Cellular polarity plays a fundamental role in tissue organization and homeostasis, and its disruption is closely linked to tumorigenesis. Crumbs3 (CRB3), a conserved polarity protein, is essential for epithelial morphogenesis, tight-junction formation, and barrier function. This review summarizes current knowledge regarding CRB3 expression in normal and malignant human tissues and its dual roles in cancer progression. Systematic immunohistochemical analyses revealed strong CRB3 expression in non-neoplastic glandular epithelia of the gastrointestinal, hepato-pancreato-biliary, renal, and respiratory tracts, as well as in fetal tissues, suggesting its importance in organ development and maintenance. In neoplastic tissues-represented by colorectal adenocarcinoma and oral squamous cell carcinoma-CRB3 expression is preserved or even enhanced compared with normal tissues, which promotes tumor cell migration, triggering invasion/metastasis as well as cellular proliferation through signaling pathways involving FGFR and RhoA activation. Conversely, previous studies reported that CRB3 functions as a tumor suppressor, based on findings that CRB3 expression induces loss of epithelial-mesenchymal transition, whereas loss of CRB3 expression attenuates the integrity of tight junctions, resulting in significantly poorer prognosis in certain cancers. Current data thus suggest that the biological role of CRB3 in tumors is complex. Whether CRB3 acts as a tumor accelerator or suppressor may depend on the individual-specific, unique characteristics of tumor cells. Understanding these dual functions may contribute to the development of novel polarity-targeted therapeutic strategies for cancers of differing origin.

Tumour seeding after transbronchial biopsy: Mechanisms, clinical implications, and prevention strategies-A narrative review.

Synthesis of a fluorescent probe for the selective detection of cysteine and its application in cell imaging.

Neoadjuvant immunotherapy for resectable esophageal cancer: Research progress and clinical perspectives.

Microfluidic mixing is a widely used technology for producing lipid nanoparticles (LNPs) that encapsulate messenger RNA (mRNA). One major challenge in advancing LNP-based RNA therapeutics consists of developing formulations that can be manufactured consistently across various scales of drug development. Even though microfluidics allows LNP preparation with precisely defined properties, manufacturing at a large scale is very limited and often achieved by parallelizing microfluidic devices (numbering up). Here, we demonstrate the use of a fluidic oscillator as a novel micromixing principle (FDmix) for the preparation of mRNA-LNPs. In this study, we investigated the feasibility of upscaling the production of eGFP mRNA-LNPs by increasing the size of the fluidic oscillator geometry and by adjusting flow parameters. We evaluated the comparability of mRNA-LNP batches, which were either produced using FDmiX S for low- (< 0.8L/h) or FDmiX M for high-output production (< 4.8L/h), via physicochemical properties (Z-Average, PDI, Cryo-TEM, encapsulation efficiency, lipid composition analysis via HPLC). Additionally, mRNA-LNPs were formulated using a T-junction under the same flow parameters to assess potential differences. FDmiX samples were tested for short- and long-term storage stability under different conditions (8°C, -80°C). Small LNPs (< 80nm) with narrow distribution (PDI<0.1) and high encapsulation efficiency (> 96%) were obtained with FDmiX. The biological functionality was shown in vitro by transfection experiments of colorectal adenocarcinoma cells DLD-1 using fluorescence imaging, flow cytometry, and Western Blot. Our results indicate the high potential of the FDmix technology for the production of mRNA nanoparticles. Already at a small scale, high volumes can be processed.

Chronic polystyrene microplastics exposure promotes lung adenocarcinoma metastasis through EREG-regulated phosphorylation-dependent NF-κB activation

Lung cancer is a leading cause of mortality worldwide, with current treatments facing challenges such as adverse effects, limited targeting and resistance. Quercetin, a naturally occurring and abundantly available flavonoid in the global flora, exhibits promising anti-inflammatory and anti-cancer properties. However, its clinical application is limited by poor bioavailability and stability. Nanotechnology offers a solution to enhance the delivery and efficacy of quercetin. In this study, an optimised nanoemulsion of quercetin was developed and its anti-cancer effects against non-small cell lung cancer (NSCLC) were evaluated in the human adenocarcinoma cell line (A549), comparing its effects with the Quercetin pure drug (QPD) at varying concentrations. QNE in comparison to QPD was found to significantly inhibit key cancer hallmarks like cell proliferation, migration and colony formation in the functional assays and downregulated the expression of numerous oncogenic proteins and genes that are associated with angiogenesis, metastasis and uncontrolled cell growth, ultimately facilitating cancer progression. The anticancer potential of Quercetin was further examined through computational approaches involving network analysis and molecular docking that further supported its potential as an anti-cancer agent, correlating in vitro and in silico data. The enhanced anti-cancer activity of QNE compared to QPD suggests that nano-emulsion based delivery of quercetin is attributing to its effective therapeutic efficacy, highlighting its potential against lung cancer. However, expedited studies, particularly in vivo animal models, are essential to fully assess its therapeutic potential and safety profile.

Revealing physical properties of gastric adenocarcinoma cells with two distinct morphologies linking to preferential cellular migration and proliferation.

Osimertinib is the standard first-line therapy for EGFR-mutant lung adenocarcinoma; however, the inevitable development of acquired resistance leads to disease progression and treatment failure. While established resistance mechanisms primarily involve genetic alterations, stress-adaptive pathways, particularly stress granule-mediated therapeutic tolerance, remain poorly understood. This study aims to elucidate the transcriptional and post-translational mechanisms governing stress granule-mediated survival and their contribution to Osimertinib resistance in lung adenocarcinoma. We identify NF-κB repressing factor (NKRF) as a critical suppressor of Osimertinib resistance, whose expression is markedly reduced in resistant lung adenocarcinoma cells. Restoration of NKRF significantly sensitized resistant cells to Osimertinib in vitro and inhibited tumor growth in xenograft models. Mechanistically, NKRF directly repressed transcription of the ribonucleoprotein component Small nuclear ribonucleoprotein D2 (SNRPD2), thereby constraining stress granule formation and attenuating drug tolerance. We further demonstrate that the E3 ubiquitin ligase TRIM26 interacts with NKRF and promotes its K48-linked ubiquitination at Lys411, leading to proteasomal degradation. This process sustains SNRPD2 expression and enhances stress granule assembly. Genetic depletion of TRIM26 restored NKRF stability, suppressed stress granule formation, and re-sensitized resistant tumors to Osimertinib, effects that were abrogated by concomitant NKRF silencing. Collectively, this study defines a previously unrecognized TRIM26/NKRF/SNRPD2 regulatory axis that integrates ubiquitin-mediated proteostasis with transcriptional control of stress granule dynamics. This work provides mechanistic insight into stress-adaptive Osimertinib resistance and identifies potential therapeutic targets for overcoming resistance in EGFR-mutant lung adenocarcinoma.

The predictive value and clinical application of measuring circulating endothelial progenitor cells in lung adenocarcinoma.

ABSTRACT Chimeric antigen receptor (CAR)‐T cells generated using ephrin‐B2, the natural ligand for ephrin type‐B receptor 4 (EphB4), have demonstrated antitumor activity; however, their efficacy remains unvalidated. Therefore, we evaluated the dual‐targeting antitumor ability of these cells and confirmed their efficacy against lung adenocarcinoma. First, we evaluated EphB4 and EphA2 expression in samples from 74 patients with lung adenocarcinoma using immunohistochemistry. Next, EphB4 CAR‐T cells were generated via piggyBac ‐mediated gene transfer, and their phenotype was evaluated. Subsequently, we conducted an antigen stimulation assay to assess the bispecificity of the EphB4 CAR‐T cells. Finally, the antitumor effects of EphB4 CAR‐T cells on lung adenocarcinoma cell lines were assessed. EphB4 and EphA2 positivity in lung adenocarcinoma samples was 93% and 100%, respectively. EphB4 CAR‐T cells were successfully generated with high CAR positivity and a high proportion of naïve/stem cell memory‐like T cells. In the antigen stimulation assay, the cells responded in an antigen‐specific manner after stimulation with both EphB4 and EphA2 proteins. In the co‐culture experiment, EphB4 CAR‐T cells significantly suppressed the growth of all four lung adenocarcinoma cell lines compared to mock‐T cells. In vivo, mice treated with EphB4 CAR‐T cells displayed a significantly lower tumor burden than those treated with either CD19 CAR‐T cells or phosphate‐buffered saline. Additionally, mice treated with EphB4 CAR‐T cells survived significantly longer than those in the other groups. In conclusion, ligand‐based EphB4 CAR‐T cells are bispecific, targeting both EphB4 and EphA2. Furthermore, these cells exert significant antitumor effects against lung adenocarcinoma.

Supramolecular RNAi with multifunctional siRNA nanostructures.

Development of a spatially defined 3D in vitro coculture construct modelling pancreatic cancer-associated cachexia.

The infiltration of regulatory T cells (Tregs) in lung adenocarcinoma (LUAD) is associated with a poor prognosis. The present study aimed to explore the potential function of Treg marker genes in prognosis and immunotherapy using the transcriptome profiles of five LUAD cohorts sourced from public databases. Among them, the single-cell dataset GSE131907 was employed to identify cell types in lung cancer tissues and to identify Treg markers. The prognostic Treg markers were screened using univariate Cox and Least Absolute Shrinkage and Selection Operator regression analyses. Subsequently, a prognostic model was constructed and assessed using Kaplan-Meier and receiver operating characteristic curves. Furthermore, the effect of prognostic Treg markers on clinical characteristics, the immune microenvironment and tumor mutation burden (TMB) were evaluated. In addition, the function of genes strongly correlated with the prognostic risk score were investigated using Spearman and functional enrichment analysis. Finally, the genes expression levels were assessed using reverse transcription-quantitative PCR (RT-qPCR) and western blotting. By analyzing the single-cell data, 13 Treg markers [centromere protein M (CENPM), pituitary tumor-transforming gene 1 protein, interleukin 1 receptor type 2, baculoviral IAP repeat containing 3, glucocorticoid induced 1 (GLCCI1), melanoma-associated antigen H1 (MAGEH1), CD5, cytokine inducible SH2 containing protein, zinc finger protein 101 (ZNF101), Ikaros family zinc finger protein 4 (IKZF4), ankyrin repeat and SOCS box protein 2, zinc finger CCCH-type containing 12D and C-C motif chemokine receptor 6] were identified as prognostic features. The prognostic model constructed using these 13 genes revealed that the high-risk group had a poorer prognosis than the low-risk group. Moreover, it was demonstrated that the risk score could be an independent prognostic factor affecting the prognosis of patients with LUAD. Additionally, the high-risk group had a lower ESTIMATE score, higher TMB score and lower T cell receptor richness than the low-risk group. Finally, RT-qPCR and western blotting showed that the expression levels of CENPM, ZNF101, MAGEH1 and IKZF4 were significantly altered in cancer tissues compared with the adjacent normal tissues. In conclusion, a reliable prognostic model based on 13 Treg markers was developed. The comprehensive characterization of the Treg markers of LUAD may help to monitor the prognosis and provide new strategies for LUAD treatment.

Colorectal cancer (CRC) remains one of the most prevalent and lethal malignancies worldwide, highlighting the urgent need for developing effective treatments. Molecular hybridization is a promising strategy for identifying new bioactive compounds. This study focused on designing and synthesizing a novel series of benzofuran-pterostilbene hybrid molecules. These compounds were successfully obtained, and their structures were elucidated by spectroscopic analysis. In addition, the activity of the hybrids was evaluated against colorectal adenocarcinoma cells. After the treatments, hybrids 6d and 6e exhibited the highest activity, with GI50 values of 11.93 ± 2.38 µM and 4.74 ± 0.38 µM, respectively, suggesting antiproliferative effects and measurable cytotoxicity under the tested conditions. Additionally, Hoechst 33342 fluorescence imaging revealed chromatin condensation and nuclear fragmentation, along with a diffuse DiOC₆ fluorescence pattern relative to the control, suggesting a form of programmed cell death, which was further supported by flow cytometric analysis showing an increased proportion of hypodiploid cells following propidium iodide staining. In parallel, wound-healing assays demonstrated impaired migration and cytotoxic effects that affected cell viability and structural integrity. Molecular docking simulations showed that compounds 6d and 6e bind strongly to mutant p53, CDK4, and PARP-1 proteins, which, in turn, may explain at the molecular level the in vitro cytotoxic effect of these compounds in SW480 colon cancer cells. Lastly, pharmacokinetic and toxicological modelling suggests that hybrids 6d and 6e possess optimal biopharmaceutical profiles with no major safety concerns. All these findings highlight the potential of the benzofuran-pterostilbene scaffold, with compounds 6d and 6e emerging as strong candidates for further evaluation against colorectal cancer.

Effective T-cell activation requires both antigen recognition and co-stimulatory signaling. However, tumor cells typically lack the necessary co-stimulatory molecules, rendering T-cell receptor engagement with tumor antigens insufficient to induce potent anti-tumor immune responses. To overcome this limitation, we converted tumor cells into professional antigen-presenting cell (APC)-like cells by equipping them with the co-stimulatory molecule CD80. Electroporation of melanoma and lung cancer cell lines with CD80-encoding mRNA significantly increased their ability to stimulate tumor-antigen-specific T cells, as indicated by increased activation markers and cytokine secretion. Most importantly, CD80 expression converted melanoma cells into APC-like cells capable of priming and expanding naïve T cells that recognize endogenous tumor antigens. To enable clinical application, we subsequently used poly-(beta aminoester) (pBAE) polymers, which efficiently encapsulate mRNA into nanometric polyplexes. As previously reported, pBAE nanoparticles (NPs) efficiently penetrate plasma membranes and escape endosomal degradation both in vitro and in vivo. Here, we evaluated pBAE NPs encapsulating GFP-encoding mRNA in human cell lines derived from melanoma, lung cancer, leukemia, adenocarcinoma, uveal melanoma, and Merkel cell carcinoma. The pBAE NPs effectively transfected adherent tumor cells, and tumor spheroids in vitro and in ovo. When delivering CD80 mRNA, NPs achieved expression rates of more than 50% in most cell lines, which lasted for at least 72 h. Here as well, nanoparticle-mediated CD80 expression enabled melanoma cells to prime and expand naïve T cells recognizing endogenous tumor antigens. Importantly, T cells primed by CD80-expressing tumor cells were also capable of killing tumor cells that had not been transfected. In line with these findings, T cells stimulated with untransfected tumor cells showed high levels of checkpoint receptors CTLA-4 and PD-1, whereas the expression was significantly lower when using CD80-transfected tumor cells. This proof-of-concept study indicates that conversion of tumor cells into professional APC-like cells with CD80 mRNA is feasible and results in potent anti-tumor CD8+ T-cell responses. With the use of mRNA-loaded pBAEs, the prerequisites for a future application in vivo are given. Therefore, the application of pBAE NPs encapsulating CD80 mRNA to specifically transfect tumor cells presents a new promising strategy for the in vivo treatment of various cancers.

Background and Purpose Quercetin , a widely distributed flavonoid in medicinal plants, exhibits potent anti-cancer effects across various malignancies. However, its pharmacological activity in gastric cancer remains insufficiently characterized. To investigate the anti-proliferative, pro-apoptotic, anti-migration, and anti-tumor effects of Quercetin against gastric cancer using in vitro (human gastric adenocarcinoma cell line [AGS], human gastric carcinoma cell line [MKN-45]) and in vivo xenograft models. Materials and Methods Cell viability, apoptosis, migration, and invasion assays were performed following Quercetin treatment (0–200 µM). A nude mouse xenograft model was induced by subcutaneous inoculation of MKN-45 cells, followed by oral Quercetin treatment (50 or 100 mg/kg/day). Tumor growth, inflammatory cytokines, and oxidative stress biomarkers were evaluated. Results Quercetin reduced the viability of AGS and MKN-45 cells in a dose-dependent manner, with IC 50 values of 46.3 and 52.7 µM, respectively. Apoptosis significantly increased under Quercetin treatment, while migration and invasion were markedly inhibited. In vivo, Quercetin suppressed tumor growth by up to 57.3% compared with controls and significantly reduced serum tumor necrosis factor-alpha, interleukin-6, interleukin-1β, and improved oxidative stress biomarkers. Conclusion Quercetin exerts strong therapeutic potential against gastric cancer by inhibiting proliferation, migration, and tumor progression, supporting its development as a natural product-based anti-cancer candidate.

mRNA sequencing analysis suggested that ubiquinone oxidoreductase subunit A10 (NDUFA10) may function as a key downstream effector of circRAPGEF5 in lung adenocarcinoma (LUAD) progression; however, the underlying mechanism remains unclear. This study aimed to explore the expression and clinical significance of NDUFA10 in LUAD cells and its role in cell proliferation and metastasis. Variations in NDUFA10 mRNA and protein expression between LUAD and adjacent normal tissues were assessed using the Cancer Genome Atlas Program (TCGA) and Tumor Immune Estimation Resource (TIMER) databases. These findings were subsequently validated in LUAD cell lines via Reverse Transcription Quantitative PCR (RT-qPCR). Further analyses included clinical correlation, Receiver Operating Characteristic (ROC), survival analysis, and Gene Set Enrichment Analysis (GSEA) of NDUFA10. The functional role of NDUFA10 was investigated in vitro at the cellular level. NDUFA10 mRNA expression was significantly upregulated in pan-cancer analyses and specifically in LUAD. Its expression levels correlated with clinical parameters, including metastasis (M stage) and patient gender. Elevated NDUFA10 expression demonstrated a negative correlation with overall survival (OS) and first progression (FP), and was associated with the best efficacy of anti-PD-1/PD-L1 antibody therapies. GSEA indicated significant enrichment of NDUFA10-related genes in pathways involving WNT and Notch signaling, as well as the TGFβ pathway regulating the epithelial-mesenchymal transition (EMT). In vitro, knockdown of NDUFA10 significantly suppressed LUAD cell proliferation, colony formation, migration, and invasion compared to control cells. This study focuses on the role of NDUFA10 in lung adenocarcinoma (LUAD). Through database screening and experimental verification, it was found that NDUFA10 is upregulated in LUAD. Its high expression is associated with advanced N stage (N2/N3), high pathological stage (III/IV), poor therapeutic effect and advanced age, suggesting its potential as a biomarker for poor diagnosis and prognosis. Further analysis indicates that NDUFA10 is associated with immune infiltration and involves pathways such as neuroactive ligand-receptor interaction and chemokine signaling. In vitro experiments have confirmed that knocking down NDUFA10 can inhibit the proliferation and metastasis of lung adenocarcinoma cells. This study provides a preliminary basis for the function and mechanism of NDUFA10 in LUAD. In the future, we will further explore how it affects tumor progression through the WNT/Notch/TGFβ pathway or ROS/ energy metabolism. Silencing NDUFA10 suppresses LUAD cell proliferation, migration, and invasion. The study suggested that circRAPGEF5 regulated NDUFA10 to facilitate the progression of LUAD.

We assessed whether Tridax procumbens (TP) extracts could be used therapeutically against pancreatic cancer and remain nontoxic to normal cell types. The crude extract from TP (CETP) was fractionated using hexane, dichloromethane, and ethyl acetate to obtain fractions (NHF, DCMF, and EAF, respectively). The pancreatic ductal adenocarcinoma cell line (PANC-1) was cultured with (10, 20, 50, 100, and 250 μg/mL) dimethyl sulfoxide (DMSO) (control), CETP, and CETP-fractions for 24 or 48 h. As a normal cell type, we cultured E11.5d mouse pancreatic explants for five days before treating with the test samples (20 μg/mL) in DMSO for a further 48 h. Cytotoxicity assays (MTT and Live-Dead) were conducted, and the expression of cellular biomarkers, such as vimentin, Ki-67, p53, p21, and caspase-3, was evaluated. DCMF elicited PANC-1 cell death (IC50 = 23.1 μg/mL) compared to CETP (IC50 = 114.2 μg/mL). There were significant elevations in p53 (2.8-fold), caspase-3 (2.9-fold), catalase (4.0-fold), p21Cip1/Wap-1 (4.4-fold), and ALP (5.0-fold) proteins in DCMF-treated cells compared to control. DCMF significantly suppressed PNA and GST-pi (2.3-fold), Ki-67 (2.7-fold), and vimentin (10.8-fold) in PANC-1 cells relative to control. Also, DCMF induced Bcl-2 perinuclear staining and cytoplasmic translocation of APC in treated cells. Phenotype morphogenesis was observed in DCMF-treated embryonic pancreas with immunopositivity for insulin, vimentin, and amylase (1.3-fold), cytokeratin-7 (1.5-fold), PNA (1.9-fold), and glucagon (2.8-fold). In conclusion, Tridax procumbens appear to exert its anticancer effect via induction of apoptosis, antioxidant enzymes, and suppression of growth, proliferation, and invasiveness in PANC-1 cells without any observable toxicity on the normal embryonic pancreas.

Canine pulmonary adenocarcinoma (cPAC) lacks well-validated systemic treatment options, motivating preclinical identification of potential therapeutic candidates. To explore potential therapeutic candidates, we first performed a screening of an FDA-approved drug library using a cPAC cell line, which identified bortezomib-a proteasome inhibitor-as a potential agent. Guided by this result, we focused subsequent investigations on proteasome blockade and evaluated the antitumor activity and mechanisms of bortezomib together with a second-generation proteasome inhibitor, carfilzomib-alone and in combination with standard cytotoxics-in three cPAC cell lines (CLAC, HDC, and LuBi). Both agents reduced cell proliferation and cell viability in a dose-dependent manner, with proteasome target engagement (ubiquitinated protein accumulation), G2/M arrest, sub-G1 increase, and caspase-3 cleavage. Pan- and selective caspase inhibition partially rescued viability, suggesting contributions from both intrinsic and extrinsic apoptosis, together with caspase-independent mechanisms. We also observed activation of the JNK-c-Jun pathway and increased Bim expression, indicating the induction of cellular stress responses and apoptosis. Drug-interaction analysis showed reproducible synergy between carfilzomib and carboplatin in HDC and LuBi, whereas combinations with vinorelbine were mainly additive or antagonistic effects. Overall, proteasome inhibitors exert multifaceted cytotoxic effects in cPAC through proteotoxic stress, cell-cycle arrest, and caspase-associated apoptosis. In particular, the combination of carfilzomib and carboplatin may represent a promising therapeutic strategy. Further invivo and clinical investigations are warranted to evaluate therapeutic efficacy, safety, and pharmacokinetics of proteasome inhibitors in cPAC.