Lysophosphatidic acid (LPA) is a small bioactive phospholipid which plays an important role during embryonic development and promotes developmental potential of in-vitro-produced (IVP) embryos in several species, including sheep and pigs. In bovines, LPA accelerates IVP blastocyst formation through the Hippo/YAP pathway. However, other LPA effects and its potential receptors during bovine embryo development are less clear. In this study, we used enzyme-linked immunosorbent assay (ELISA) to assess the presence of LPA in bovine oviductal fluid and determine cell apoptosis in embryos after LPA stimulation by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and quantitative reverse transcription polymerase chain reaction (qRT-PCR). We further evaluated potential receptors of LPA through molecular docking, RNA-seq data analysis and quantitative RT-PCR. LPA was found to be present in oviductal fluid. An increase in total cell number and a decrease in apoptosis levels were detected in day 7 blastocysts after LPA treatment. Among eight LPA receptors (LPARs), GPR87 and LPAR2 showed the highest affinity with LPA and their transcripts were expressed in embryos after the 16-cell stage in RNA-seq and qRT-PCR analysis. However, only the expression of LPAR2 was significantly increased in day 6 blastocysts after LPA stimulation, indicating its potential role in LPA-mediated signaling pathways. Our data highlight the positive effects of LPA on embryos and enrich information of related signaling mediators of LPA during embryonic development.

Prostate cancer remains a leading cause of cancer-related morbidity and mortality among men globally, with limited therapeutic options for advanced and metastatic disease. The therapeutic potential of natural compounds has attracted increasing attention in cancer treatment. Lanatoside C (Lan C), a cardiac glycoside derived from Digitalis lanata, has demonstrated promising anticancer activity across various cancer types. However, its role and mechanisms in prostate cancer remain underexplored. In this study, evidence shows that Lan C significantly inhibits the proliferation of prostate cancer cells, as demonstrated by reduced cell viability, suppressed colony formation, and G2/M cell cycle arrest. Additionally, Lan C promotes apoptosis and inhibits the migration and invasion of prostate cancer cells. Mechanistically, transcriptomic analysis identified differentially expressed genes, which were further validated at both the mRNA and protein levels. Our findings suggest that Lan C exerts its effects by modulating the TNF/IL-17 signaling pathway, influencing the tumor microenvironment and regulating key processes involved in tumor progression, immune response, and apoptosis.

Clinacanthus nutans is a valuable traditional medicinal plant that contains enriched active compounds such as triterpenoids and flavonoids. Understanding the accuulation process of these secondary metabolites in C. nutans requires exploring gene expression regulation under abiotic stresses and hormonal stimuli. qRT-PCR is a powerful method for gene expression analysis, with the selection of suitable reference genes being paramount. However, reports on stably expressed reference genes in C. nutans and even across the entire family Acanthaceae are limited. In this study, we evaluated the expression stability of 12 candidate reference genes (CnUBQ, CnRPL, CnRPS, CnPTB1, CnTIP41, CnACT, CnUBC, CnGAPDH, Cn18S, CnCYP, CnEF1α, and CnTUB) in C. nutans across different tissues and under abiotic stresses and MeJA treatment using three programs (geNorm, NormFinder, and BestKeeper). The integrated ranking results indicated that CnUBC, CnRPL, and CnCYP were the most stably expressed genes across different tissues. Under abiotic stress conditions, CnUBC, CnRPL, and CnEF1α were the most stable, while under MeJA treatment, CnRPL, CnEF1α, and CnGAPDH exhibited the highest stability. Additionally, CnRPL, CnUBC, and CnEF1α were the most stable reference genes across all tested samples, whereas CnGAPDH was the least stable. CnRPL, consistently ranking among the top three most stable genes, may therefore serve as an ideal reference gene for qRT-PCR analysis in C. nutans. To further validate the selected reference genes, we assessed the expression of two key biosynthetic genes, CnPAL and CnHMGR. The results confirmed that using the most stable reference genes yielded expression patterns consistent with biological expectations, while using unstable reference genes led to significant deviations. These findings offer valuable insights for accurately quantifying target genes via qRT-PCR in C. nutans, facilitating investigations into the mechanisms underlying active compound accumulation.

Lysine, butyric acid, and zinc play important roles in skin homeostasis, which involves aging, inflammation, and prevention of skin barrier disruption. This bioactivity spectrum is not replicated by any one topical compound currently in use. Our purpose in this study was to characterize a novel compound, zinc dibutyroyllysinate (ZDL), consisting of zinc with lysine and butyric acid moieties. We used RNA-seq to evaluate its effect on gene expression in a full-thickness skin model. We show that lysine alone has minimal effects on gene expression, whereas ZDL had greater transcriptional bioactivity. The effects of ZDL included an increased expression of genes promoting epidermal differentiation and retinol metabolism, along with a decreased expression of microphthalmia-associated transcription factor (MITF) and other melanogenesis genes. These effects were not replicated by an alternative salt compound (i.e., calcium dibutyroyllysinate). ZDL additionally led to a dose-dependent increase in skin fibroblast extracellular matrix proteins, including collagen I, collagen IV, and prolidase. Loss of melanin secretion was also seen in ZDL-treated melanocytes. These results provide an initial characterization of ZDL as a novel topical agent. Our findings support a rationale for the development of ZDL as a skincare ingredient, with potential applications for diverse conditions, involving melanocyte hyperactivity, pigmentation, inflammation, or aging.

Antimicrobial resistance is spreading rapidly throughout the world. The food chain can be one of the routes of transmission for microorganisms containing drug-resistance genes and thus serve as a channel for their transmission. Environmental stress and methods of preventing the spread of microorganisms trigger adaptive responses in bacterial cells. The aim of the present study was to determine the effect of the stress induced by sub-inhibitory concentrations (SICs) of cadmium chloride and benzalkonium chloride on antibiotic resistance and the expression of selected virulence factors in Staphylococcus isolates from food. The study was conducted on strains of the species S. epidermidis, S. heamolyticus, S. saprophyticus, and S. aureus. The values of the minimum inhibitory concentration against erythromycin, tetracycline, and oxacillin were determined before and after the incubation of the tested strains under stress conditions. The ability to form biofilm and slime production was also investigated. The expression levels of the genes responsible for antibiotic resistance (blaZ, tetK, tetM, ermB, and mecA) and virulence (eno) were conducted using Real-Time PCR. The MIC values of the antibiotics tested against the strains analyzed were found to be elevated in the presence of SICs of benzalkonium chloride and cadmium chloride. Furthermore, the intensity of biofilm production was also increased. SICs of benzalkonium chloride induced the expression of the tetM, tetK, mecA, and blaZ genes in 75%, 66.6%, 33.3%, and 40% of the isolates tested, respectively. Similar treatment with cadmium chloride induced the expression of the same genes in 75%, 100%, 66.6%, and 40% of the strains. In both cases, the expression of the ermB gene was reduced in 100% of the isolates. The eno gene was found to be overexpressed in 66.6% of the strains following benzalkonium chloride stress, and in 100% of the strains following cadmium chloride stress. These findings suggest that in Staphylococcus spp. strains, changes in the expression of the genes encoding antibiotic resistance and virulence factors may occur in response to the applied stress factors. The results indicate the possibility of selecting more resistant and virulent strains due to the use of too low concentrations of disinfectants, which emphasizes the need to use appropriate inhibitory doses of disinfectants in the food industry.

Drought is one of the main adverse factors affecting the growth and development of wheat. The molecular regulation pathway of Strigolactone (SLs or SL),which induces drought resistance in wheat, needs to be further clarified. In this study, SL and Tis (Strigolactone inhibitor) were sprayed on leaves to clarify the changes in wheat drought resistance and their effect on antioxidant enzyme activity, photosynthesis and other metabolic processes. However, 20 kinds of DOF transcription factors were identified by transcriptome metabolome association analysis, and they were highly enriched on chromosome 2. Moreover, the proline, glycosides, indoleacetic acid, betaine, etc., in wheat are the key factors affecting the change in the drought resistance of wheat. The study initially revealed the mechanism of the involvement of DOF in the SL regulation pathway and revealed its impact on different metabolites of wheat, thus providing a theoretical reference for the subsequent molecular verification and breeding of excellent drought-resistant varieties.

Vitamin D is a fat-soluble vitamin that has a protective role in colorectal cancer. Several studies have identified the association between vitamin D and changes in DNA methylation in different types of tumours. Dickkopf-1 (DKK1) inhibits the Wnt/β-catenin signalling pathway, and 1,25(OH)2D3 can induce DKK1 expression in colorectal cancer. However, whether 1,25(OH)2D3 can affect DKK1 expression by regulating DNA methylation in colorectal cancer is not known.

Lung cancer remains the leading cause of cancer-related mortality worldwide, necessitating the development of new treatment strategies. Arachidonic acid (ARA), a polyunsaturated fatty acid, shows promise in cancer therapy due to its potential anti-tumor effects, although its role in lung cancer remains unclear. This study investigated the effects and underlying mechanism of ARA on A549 and NCI-H1299 lung cancer cells. In vitro assays were used to assess cell viability, apoptosis, colony formation, lipid droplet formation, and changes in cellular lipid content. ARA dose-dependently suppressed cell viability, facilitated apoptosis, and suppressed colony formation in both lung cancer cell lines. Network pharmacology analysis was performed to identify potential gene targets and pathways, uncovering 61 overlapping genes between ARA and lung cancer-related targets, with mitogen-activated protein kinase 1 (MAPK1) emerging as a key gene. Enrichment analyses suggested that the effects of ARA might be mediated through lipid metabolism and the extracellular signal-regulated kinase (ERK)/peroxisome proliferator-activated receptor gamma (PPARγ) signaling pathway. In both lung cancer cell lines, ARA treatment inhibited lipid droplet formation and decreased the cellular lipids. Immunoblotting further confirmed that ARA treatment significantly increased ERK phosphorylation while reducing PPARγ and fatty acid synthase (FASN) protein levels. In vitro experiments using GW9662, a PPARγ antagonist, confirmed that inhibiting lipid droplet formation impairs lung cancer cell viability and promotes apoptosis. Furthermore, in vivo experiments demonstrated that ARA significantly reduced tumor size and weight in a lung cancer xenograft model, further validating its anti-tumor effects. The potential of ARA as a therapeutic agent for lung cancer might involve lipid metabolism and relevant signaling pathways. A future study exploring the full therapeutic potential of ARA and underlying mechanisms in lung cancer is needed.

Metabolic reprogramming plays a pivotal role in the development and progression of tumors. Tumor cells rely on glycolysis as their primary energy production pathway and effectively utilize biomolecules generated by the pentose phosphate pathway (PPP) for efficient biosynthesis. However, the role of 6-phosphogluconate dehydrogenase (6PGD), a crucial enzyme in the PPP, remains unexplored in esophageal squamous cell carcinoma (ESCC). In this study, we observed a significant upregulation of 6PGD expression in ESCC tissues, which correlated with an unfavorable prognosis among patients. The experiments demonstrated that knockdown of 6PGD induces oxidative stress and suppresses ESCC cell proliferation. Mechanistically, this is achieved through AMPK activation and subsequent inhibition of downstream mTOR phosphorylation. Moreover, physcion has been found to inhibit 6PGD activity and exert its anti-ESCC effect via the AMPK/mTOR pathway. Subsequently, we conducted both in vitro and in vivo experiments to validate the anticancer efficacy of combining metformin, an AMPK activator, with physcion. The results demonstrated a significantly enhanced inhibition of ESCC growth. This study elucidates the impact of 6PGD on ESCC cell proliferation along with its underlying molecular mechanisms, highlighting its potential as a therapeutic target for ESCC. Furthermore, we investigated a novel approach for improved anti-tumor therapy involving physcion and metformin. These findings will contribute new insights to clinical treatment strategies for ESCC while providing a theoretical foundation for developing molecular targeted therapies.

The treatment of gastric cancer remains challenging, with immunotherapy serving as a critical component of the holistic approach to its treatment. The results of this study indicated that statins could decrease the serum levels of interleukin-enhancing binding factor 3 (ILF3) and programmed cell death ligand 1(PD-L1) in GC patients and improve their prognosis. Functional experiments demonstrated that simvastatin induced ferroptosis by inhibiting ILF3 in GC cells and enhanced the killing effect of activated CD8+ T cells on GC cells. The CUT&Tag assay revealed that, mechanistically, simvastatin inhibited ILF3 expression by reducing the acetylation level at residue site H3K14 in ILF3. Next-generation sequencing and Kyoto Encyclopedia of Genes and Genomes analysis revealed that ILF3 regulated PD-L1 expression through the DEPTOR/mTOR signaling pathway. Overall, simvastatin induced ferroptosis in GC cells by inhibiting ILF3 expression while promoting the activation of CD8+ T cells to augment antitumor immune responses, thereby facilitating synergistic immunotherapy.

Glioblastoma (GBM) is the most lethal primary brain tumor with intra-tumoral hierarchy of glioblastoma stem cells (GSCs). The heterogeneity of GSCs within GBM inevitably leads to treatment resistance and tumor recurrence. Molecular mechanisms of different cellular state GSCs remain unclear. Here, we find that classical (CL) and mesenchymal (MES) GSCs are enriched in reactive immune region and high CL-MES signature informs poor prognosis in GBM. Through integrated analyses of GSCs RNA sequencing and single-cell RNA sequencing datasets, we identify specific GSCs targets, including MEOX2 for the CL GSCs and SRGN for the MES GSCs. MEOX2-NOTCH and SRGN-NFκB axes play important roles in promoting proliferation and maintaining stemness and subtype signatures of CL and MES GSCs, respectively. In the tumor microenvironment, MEOX2 and SRGN mediate the resistance of CL and MES GSCs to macrophage phagocytosis. Using genetic and pharmacologic approaches, we identify FDA-approved drugs targeting MEOX2 and SRGN. Combined CL and MES GSCs targeting demonstrates enhanced efficacy, both in vitro and in vivo. Our results highlighted a therapeutic strategy for the elimination of heterogeneous GSCs populations through combinatorial targeting of MEOX2 and SRGN in GSCs.

Staphylococcus aureus is one of the most common pathogens causing widespread infections. It has been demonstrated that thiazolidinone derivative (TD-H2-A), a small molecule compound that targets WalK protein through high-throughput screening, exerts antibacterial and anti-biofilm effects on S. aureus. In this study, we further ascertained the impact of TD-H2-A on biofilms at different stages. The phosphorylation assay and RNA sequencing were carried out to elucidate the underlying mechanism. The results revealed that TD-H2-A inhibited WalK autophosphorylation, implying that the antibacterial effect of TD-H2-A may be achieved by inhibiting the activity of WalK. The transcriptome analysis showed that TD-H2-A treatment induced 994 differentially expressed genes (DEGs), of which, 481 were upregulated and 513 were downregulated. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that 43 among 58 genes involved in ribosome synthesis were upregulated, and the transcript levels of the genes responsible for membrane transport were altered significantly. According to our research, TD-H2-A has an antibacterial mechanism with multitarget and multipathway. This study provided new ideas for the development of new drug target screening against S. aureus infections.

Vitamin C (VitC) has elicited considerable interest regarding its potential role in cancer therapy; however, its effects on tumor immunity remain unclear. In colorectal cancer (CRC), although anti-PD-1/PD-L1 therapies demonstrate promise, their efficacy is still constrained. Our prior research demonstrated that VitC can inhibit tumor growth by suppressing the Warburg effect. This study aims to explore the effects of high-dose VitC on PD-L1 expression in CRC, focusing on its underlying mechanisms and potential for enhancing immunotherapy. We found that VitC inhibits aerobic glycolysis in HCT116 cells while also downregulating PD-L1 expression. Further investigations indicated that this process is mediated by VitC's activation of AMPK, which downregulates HK2 and NF-κB, ultimately resulting in reduced PD-L1 expression and increased T cell infiltration. Notably, we observed that VitC and the PD-L1 monoclonal antibody atezolizumab exhibit comparable tumor-inhibiting abilities, and their combined use further enhances this efficacy. In conclusion, our results demonstrate that high-dose VitC activates AMPK, downregulates PD-L1 expression, mitigates immune evasion, and suppresses tumor growth. This provides a promising strategy for optimizing immunotherapy in CRC.

Lambda-Cyhalothrin (LCT), a potent insecticide, is renowned for its efficacy, rapid onset, and broad-spectrum insecticidal activity. With rising annual demand for LCT, environmental concerns have similarly intensified. Nevertheless, its impact on mammalian reproduction, specifically oocytes, and possible mitigation strategies, remains insufficiently understood. This research evaluated the effects of varying LCT concentrations (25, 50, 100 μM) on the in vitro maturation of porcine oocytes, revealing that LCT exposure markedly hindered oocyte maturation and subsequent developmental potential. Fisetin (FIS), a natural flavonoid found in numerous fruits and vegetables, exhibits robust antioxidant, anti-inflammatory, and neuroprotective properties. Notably, when different concentrations of FIS were incorporated into the culture medium of LCT-exposed oocytes, it mitigated LCT-induced toxicity. FIS supplementation significantly enhanced antioxidant capacity, improved endoplasmic reticulum (ER) and mitochondrial functions, and reduced apoptosis in comparison to the LCT-exposed group. Key markers, including the ER stress protein GRP78 (Control: 1.00 vs. LCT: 1.75 ± 0.08, P < 0.001; vs. LCT + FIS: 1.37 ± 0.07, P < 0.001) and autophagy marker LC3B (Control: 1.00 vs. LCT: 1.50 ± 0.32, P < 0.05; vs. LCT + FIS: 1.18 ± 0.16, P > 0.05), were significantly reduced in the FIS-treated oocytes. These results suggest that LCT compromises mitochondrial and ER function in oocytes through the upregulation of GRP78, while FIS supplementation effectively counteracts this toxicity, providing protection against LCT-induced damage. This study underscores FIS's potential as a protective agent in alleviating LCT-related reproductive toxicity.

VASCULAR SPECIFICITY FACTOR 1 (VSF-1) is a basic leucine zipper transcription factor identified in tomato (Solanum lycopersicum L.). VSF-1 regulates vascular-specific gene expression and is homologous to an Arabidopsis thaliana mechanical stress regulator, VIP1, but physiological roles for VSF-1 remain unclear. Here, we demonstrate that VSF-1 shuttles between the nucleus and the cytoplasm in response to hypo-osmotic stress. In Arabidopsis plants overexpressing the VSF-1-GFP fusion protein, VSF-1-GFP was mainly detected in the cytoplasm under unstressed conditions but in the nucleus under hypo-osmotically stressed conditions. VSF-1 contains three serine residues within HXRXXS motifs, which can serve as its phosphorylation and 14-3-3 protein-binding sites. In a transient gene expression system in Nicotiana benthamiana leaves, GFP-fused VSF-1 variants where those serine residues were replaced with alanine exhibited nuclear accumulation even under unstressed conditions. GFP-fused VSF-1 variants lacking those HXRXXS motifs also exhibited such nuclear accumulation. The VSF-1 variants lacking those HXRXXS motifs failed to interact with 14-3-3 proteins in a yeast two-hybrid system. These findings suggest that the nuclear accumulation of VSF-1 is triggered by hypo-osmotic stress through its dissociation from 14-3-3 proteins, similar to that of VIP1. The Arabidopsis VSF-1-GFP-overexpressing lines exhibited retarded germination and growth in the presence of mannitol, which can induce hyper-osmotic stress and repress nuclear accumulation of VSF-1. These results are consistent with phenotypes from VIP1-GFP-overexpressing lines in a previous study, indicating a conserved role for VIP1 and VSF-1 in regulating osmotic stress responses.

Background Gastric cancer is the world's leading tumor disease in terms of morbidity and mortality and is currently treated clinically with a comprehensive approach based on surgery. Studies have demonstrated the antitumor effects of neferine, but the anti-cancer mechanism for gastric cancer is not yet clear. Methods The Pubchem and Swiss TargetPrediction databases were searched to retrieve the targets of action of neferine. Meanwhile, relevant gene expression data were downloaded by means of the Gene Expression Omnibus(GEO) database to screen for differential genes and build a drug-disease network. The selected genes were analysed by bioinformatics analysis. Finally, gastric cancer treatment potential of neferine was determined through molecular docking. The molecular mechanism of neferine in the treatment of gastric cancer was verified by CCK8 assay, monoclonal assay, apoptotic and cycle assay, qRT-PCR and Western Blot. Results The results of network pharmacological analyses illustrate that the core genes are closely related to apoptosis, cell cycle, and cell proliferation. Through molecular docking, it was confirmed that neferine were closely related to key proteins. The results of in vitro experiments indicated that neferine could significantly inhibit the viability of gastric cancer cells, induce apoptosis of gastric cancer cells, and block the cell cycle of gastric cancer cells in the G0/G1 phase. Conclusion In summary, neferine inhibited the proliferation of gastric cancer cells through the CDK4/CDK6/CyclinD1 complex. This study provides a theoretical basis for the treatment of gastric cancer with neferine and an idea for the development of neferine for gastric cancer.

Cementoblasts and formation of new cementum are crucial for periodontal regeneration. The aim of this study is to investigate the effect of different graft materials on cementoblast's proliferation, mineralization and mineralized tissue-related gene expressions in-vitro. Immortalized mouse cementoblasts (OCCM-30) were treated with the media containing released components of graft materials (100 mg/ml ratio; waited in 5% FBS containing media for 3 days). Proliferation of the cells was evaluated using a real-time cell analyzer for 170 h and wound healing assay was performed to determine the migration of OCCM-30 cells. Total RNA was isolated on days 3 and 6, and biomineralization of the cementoblasts was assessed using von Kossa staining. mRNA expressions of bone sialoprotein (BSP), osteocalcin (OCN), collagen type I (COL-I), runt-related transcription factor2 (Runx2), and alkaline phosphatase (ALP) genes were examined using quantitative RT-PCR. While there was no significant difference at 72 h, all test groups showed significant reduction of cementoblast's proliferation at 96th and 120th h. All graft materials increased cell migration to the experimental wounded area. While Bio-Oss® showed significantly better effects on all mineralized tissue associated gene mRNA expressions on day 3 (p < 0.01), Nanobone® upregulated Runx2 (p < 0.01) and BSP (p < 0.05), Emdogain® induced OCN (p < 0.05) and Runx2 (p < 0.05). Endobon® upregulated Runx2 only on day 3 and 6 (p < 0.05). The most prominent increase in mineralized nodule formation was observed in Nanobone® (p < 0.05). In conclusion, released components of all graft materials have positive effects on the cementoblast's functions while Bio-Oss® might be preferable for gene expressions. Nanobone® has superiority for the biomineralization of the cementoblasts. The interaction of the graft materials and the cementoblasts is critical for the formation of new cementum required for periodontal regeneration.

Our study aims to investigate the potential of artesunate (ART) in improving learning and memory function by down-regulating NLRP3 and consequently affecting pyroptosis levels in the brains of rats with cerebral small vessel disease (CSVD). Initially, Sprague-Dawley (SD) rats were randomly assigned to five groups: the solvent sham operation group, solvent model group, low-dose ART (ARTL) group, medium-dose ART (ARTM) group, and high-dose ART group (ARTH). CSVD rat models were established through bilateral common carotid artery occlusion (BCCAO). Subsequently, the rats were further divided into four groups: the empty plasmid control group (shNC) and three groups receiving NLRP3-shRNA interference plasmids (shNLRP3-1, shNLRP3-2, shNLRP3-3). We recorded animal behaviors and stained nerve cell changes. Hippocampal expression levels of Caspase-1, cleaved caspase-1, IL-18, IL-1β, GSDMD-N, β-actin, and NLRP3 were evaluated in each group. Our findings revealed that ART ameliorated cognitive dysfunction and brain tissue injury in CSVD rats. Moreover, expression levels of cleaved caspase-1, IL-18, IL-1β, GSDMD-N, and NLRP3 in the hippocampus were significantly reduced in the shNLRP3 group, resulting in improved cognitive function in these rats. These results suggest that NLRP3 could be a potential therapeutic target in CSVD development in rats, and modulating its expression might mitigate pathological alterations associated with CSVD. Subsequently, lipopolysaccharide (LPS) was injected into the tail vein, and inflammatory factors in peripheral blood of rats were found to be increased, suggesting that the level of intracranial NLRP3 was increased. In addition, MWM experiment showed that after the increase of NLRP3 expression, the repair effect of ART on learning and memory dysfunction was weakened. ART may enhance cognitive impairment in CSVD rats by downregulating NLRP3 expression in the brain, thereby inhibiting neuronal cell pyroptosis in the hippocampus.

Cuproptosis is a form of copper-dependent non-apoptotic cell death. Cancer cells that prefer to use aerobic glycolysis for energy generation are commonly insensitive to cuproptosis, which hinders its application for cancer treatment. Epigallocatechin gallate (EGCG) possesses diverse pharmacological activities. However, the association between EGCG and cuproptosis has not been studied.

Type 2 diabetes is a risk factor for colorectal cancer (CRC) development and progression. However, metformin-treated diabetic CRC patients tend to have better clinical outcomes than those managed by other means. To better characterize the molecular underpinnings of metformin's protective effects, we performed a targeted transcriptomic analysis of primary CRC tissue samples (n = 272). A supervised learning algorithm pinpointed molecular features that discriminate between metformin-treated and diet-controlled diabetic CRC samples, as well as those that discriminated between non-diabetic samples based on their five-year overall survival status. Our results show downregulation of TMEM132 in metformin-treated samples (p = 0.05) and non-diabetics with good clinical outcomes (p = 0.05) relative to diet-controlled and non-diabetics with poor survival, respectively. Furthermore, upregulation of SCNN1A is observed in metformin-treated samples (p = 0.04) and non-diabetics with good clinical outcomes (p = 0.01) relative to diet-controlled samples and those with poor clinical outcomes, respectively. We also show that the antiapoptotic protein sFas is downregulated in metformin-treated samples relative to diet-controlled samples (p = 0.005). These findings suggest a role for the unfolded protein response in mediating metformin-related CRC-protective effects by enhancing apoptosis and suggest the investigation of these proteins as targets for novel CRC therapies.