ChIP-seq data from ENCODE were used to validate NR2F1 binding to the MACF1 promoter.ResultsMACF1 knockdown significantly suppressed LUAD cell proliferation, DNA replication, adhesion, and migration, and reduced tumor burden and lung metastases in vivo. Mechanistically, MACF1 activated WNT/β-catenin signaling by promoting CTNNB1 nuclear translocation, which upregulated focal adhesion genes (Paxillin, FAK, ITGB1). CTNNB1 agonist TWS119 restored focal adhesion in MACF1-deficient cells. Bioinformatic prediction and ChIP validation identified NR2F1 as a transcription factor directly targeting the MACF1 promoter. NR2F1 deficiency reduced MACF1 expression and phenocopied its functional loss, while MACF1 overexpression rescued the impaired phenotype.ConclusionOur study uncovers a previously unrecognized NR2F1–MACF1–WNT axis that drives focal adhesion formation and LUAD progression. Targeting this regulatory circuit may offer new avenues for anti-metastatic therapy in lung adenocarcinoma.HighlightsNR2F1 is identified as a direct upstream transcription factor that activates MACF1 expression in LUAD.MACF1 promotes LUAD cell proliferation, adhesion, and migration by enhancing focal adhesion assembly.MACF1 activates the WNT/CTNNB1 signaling cascade, facilitating CTNNB1 nuclear translocation and downstream target expression.Loss of MACF1 impairs focal adhesion formation and metastatic potential both in vitro and in xenograft and tail vein models.The NR2F1–MACF1–WNT axis represents a novel regulatory circuit driving LUAD metastasis and offers potential therapeutic targets.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40001-025-03332-6.