Metformin reduces the incidence of breast cancer in patients with obesity and type 2 diabetes. However, our knowledge of the effects of metformin on breast cancer recurrence is limited. Within the randomized double-blind placebo-controlled phase II trial MetBreCS, we examined changes in breast tissue from breast cancer survivors with BMI > 25 kg/m2 after treatment with metformin. To identify metformin-regulated signaling pathways, we integrated the transcriptomic, metabolomic and steroid hormone profiles using bivariate and functional analyses. We identified MS4A1, HBA2, MT-RNR1, MT-RNR2, EGFL6 and FDCSP expression to be differentially expressed in breast tissues from metformin-treated postmenopausal women. The integration of transcriptomic and metabolomic profiles revealed down-regulation of immune response genes associated with reduced levels of arginine and citrulline in the metformin-treated group. The integration of transcriptomic and steroid hormone profiles showed an enrichment of steroid hormone biosynthesis and metabolism pathways with highly negatively correlated CYP11A1 and CYP1B1 expression in breast tissue from postmenopausal metformin-treated women. Our results indicate that postmenopausal breast cancer survivors treated with metformin have specific changes in breast tissue gene expression that may prevent the development of new tumors.Trial registration: MetBreCs trial is registered at European Union Clinical Trials Register (EudraCT Protocol # 2015-001001-14) on 07/10/2015.

Tumor budding (TB) and poorly differentiated clusters (PDCs) are features of infiltrative growth patterns and powerful independent prognostic factors in colorectal cancer (CRC), yet the underlying biological mechanisms behind their role in CRC invasion is less understood. The aim of this study was to investigate the molecular background and prognostic role of tumor cluster size at the invasive margin (IM) of CRC, and determine whether a biological continuum between TB and PDCs exists. Using a combination of spatial transcriptomic and immunohistochemical (IHC) techniques, we demonstrated a biological continuum from larger to smaller tumor clusters, with TB possessing greater invasive potential than PDCs. We deployed artificial intelligence on a cohort of 1134 Stage I-III CRC resections to automatically detect nearly 400,000 isolated tumor cells/clusters of any particular size across the IM. We determined that 2-celled clusters were the most abundant feature at the IM, and the simultaneous assessment of TB and PDCs yielded a prognostic performance stronger than either independently. Our study provides a deeper understanding of the mechanisms behind CRC invasion while improving risk stratification for Stage I-III CRC.

Platelets are essential anucleate blood cells that play pivotal roles in hemostasis, tissue repair, and immune modulation. Originating from megakaryocytes in the bone marrow, platelets are small in size but possess a highly specialized structure that enables them to execute a wide range of physiological functions. The platelet cytoplasm is enriched with functional proteins, organelles, and granules that facilitate their activation and participation in tissue repair processes. Platelet membranes are densely populated with a variety of receptors, which, upon activation, initiate complex intracellular signaling cascades. These signaling pathways govern platelet activation, aggregation, and the release of bioactive molecules, including growth factors, cytokines, and chemokines. Through these mechanisms, platelets are integral to critical physiological processes such as thrombosis, wound healing, and immune surveillance. However, dysregulated platelet function can contribute to pathological conditions, including cancer metastasis, atherosclerosis, and chronic inflammation. Due to their central involvement in both normal physiology and disease, platelets have become prominent targets for therapeutic intervention. Current treatments primarily aim to modulate platelet signaling to prevent thrombosis in cardiovascular diseases or to reduce excessive platelet aggregation in other pathological conditions. Antiplatelet therapies are widely employed in clinical practice to mitigate clot formation in high-risk patients. As platelet biology continues to evolve, emerging therapeutic strategies focus on refining platelet modulation to enhance clinical outcomes and prevent complications associated with platelet dysfunction. This review explores the structure, signaling pathways, biological functions, and therapeutic potential of platelets, highlighting their roles in both physiological and pathological contexts.

M2-polarized tumor-associated macrophages (TAMs) are a key factor contributing to the poor prognosis of pancreatic ductal adenocarcinoma (PDAC). While various factors within the tumor microenvironment (TME) drive their formation, the role of PDAC-derived exosomes in this process remains unclear. We aim to clarify the regulatory impacts of tumor-derived exosomes to TAMs. After the intratumoral injection to subcutaneous tumor of C57BL/6 mice, we demonstrated PDAC-derived exosomes exacerbate PDAC progression, accompanied with upregulated M2 phenotype of TAMs and unaffected proliferation signatures. Through intratumoral injection model and multi-Omics analyses, we identified CCT6A as a novel tumor-derived exosomal protein, bridging TAMs M2 polarization and PDAC prognosis. Co-culture with exosomes derived from CCT6Ahigh PDAC leads to greater M2 phenotype of TAMs via PI3K-AKT signaling. According to proteomics data, chemokines' abundance reduces over tenfold once exosomal CCT6A absence, including CXCL1, CXCL3, CCL20 and CCL5, whose interaction with CCT6A in PDAC cells was confirmed by interactomics data. Moreover, we found silencing CCT6A abrogated the antagonism effects of CD47 antibody immunotherapy. Our findings implied that the subunit of the T-complex protein Ring Complex (TRiC) CCT6A serves as a matchmaker during exosome-mediated chemokines transfer from PDAC to TAMs. Silencing CCT6A effectively sensitized PDAC to CD47 antibody immunotherapy in vivo.

Giant congenital melanocytic nevus (GCMN) is a RAS/RAF mutation-driven syndrome characterized by extensive melanocytic lesions, posing psychological challenges and a lifelong risk of malignancy. Existing treatments like surgical resection and laser therapy fail to fully remove lesions, and MAPK inhibitors show limited efficacy. This study identified a predominant population of senescent cells and a minority of proliferative cells in GCMN, necessitating dual-targeted strategies. We found that the anti-apoptotic protein BCL2 is expressed in both senescent and proliferative cells from GCMN patients with various gene mutations. Coexpression of P16 and BCL2 indicated a phenotype of growth arrest and cell survival. BCL2 inhibitors (BCL2i) showed significant cytotoxicity to GCMN cells in vitro. Hypopigmentation and GCMN cell clearance were observed in patient-derived xenograft models and in NrasQ61K-mutated and BrafV600E-mutated transgenic models following BCL2i treatment. Histology of regressed GCMN indicated extensive immune cell infiltration, suggesting immune involvement. Single-cell sequencing and immunostaining revealed that activated neutrophils formed extracellular traps, synergizing with BCL2i to treat GCMN. Neutrophil depletion and immunosuppression reduce treatment efficacy, highlighting the crucial role of the immune response post-BCL2i treatment. Long-term follow-up showed no recurrence, with neutrophils and T cells residing in the dermis, indicating memory immune reactions. These findings present a promising therapeutic strategy and underscore the translational potential of BCL2i in treating GCMN.

Neoadjuvant immunotherapy can induce pathologic complete response (pCR) in patients with localized deficient mismatch repair (dMMR)/microsatellite instability-high (MSI-H) tumors. The long-term outcomes of these patients are unknown, as is the clinical utility of measuring circulating tumor DNA (ctDNA). Follow-up was evaluated in patients enrolled in a phase II trial (NCT04082572) that evaluated the efficacy and safety of pembrolizumab in patients with localized dMMR/MSI-H tumors. The primary outcomes of this trial have previously been reported. 3-year EFS and OS rates were 80% (95% CI: 66% - 93%) and 94% (95% CI: 86% - 100%). Patients without detectable ctDNA after pembrolizumab had higher 3-year EFS and OS rates than patients with detectable ctDNA after pembrolizumab (3-year EFS 92% vs 20%; p < 0.001, 3-year OS 100% vs 80%; p < 0.001). Patients with colorectal cancer (CRC) who had undetectable ctDNA after pembrolizumab were more likely to have pCR compared to those with detectable ctDNA after pembrolizumab (91% vs 0%; p = 0.03). Patients with CRC who were managed non-operatively and had undetectable ctDNA after pembrolizumab had a higher 2-year EFS rate than patients with detectable ctDNA after pembrolizumab (100% vs 33%; p = 0.03). Pembrolizumab demonstrates long-term efficacy in patients with localized dMMR/MSI-H tumors.

The aberrant expression of S100A11 has been identified in various malignancies but its functional roles and underlying mechanisms in colorectal cancer (CRC) have not been fully elucidated. Therefore, this study was designed to investigate the expression of S100A11 and its functional significance in CRC, indicating that S100A11 is significantly upregulated and correlates with poor survival outcomes in CRC. Functionally, S100A11 knockdown in CRC cell lines inhibited cell proliferation, invasion, and migration, leading to decreased tumour growth and metastasis in vivo. Mechanistic investigations revealed that S100A11 promotes cell proliferation and invasion by suppressing cell senescence. In addition, USP14 interacts with and mediates S100A11 deubiquitination. More importantly, the overexpression of S100A11 was able to partially counteract the reduction in cell proliferation caused by the knockdown of USP14. In summary, the novel regulatory axis involving USP14 and S100A11 modulates the malignant biological behavior of CRC cells through inhibiting cell senescence, therefore the interaction between USP14 and S100A11 represents a promising therapeutic target in CRC.

Hepatocellular carcinoma (HCC) is an increasingly prevalent malignant neoplasm on a global scale. Circrna HIPK3 (circHIPK3) has been identified as playing a key role in HCC tumorigenesis and as a novel biomarker. In this study, we aimed at developing a sensitive and accurate method for the detection of circHIPK3 in low load plasma samples using a droplet digital PCR (ddPCR). We designed circHIPK3 gradient primers and probes and optimized the PCR system to improve performance. Then we assessed and compared the linearity and sensitivity of ddPCR and quantitative PCR (qPCR) using the circHIPK3 plasmid DNA as a template. Using these methods, circHIPK3 concentrations were quantitatively determined in 3 cell lines and 40 plasma samples to assess clinical stability. Within the plasmid concentration range of 31-36 copies/μl, the ddPCR exhibited a linear fitting equation of Y = 1.037X-0.1724 with R² value of 0.9940, which surpassed the corresponding R² value of 0.9877 for qPCR. Furthermore, the limit of blank (LOB) and limit of detection (LOD) for ddPCR were determined to be 0.157 copies/μl and 0.594 copies/μl, respectively, which were significantly lower than the LOD of qPCR (5.753 copies/μl). In clinical samples, ddPCR demonstrated a commendable correlation with qPCR, evidenced by a Kappa value of 0.677 (p < 0.05, 95% CI [0.503-0.851]) and an intraclass correlation coefficient (ICC) of 0.903 (95% CI [0.831-0.946]). Notably, ddPCR identified 11 positive samples that qPCR failed to detect. The ddPCR-based circHIPK3 liquid biopsy method emerged as a highly sensitive and accurate approach, lending itself well to clinical applications.

Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide, posing a significant threat to human health. Recently, gambogic acid (GA) has garnered attention for its anticancer properties in GC. However, it remains unclear whether GA can regulate other forms of cell death beyond apoptosis. In this study, we found that GA inhibited proliferation and induced ferroptosis in GC cells. Western blot analysis was employed to assess ferroptosis and endoplasmic reticulum (ER) stress-related proteins, as well as forkhead box A2 (FOXA2) expression. Additionally, malondialdehyde (MDA) and glutathione (GSH) levels were measured following GA treatment, and quantitative real-time polymerase chain reaction (RT-qPCR) was used to evaluate miR-1291 expression. Our findings revealed that GA treatment elevated reactive oxygen species (ROS) levels and promoted intracellular Fe[Formula: see text], MDA, and GSH accumulation. Furthermore, GA upregulated SLC7A11 and ferritin expression while suppressing glutathione peroxidase 4 (GPX4) in AGS and HGC27 cells, suggesting its role in ferroptosis induction. Notably, GA increased miR-1291 levels and downregulated FOXA2 expression. Subsequent analyses showed FOXA2 as a direct target of miR-1291. Functional experiments involving miR-1291 and FOXA2 knockdown or overexpression further suggested that the miR-1291/FOXA2 axis mediates ferroptosis. Finally, tumor xenograft models showed that GA effectively inhibited tumor growth by inducing ferroptosis. In conclusion, our study provides compelling evidence that GA induces ferroptosis in GC through the miR-1291/FOXA2 axis, highlighting its potential as a novel therapeutic strategy and preventive target for gastric cancer treatment.

Gastric cancer is a significant global health issue. Celastrol, a natural compound, has shown antitumor potential, but its molecular mechanism in gastric cancer remains unclear. In this study, we treated HGC-27 cells with celastrol and employed CCK8, colony formation, and Transwell assays, revealing its inhibitory effect on cell proliferation and migration. Flow cytometry assay results showed that celastrol could elevate the level of reactive oxygen species (ROS) in HGC-27 cells. By using the iron ion and malondialdehyde (MDA) detection kits, it was found that celastrol promoted the accumulation of iron ions (Fe[Formula: see text] in HGC-27 cells, increased the MDA content, and simultaneously decreased the glutathione (GSH) content. Additionally, Western blot analysis indicated that celastrol exerts an inhibitory effect on the expression of ferroptosis-marker proteins GPX4 and SLC7A11. PCR array and further experiments identified CERKL as a key factor, whose downregulation by celastrol was associated with enhanced ferroptosis. In vivo, celastrol inhibited tumor growth without affecting body weight or organ histology. Our findings suggest that celastrol may inhibit gastric cancer via CERKL-regulated ferroptosis, providing a potential therapeutic strategy.

N6-methyladenosine (m6A) modification, as a widespread and high-profile type of epigenetic modification, can dynamically and reversibly regulate the whole process of RNA metabolism. This modification governs RNA stability, splicing, and translation via m6A regulators, which are categorized as Writers, Erasers, and Readers. m6A modification also plays a critical role in the development of tumors. Disruptions in the homeostasis of m6A modification levels lead to dysregulation of m6A regulators. Consequently, these dysregulated regulators modulate the proliferation, migration, and invasion of tumor cells and interfere with the normal functions of suppressor genes and oncogenes. This article aims to comprehensively elucidate the specific regulatory impacts of m6A modification on tumor-related gene expression. It focuses on the regulatory mechanisms of m6A modification on mRNA stability. Additionally, it explores the influence of m6A on the molecular translation of key signaling pathways. Moreover, it investigates the indirect regulatory effects mediated by non-coding RNAs (ncRNAs), etc. The intention is to offer a novel analysis of the pathogenesis of cancer at a new level, and also provide a theoretical basis for finding new effective treatment methods.

Objective: To systematically investigate the association and regulatory mechanism between genetic variants in the binding region of pancreatic and duodenal homeobox 1 (PDX1) and pancreatic cancer susceptibility in the Chinese population. Methods: Chromatin immunoprecipitation sequencing (ChIP-seq) was performed using the human pancreatic cancer cell line BxPC-3 to identify and annotate genetic variants within the PDX1 binding region. A two-center case-control study was conducted, and logistic regression models were employed to analyze the association between PDX1-related variants and pancreatic cancer susceptibility. Functional experiments were performed to elucidate the molecular mechanisms of these genetic variants. Results: ChIP-seq analysis identified 1 608 PDX1 binding regions. SNPs within these regions were significantly enriched in susceptible areas of pancreatic cancer (P<0.001). The common variant rs154659, located within the most significant PDX1 binding peak, was further investigated. The multivariate logistic regression model showed that compared with individuals with TT genotype, individuals with CC genotype had a reduced risk of pancreatic cancer by 29.2% (OR=0.708, 95%CI: 0.589-0.850). Functional studies demonstrated that the rs154659[C] allele displayed higher relative luciferase activity than the rs154659[T] allele. Knockdown of PDX1 significantly attenuated the relative luciferase differences between the two alleles. Conclusion: Genetic variants in the PDX1 binding region are associated with pancreatic cancer risk. The rs154659 modulates pancreatic cancer susceptibility by specifically altering PDX1 binding activity.

Pathogenic variants in the BRCA1/2 genes significantly elevate risks of breast, ovarian, prostate, and pancreatic cancer. Clinical guidelines recommend cascade screening (CS) to identify at-risk family members and advance genetically targeted disease prevention. However, despite the benefits of CS, testing uptake remains suboptimal, particularly among male first-degree relatives (FDRs) of female BRCA1/2 carriers.

Pancreatic cancer has become one of the most challenging malignant tumors. This type of cancer shows a high mortality paralleling morbidity and a rather low 5-year survival rate. Early diagnosis of the disease is the key to prevent and treat the pancreatic cancer. Cellular free DNA (cfDNA) methylation detection is a promising non-invasive method in early cancer screening, diagnosis, and prognostic monitoring. However, there are still shortcomings in developing fast, sensitive, low-cost, and quantitative methods for detecting target methylated DNA. Herein, new transcription activator-like effectors (TALEs)-mediated electrochemical sensor by rolling circle amplification (RCA)-assisted "silver-link" crossing electrode was developed for detecting DNA methylation of Ras association domain family 1 isoform A (RASSF1A) tumor suppressor genes (R-5mC). The TALEs-Ni magnetic beads could specifically recognize R-5mC and bind to target methylated DNA to trigger the RCA and catalytic hairpin assembly (CHA), thereby duplexes (H1-H2) were generated. By introducing H3 probe to cause CHA amplification and Exo III-assisted cyclic amplification strategy, large amounts of H2 probes were generated to trigger a new round of RCA reaction, long crossing DNA strands were formed between the gaps of electrodes. The "silver-link" crossing electrode was formed by metallization of "gene-link" and target was detected by recording this change in conductivity. The limit of detection (LOD) was 0.5 fM. The proposed electrochemical sensor showed good stability, high sensitivity and low detected background signal in the R-5mC detection. In addition, the R-5mC detection of cfDNA in plasma samples of cancer patients was satisfactory. This strategy is of great significance in the early screening, treatment and prognosis monitoring of pancreatic cancer.

Genomic testing for prostate cancer (PCa) clinical management and hereditary cancer assessment has grown in clinical impact; however, challenges remain regarding optimal implementation and end-user confidence. The Decision-making, Experience, and Confidence In Determining Genomic Evaluation (DECIDE) survey was designed to collect information regarding utility and understanding of genomic testing from PCa health care providers, researchers, and stakeholders.

Patients with metastatic melanoma who progress on checkpoint inhibitors and BRAF-targeting drugs have limited therapeutic options. Up to one third of melanomas harbor at least one molecular aberration in the homologous recombination (HR) pathway, leading to HR deficiency.

In cancers with both high microsatellite instability (MSI-H) and BRCA1/2 mutations, BRCA1/2 mutations may be incidentally caused by MSI and represent passenger mutations versus drivers of those cancers. Reporting of these mutations without additional clarification may result in poly (ADP-ribose) polymerase (PARP) inhibitor therapy, where there is not likely true benefit. The purpose of this work was to identify BRCA1/2 passenger mutation hotspots that are secondary to MSI-H status rather than truly independent driving mutations.

Oligoprogressive disease (OPD) commonly occurs in patients with advanced EGFR mutation-positive non-small cell lung cancer (EGFR+ LC) on systemic therapy. While radiation therapy (XRT) to treat OPD can improve outcomes, the clinical and genomic predictors of benefit from local therapy for oligoprogression on osimertinib are unclear.

Glioma is a particularly lethal central nervous system tumor. Identifying the boundary between gliomas and normal tissues is difficult due to their infiltrative and invasive growth characteristics. This can result in the inevitable recurrence of the tumor after surgery. Preventing the residual tumor from growing or spreading is a major obstacle in treating gliomas. An earlier study suggested that hypotaurine could enhance the invasion of glioma cells while inhibiting the activity of demethylases. The hypotaurine synthesis-deficient U251 cell line usage showed a decrease in the cells' invasion capability. Analysis of gene expression profiles showed that reducing the activity of a critical enzyme in hypotaurine production, 2-aminoethanethiol dioxygenase (ADO), had a notable effect on the extracellular matrix-receptor interaction. Decreased intracellular ADO expression led to a significant increase in Wnt5a expression. Cells exposed to hypotaurine exhibited decreased levels of both intracellular Wnt5a protein and its corresponding mRNA. The observed characteristic was linked to increased methylation of the Wnt5a gene promoter, possibly due to hypotaurine's ability to inhibit demethylase enzymes. To sum up, the research showed that U251 cells lacking hypotaurine synthesis were susceptible to epigenetic changes, and Wnt5a seemed to function as a cancer inhibitor in this scenario. It would be beneficial to reevaluate this tumor suppressive effect in real tumor samples, which may contribute to the development of new glioma interference strategies.

Breast cancer remains a significant concern worldwide, with a rising incidence in Indonesia. This study aims to evaluate the applicability of risk-based screening approaches in the Indonesian demographic through a case-control study involving 305 women. We developed a personalized breast cancer risk assessment workflow that integrates multiple risk factors, including clinical (Gail) and polygenic (Mavaddat) risk predictions, into a consolidated risk category. By evaluating the area under the receiver operating characteristic curve (AUC) of each single-factor risk model, we demonstrated that they retained their predictive accuracy in the Indonesian context (AUC for clinical risk: 0.67 [0.61,0.74]; AUC for genetic risk: 0.67 [0.61,0.73]). Notably, our combined risk approach enhanced the AUC to 0.70 [0.64,0.76], highlighting the advantages of a multifaceted model. Our findings demonstrate for the first time the applicability of the Mavaddat and Gail models to Indonesian populations, and show that within this demographic, combined risk models provide a superior predictive framework compared to single-factor approaches.