Colon cancer death toll due to metastasis is expected to rise. Extracellular vesicles are signalling molecules which can regulate communication between cells and either inhibit or promote cancer metastasis. Extracellular vesicles from adipose mesenchymal stem cells of Macaca fascicularis were obtained from previous research. Various concentrations of extracellular vesicles (5 μg/mL, 10 μg/mL, and 20 μg/mL) were applied to colon cancer cell line WiDr in a transwell invasion assay. CXCR3 gene expression analysis was done using RT-qPCR. Application of extracellular vesicles successfully inhibited colon cancer invasion with decrease of number of invasive cells with increase in concentration. There was no statistical difference between cells treated with 10 μg/mL and 20 μg/mL of extracellular vesicles. Colon cancer cells treated with extracellular vesicles show upregulation of CXCR3A and CXCR3B gene expression compared to negative control. Extracellular vesicles inhibit colon cancer cell invasion through upregulation of CXCR3B expression.

ELK1/NOL3/GRP78 axis regulates proliferation and stemness in TP53-mutant colon cancer by enhancing adaptive endoplasmic reticulum stress.

PKM2 inhibition by genistein induces metabolic disruption and apoptosis in colon cancer cells

Anticancer drug-treated fibroblasts modulate colon cancer cell behavior through lysophosphatidic acid (LPA) receptor-mediated signaling.

Bis-(phenylselenide) based turn-on fluorescent probe for rapid, perceptible and selective detection of superoxide anion in human colon cancer cells

Optimization of lens size and ion source parameters for gas chromatography/mass spectrometry-based metabolomics using hydrogen as a carrier gas.

Selenoproteins are critical regulators of redox homeostasis, protein folding, and metabolism, and their dysregulation has been implicated in cancer biology. Among them, selenoprotein F (SELENOF) has been reported to be tumor suppressive, whereas the RNA-binding protein EIF4A3, a component of the exon junction complex, has been implicated in post-transcriptional repression of selenoproteins. The regulatory and clinical significance of this interaction in colorectal adenocarcinoma (COAD) remains unclear. We performed an integrative analysis of transcriptomic data from The Cancer Genome Atlas (TCGA), proteomic data from the Clinical Proteomic Tumor Analysis Consortium (CPTAC), and patient tissue microarrays. Western blotting, qRT-PCR, and immunofluorescence staining were used to examine SELENOF, GPX1, and EIF4A3 expression in colon cancer cell lines and tumor tissues. Correlation, regression, and survival analyses were conducted, and pathway enrichment was assessed using gene set enrichment analysis (GSEA) of RNA and proteome correlation profiles. Motif discovery and translational efficiency analyses were performed to identify 3'-UTR features associated with EIF4A3 repressive activity. SELENOF and EIF4A3 showed inverse, stage-dependent protein expression patterns in COAD in the CPTAC cohort. Survival analyses demonstrated that SELENOF alone was not prognostic but acquired significance in EIF4A3-high tumors, where low SELENOF was associated with poor outcomes. Motif analyses identified enriched 3'UTR elements in SELENOF, suggesting that EIF4A3 represses translation through non-SECIS motifs positioned near canonical SECIS elements. Our findings explore a novel EIF4A3-SELENOF regulatory axis in colorectal cancer. SELENOF acquires conditional prognostic significance only in the context of elevated EIF4A3, highlighting the importance of molecular interaction specificity in biomarker discovery.

Design and synthesis of novel 1,2,4-triazolobenzene sulfonamide derivatives as selective CDK1 inhibitors with potent in vivo anticancer efficacy.

Effect of TU-100 on Colorectal Liver Metastasis in Mouse Model of MASH.

Syndesmos (SDOS), which regulates cytoskeletal organization via interaction with syndecan-4 in fibroblasts, was under-expressed in highly metastatic HCT116 colon cancer cells compared to weakly metastatic HT29 cells. SDOS overexpression decreased migration and proliferation of HCT116 cells, while siRNA-mediated knockdown of SDOS increased these activities in HT29 cells. SDOS overexpression in HCT116 cells promoted epithelial cell-like morphology, enhanced cell-cell and cell-ECM junction formation, and increased expression of epithelial markers. SDOS also elevated adhesion receptors, such as E-cadherin and syndecan-4, and their intracellular anchoring proteins. Notably, SDOS specifically interacted with E-cadherin and β-catenin, enhancing their interaction at cell-cell junctions in both HCT-SDOS and HT29 cells, suggesting a direct role of SDOS in adherens junction formation. In colon cancer tissue samples, SDOS expression was lower compared to adjacent normal tissues and lower SDOS levels were associated with poor post-progression survival in patients. In vivo, SDOS overexpression inhibited tumor growth in a HCT116 xenograft mouse model. Silencing SDOS in CT26-luc cells via siRNA increased tumor growth in the cecum and liver metastasis in an orthotopic model. These findings suggest that SDOS plays a critical role in suppressing colon cancer development and metastasis by maintaining epithelial architecture and promoting cell-cell and cell-ECM adhesion.

BackgroundSulfated polysaccharides (SPs) from red algae have attracted considerable interest due to their antioxidant and anticancer properties. Gracilaria corticata is a rich source of sulfated polysaccharides with notable antioxidant and anticancer potential.ObjectivesThis study aimed to optimize the extraction process of SPs from G. corticata, determine their chemical and structural features, and evaluate their antioxidant and anticancer activities.Materials and methodsThe SPs were extracted from G. corticata using ultrasound-assisted extraction. The chemical composition, molecular weight (230 kDa), and structure of the extracted SPs were characterized using standard biochemical assays and nuclear magnetic resonance (NMR) spectroscopy. Antioxidant capacity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. Cytotoxicity was assessed against HT-29 colon cancer cells and normal CCD-841 cells via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis induction was further examined through Annexin V-FITC/PI staining and gene expression analysis of apoptosis-related markers.ResultsUltrasound-assisted extraction was optimized at 12 min, 800 W power, and a solvent-to-biomass ratio of 30 mL/g, yielding 34.8% SPs. The SPs contained 84.4% carbohydrates, 8.7% sulfate, and 2.55% protein, with galactose identified as the predominant sugar. Molecular weight was determined to be 230 kDa. Antioxidant assays demonstrated dose-dependent activity, reaching 81.92% (DPPH) and 86.34% (ABTS) radical scavenging. SPs inhibited HT-29 cell viability with an IC₅₀ of 171 μg/mL and a selectivity index (SI) of 3.48. Apoptosis was induced via increased intracellular reactive oxygen species (ROS) and altered expression of key genes, including upregulation of Bax, P53, and Caspase 3, and downregulation of BCL2.ConclusionsSPs extracted from G. corticata, show strong antioxidant and selective anticancer activity through ROS-mediated apoptosis. These findings support their potential use as natural therapeutic agents in pharmaceutical development.

Disruption of proteostasis is a defining feature of cancer and other chronic diseases. The AAA+ ATPase VCP/p97 (valosin containing protein) is a key regulator of proteostasis by disassembling ubiquitinated substrates for degradation. VCP overexpression supports cancer cell survival and correlates with poor prognosis, promoting the development of VCP inhibitors as anti-cancer agents. However, the molecular basis for cancer-selective vulnerability of VCP inhibition remains unclear. Here, we demonstrate that allosteric VCP inhibition triggers cell- type specific macroautophagy/autophagy through dynamic reorganization of organelle contact sites. In human umbilical vein endothelial cells (HUVECs), VCP inhibition induces adaptive autophagy through coordinated reorganization of plasma membrane (PM)-ER-mitochondria contacts. Controlled opening of the mitochondrial permeability transition pore (mPTP) releases calcium into the cytosol, activating AMP-activated protein kinase (AMPK) and TFEB pathways, collectively enhancing autophagic flux and sustaining endothelial survival. Critically, calcium-activated kinase inhibitor or calcium chelators blocked VCP inhibitor-induced autophagy in HUVECs, confirming calcium signaling as the central mediator of adaptive autophagy. In contrast, HCT116 colon cancer cells fail to maintain calcium homeostasis under VCP inhibition, leading to mitochondrial calcium overload, defective autophagy, and cell death. Together, our findings identify organelle contact reorganization and calcium homeostasis as key determinants of cell fate under conditions of proteotoxic stress, revealing how VCP inhibition selectively suppresses tumor progression while preserving vascular integrity that could enhance drug delivery and reduce tumor hypoxia.

Secreted PEBP4 Promotes Colorectal Cancer Progression via Regulation of the TGF-β Signaling Pathway.

Ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, has emerged as a promising therapeutic target for colorectal cancer (CRC). However, the precise mechanisms by which CRC cells evade ferroptosis remain incompletely understood. Cysteine redox modification, typically catalyzed by cysteine oxidases, is a key regulatory factor governing protein structure and function. Quiescin sulfhydryl oxidase 1 (QSOX1), a Golgi-localized sulfhydryl oxidase known to promote various physiological functions, has an uncharacterized role in ferroptosis. Therefore, this study investigated the effects of QSOX1 on ferroptosis sensitivity in colorectal cancer. We utilized a comprehensive set of analytical techniques to elucidate the mechanisms of QSOX1 in CRC ferroptosis resistance. We assessed cell proliferation, colony formation, and sensitivity to ferroptosis inducers (Erastin and RSL3) in CRC cells. Levels of reactive oxygen species (ROS), intracellular Fe2+, and redox metabolites were measured by biochemical assays and flow cytometry. RNA sequencing (RNA-seq) and untargeted metabolomics were performed using QSOX1-deficient HT-29 cells. The interaction between QSOX1 and SLC7A11 was confirmed via coimmunoprecipitation, immunofluorescence, and nonreducing gel electrophoresis. Cystine uptake and glutamate release assays were used to assess SLC7A11 function. Tumor xenografts were generated to assess the in vivo sensitivity of colon cancer cells to ferroptosis inducers. QSOX1 was significantly upregulated in CRC tissues and promoted CRC cell proliferation in vitro and in vivo. Knockdown of QSOX1 sensitized CRC cells to ferroptosis inducers, whereas QSOX1 overexpression conferred resistance. Mechanistically, QSOX1 enhanced redox homeostasis and GSH metabolism in CRC cells. QSOX1 interacted with and facilitated redox modification of SLC7A11 at cysteine 158 via its thioredoxin motif (C449-C452) in the ERV/ALR domain. This oxidative regulation enhanced SLC7A11 membrane localization, cystine uptake, and glutamate release, thereby boosting intracellular GSH synthesis and suppressing ferroptosis. QSOX1 promotes ferroptosis resistance in CRC cells through redox-dependent post-translational modification of SLC7A11, enhancing its activity and promoting GSH synthesis. The QSOX1-SLC7A11 axis represents a potential therapeutic target to overcome ferroptosis resistance in CRC.

Altered lipid metabolism is a potential targetable metabolic vulnerability in colorectal cancer (CRC). Fatty acid synthase (FASN), the rate limiting enzyme of de novo lipogenesis, is an important regulator of CRC progression, but the FASN inhibitor TVB-2640 showed only modest efficacy in reducing tumor burden in pre-clinical studies, suggesting combination strategies might be required to prolong patient survival. Here, by using samples from a window trial of TVB-2640 treatment in CRC patients, we found that FASN inhibition induced DNA damage but impaired the DNA damage response (DDR). In colon cancer cell lines and patient-derived organoids, FASN inhibition potentiated chemotherapy-induced double-strand DNA breaks (DSBs) and apoptotic cell death by altering histone acetylation levels. In addition, FASN inhibitor treatment blocked DDR by decreasing ATM expression and CHK2 phosphorylation. Mechanistically, FASN inhibition attenuated activation of the DDR pathway by attenuating BRCA1 and ATM recruitment to γ-H2AX foci in an acetylation-dependent manner. Moreover, FASN inhibition mediated DNA repair deficiency induced synthetic lethality with PARP inhibition in CRC cells. Importantly, combining FASN inhibition with the chemotherapeutic drug irinotecan synergistically decreased xenograft tumor growth and delayed tumor relapse, which was potentiated by the PARP inhibitor olaparib as maintenance treatment. Taken together, this study describes a therapeutic strategy in which FASN inhibitors can be utilized to delay tumor recurrence after chemotherapy, which is a major challenge in patients with CRC.

Metastatic recurrence represents the major clinical challenge in early-stage lung cancer after curative surgery. Here, we investigated the role of circulating extracellular vesicles and particles (EVPs) in promoting formation of pre-metastatic niches (PMNs). Plasma-derived EVPs were obtained by ultracentrifugation from pre-surgery blood samples of patients with poor prognosis. Heavy-smokers cancer free individuals were used as control. EVP were characterized following MISEV guidelines. Functional experiments were carried out in vitro in 2D and 3D-bioprinted models as well as in vivo. EVPs from patients with early relapse show distinct molecular profiles, characterized by elevated levels of miR-29a and complement protein C4a. These EVPs preferentially target endothelial cells inducing a pro-inflammatory condition with upregulation of VCAM1 and CXCL1. In turn, endothelial modulation stimulated fibroblast activation and promoted neutrophils recruitment supporting PMNs formation. Mechanistically, we demonstrate that miR-29a and C4A act synergistically through SPARC down-modulation promoting cancer cell colonization. Preconditioning of mouse lungs using EVPs from patients with poor prognosis increased metastatic growth of human tumor cells, which was inhibited by miR-29a blockade. Circulating EVPs could be novel prognostic biomarkers and key players in PMN formation offering new targets to reduce relapses in lung cancer.

Anoctamin 5 (ANO5), a member of the anoctamin/transmembrane protein 16 family, has been implicated in various cancers; however, its role in colon cancer (CC) remains unclear. We examined ANO5 expression in CC cell lines and human tumor tissues and investigated its functional significance. ANO5 knockdown using small interfering RNA markedly suppressed proliferation, migration, and invasion of HCT116 and LoVo cells, while inducing G0/G1 cell-cycle arrest and apoptosis. Microarray and validation analyses revealed activation of the TP53-BAX-CASP3 apoptotic pathway and regulation of CDKN1A and CDK2, suggesting that ANO5 modulates both apoptosis and G1/S checkpoint progression. Immunohistochemistry of tumors from 200 patients with stage II/III CC demonstrated high ANO5 expression in 52% of cases. High expression was significantly associated with poorer outcomes, with 5-year relapse-free survival (62.6% vs. 87.3%, p < 0.001) and overall survival (74.0% vs. 94.8%, p < 0.001) markedly worse in the high-expression group. Multivariate analyses confirmed high ANO5 expression as an independent prognostic factor. Collectively, ANO5 promotes tumor cell growth, survival, and motility, and its high expression predicts unfavorable clinical outcomes, indicating its potential as a prognostic biomarker and therapeutic target in CC.

Skeletal muscle atrophy is a devastating and defining feature of cancer cachexia that reduces quality of life, treatment tolerance, and survival, but cannot be prevented or reversed by current management strategies. Ursolic acid is a natural dietary compound that has been shown to inhibit atrophy-associated changes in skeletal muscle mRNA expression in rodents and dogs, leading to beneficial changes in skeletal muscle structure and function. We hypothesized that dietary supplementation with ursolic acid might help support skeletal muscle mass and function during cancer. To test this hypothesis, we investigated ursolic acid's effects in five in vivo mouse models of cancer cachexia that are driven by pancreatic, colon, and lung cancer cells of mouse and human origin. We found that dietary supplementation with ursolic acid has broad-spectrum effects towards cancer-induced skeletal muscle atrophy, significantly preserving muscle mass in all five cancer cachexia models. Ursolic acid's positive effects on muscle mass and muscle fiber size led to significant improvements in grip strength and muscle tetanic force, persisted in the presence of chemotherapy, and were not associated with discernable changes in food intake or tumor growth. Ursolic acid appeared to generate its beneficial effects in skeletal muscle by acting directly on muscle cells, inhibiting catabolic effects of tumor-derived secreted factors, and inhibiting > 90% of cancer-induced changes in skeletal muscle mRNA expression. These results strongly nominate ursolic acid as a promising potential nutritional approach for supporting muscle mass and function in individuals with cancer.

This study aimed to investigate the mechanism by which allicin, a bioactive compound from garlic, exerts antitumor effects, specifically focusing on its role in regulating the long non-coding RNA UCA1 via the induction of autophagy. Experiments were conducted using human colorectal cancer cell lines HCT-116 and HT-29. Cell proliferation was measured with a CCK-8 kit, apoptosis was analyzed by flow cytometry, cell invasion was assessed using Transwell chambers, and migration was evaluated via a scratch wound assay. Autophagic structures were examined under a transmission electron microscope. The expression level of lncRNA UCA1 was determined by quantitative reverse transcription polymerase chain reaction (RT-qPCR), and the expression of proteins associated with autophagy was detected by western blot analysis. Compared with the control group, allicin treatment significantly inhibited cancer cell proliferation, increased the rate of apoptosis, and reduced capabilities for invasion and migration. Furthermore, allicin downregulated the expression of lncRNA UCA1 in a concentration-dependent manner and simultaneously increased the number of autophagosomes. All these effects were statistically significant (P < 0.05). However, when cells were co-treated with an mTOR activator, the antitumor effects of allicin and the increase in autophagosomes were significantly counteracted (P < 0.001). Allicin inhibited colon cancer cell activities through the induction of cellular autophagy and the subsequent regulation of lncRNA UCA1 expression.

The development of IL-N8 and its magnetic micelle-forming derivative MIL-N8 offers a promising advancement in nanostructured carriers for efficient hydrophobic drug delivery. The structural validation for MIL-N8 was obtained through Raman, EPR, and NMR spectroscopy, which suggested successful synthesis and molecular integrity of both IL-N8 and MIL-N8 compounds. Furthermore, the thermogravimetric analysis demonstrated that MIL-N8 exhibits better stability than its precursor. Due to the structural attributes of MIL-N8 (containing both hydrophilic and hydrophobic moieties), it readily self-assembles into uniform nanostructures, such as micelles, at the critical aggregation concentration (CAC). This was further characterized through confocal microscopy using ANS as a fluorescent probe and visualized using TEM and FESEM imaging. These nanomicellar structures enabled the efficient encapsulation of the hydrophobic anticancer drug, quercetin (QCT), with high loading and sustained release behavior adhering to the Korsmeyer-Peppas model, which suggests diffusion-controlled transport through aggregated micellar layers. Furthermore, biological evaluation using SW-480 colon cancer cells demonstrated a remarkable enhancement in the anticancer activity of QCT-encapsulated and delivered via MIL-N8, moreover, in comparison to free QCT, the QCT-MIL-N8 formulation produced substantially lower IC50 values and pronounced dose and time-dependent effects. QCT-MIL-N8 formulation demonstrated increased apoptotic features, including nuclear condensation, fragmentation, and an increase in AO/EtBr-positive cells, as well as elevated intracellular ROS levels, that further supported oxidative stress-driven cell death as the predominant mechanism. Collectively, MIL-N8 nanomicelles emerge as a highly effective, low-toxicity delivery platform that significantly improves the therapeutic potential and controlled release of QCT for anticancer applications.