Molecular Carcinogenesis最新文献

Our previous preclinical studies demonstrated that non-steroidal anti-inflammatory drugs (NSAIDs) such as aspirin and naproxen significantly inhibited prostate tumorigenesis in a TMPRSS2-ERG fusion-driven model compared to non-fusion models. Since TMPRSS2-ERG fusion-positive tumors display heightened inflammatory signaling and substantial immune infiltration, we hypothesized that the differential efficacy of NSAIDs may arise from their ability to remodel the tumor immune microenvironment. Accordingly, in the present study, we systematically profiled innate and adaptive immune-cell populations: F4/80⁺ macrophages, mast cells, neutrophils, CD3⁺ T cells, CD8⁺ cytotoxic T cells, FoxP3⁺ regulatory T cells, CD20⁺ B cells, IgKC⁺ plasma cells, and Granzyme B⁺ effector cells in highly infiltrated dorso-lateral prostate regions of TMPRSS2-ERG fusion-driven and non-fusion PCa models, with and without NSAID intervention. Our analyses revealed pronounced macrophage infiltration in TMPRSS2-ERG. Pten^flox/flox and Hi-Myc^{+/ -} model, which was further augmented by NSAIDs. Importantly, NSAID intervention shifted macrophage polarization toward an M1-like, pro-inflammatory state, contrasting with the M2-dominant phenotype characteristic of untreated tumors. NSAID treatment reduced mast cell density within the stromal compartment, suggesting suppression of mast cell-mediated tumor-promoting signals. In the fusion model, infiltration of total T cells and CD8⁺ cytotoxic T cells decreased following NSAID exposure, whereas FoxP3⁺ Tregs remained largely unaffected. Both models showed increased B-cell infiltration independent of NSAID efficacy, and no clear correlation was observed between plasma-cell presence and treatment response. Collectively, our findings offer new insight into NSAID-mediated immunomodulation in TMPRSS2-ERG fusion-driven PCa; however, further in-depth immune subtyping and spatial mapping could fully delineate the immunological mechanisms driving NSAID responsiveness.

Early diagnosis of acute promyelocytic leukemia (APL), driven by PML-RARA oncoprotein, is vital for survival, as delays can cause fatal coagulopathy without prompt therapeutic intervention of all-trans retinoic acid and arsenic trioxide. Although APL is diagnosed using microscopy, immunophenotyping, and FISH/PCR for PML-RARA, morphological overlap with acute myeloid leukemia (AML) -M5 and AML-M2 complicates identification. In resource-limited settings, unavailability of real time -quantitiative PCR (RQ-PCR) or delays in FISH/qualitative RT-PCR results increase the risk of missed or late diagnosis with fatal outcomes. Podoplanin (PDPN), a glycoprotein overexpressed in APL cells, interacts with platelets to mediate thrombosis. We evaluated PDPN expression, regulation, and diagnostic potential in APL. PDPN mRNA (RQ-PCR) and surface protein (flow cytometry) were significantly higher in APL than non-APL AML (p < 0.0001), consistent with TCGA-LAML and BEATAML1.0 datasets. Sensitivity and specificity were 80.7% and 71.43% by RQ-PCR, and 92.86% and 100% by flow cytometry, respectively. Chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) and TGF- β1 stimulation confirmed SMAD binding and PDPN upregulation. Pharmacological inhibition of TGF-β1 ligand using luspatercept reduced SMAD phosphorylation and PDPN expression, indicating TGF-β/SMAD transcriptionally regulates PDPN. Additionally, ELISA-based serum profiling showed significantly elevated TGF-β1 levels in APL patients compared to non-APL AML (p < 0.0001). These findings identify PDPN overexpression as a downstream consequence of TGF-β/SMAD signaling and highlight its potential as a diagnostic biomarker for APL.

Epithelial splicing regulatory protein 2 (ESRP2) is a splicing regulator specific to epithelial cell types. Multiple studies have found that its expression is abnormal in various tumors, influencing their occurrence, development, or prognosis. Our previous research indicated that down-regulating ESRP2 can suppress the proliferation of breast cancer (BC) cells, specifically the MCF-7 line. To delve deeper into the role of ESRP2 in BC cells, we investigated its impact on the migration of BC cells in vitro and the underlying molecular mechanisms. The outcomes of the in vitro scratch assay and Transwell assay initially confirmed that ESRP2 hinders the migration of both MCF-7 and MDA-MB-231 cells, and that reducing ESRP2 expression enhances their migratory capacity. We demonstrated that the down-regulation of ESRP2 can boost the migration ability of MCF-7 cells, and increase the mRNA expression of epithelial-mesenchymal transition (EMT) transcription factor ZEB2 and related markers N-cadherin and Vimentin. This suggests that ESRP2 plays a potential regulatory role in inhibiting EMT transcription program. Additionally, results from RNA sequencing and agarose electrophoresis gel experiments predict that down-regulating ESRP2 may promote exon skipping of the ENAH gene by modulating the alternative splicing of genes associated with cell migration, driving the shift of MCF-7 cells from an epithelial to a mesenchymal phenotype. Our research reveals a novel mechanism by which ESRP2 affects BC metastasis through post-transcriptional regulation. ESRP2 may present as a promising biomarker in combating BC cell migration by targeting EMT.

Glioblastoma (GBM) represents an exceedingly malignant and invasive type of brain tumor, known for its significant tendency to recur and its unfavorable clinical outcome. Despite significant advancements in research and therapeutic strategies, the survival rates for patients with GBM remain low. Synthesis of 1,4,5,6,7,8-hexahydropyridine [4,3-d] pyrimidine (PPM) has emerged as a promising avenue for the development of novel therapeutic agents with potent anti-glioma efficacy and minimal toxicity. Although previous studies have demonstrated the ability of PPM to inhibit HepG2 cell proliferation and suppress liver cancer, its potential anti-glioma effects and the underlying mechanisms remain largely unexplored. Using a comprehensive array of experimental approaches, this study was designed to elucidate both the therapeutic potential and underlying molecular mechanisms of PPM in GBM, including western blotting, flow cytometry, EdU assay, clone formation analysis, immunofluorescence, qRT-PCR, wound healing assay, Transwell migration/invasion assay, and a dual-luciferase reporter gene system. In vitro experiments demonstrated that PPM exhibited significant inhibitory effects on tumor cell proliferation, migration, and invasion. Furthermore, PPM treatment induced cell cycle arrest and promoted both apoptosis and autophagy, and its antitumor effects were mediated by the inhibition of autophagosome degradation. Mechanistically, PPM exerted its anti-glioma effects through the upregulation of miR-873-3p, which subsequently inhibited the PI3K/AKT/mTOR signaling pathway. Through the integration of proteomic profiling and bioinformatics approaches, PTBP1 was identified as a putative downstream target gene regulated by miR-873-3p. Suppression of miR-873-3p upregulates PTBP1 expression, thereby facilitating the progression and development of glioma cells. These findings suggest that PPM is a promising therapeutic candidate for GBM treatment and offer novel insights into glioma therapy. The identification of the miR-873-3p/PTBP1 axis as a potential therapeutic target provides a new direction for future clinical applications and GBM treatment strategies.

Hepatocellular carcinoma is a leading cause of cancer-related mortality worldwide, with metabolic syndrome emerging as a major risk factor. However, the molecular mechanisms underlying the association between metabolic syndrome and hepatocellular carcinoma progression are not fully understood. Here, we investigated the role of kisspeptin signaling in hepatocellular carcinoma progression under metabolic dysregulation. High-fat diet feeding significantly decreased hepatic kisspeptin receptor expression in mice. Integrated transcriptomic and metabolomic analyses revealed that kisspeptin primarily regulated glycolysis-related pathways. In a N-Nitrosodiethylamine-induced hepatocellular carcinoma mouse model, high-fat diet accelerated tumor progression accompanied by kisspeptin receptor downregulation. Treatment with kisspeptin-10 attenuated high-fat diet-promoted hepatocellular carcinoma progression and decreased the expression of key glycolytic enzymes HK, PFKM, and PKM2. In vitro studies using HepG2 cells further confirmed that kisspeptin-10 inhibited these glycolytic enzymes in a dose-dependent manner. The integration of transcriptomic and metabolomic data demonstrated that kisspeptin exerts broad inhibitory effects on metabolism, particularly glucose metabolism, also suggesting potential antitumor effect. Our results suggest kisspeptin as a potential therapeutic target for hepatocellular carcinoma in patients with metabolic syndrome.

Ovarian cancer (OC) is a leading cause of cancer-related mortality among females worldwide. Lysine demethylase 6A (KDM6A) plays a crucial role in multiple physiological and pathological processes. However, its role in ovarian carcinogenesis remains unclear. The expression of KDM6A and survival analysis in OC were assessed utilizing GEPIA and Kaplan-Meier plotter databases. The expression of KDM6A was evaluated immunohistochemically in tissue samples from 55 OC patients. The CCK-8, Colony formation, and Transwell assays were employed to assess the ability of OC cells in proliferation, migration, and invasion. Lung metastasis and subcutaneous tumor models were used to evaluate the function of KDM6A in vivo. RNA sequencing, Western blot, and quantitative polymerase chain reaction were conducted to investigate the molecular functions of KDM6A. A chromatin immunoprecipitation assay was employed to determine the effects of KDM6A on the promoters of ubiquitin-like protein interferon-stimulated gene 15 (ISG-15). KDM6A expression was downregulated in OC and associated with poor progression-free survival and overall survival. KDM6A inhibits OC cell proliferation, migration, and invasion in vitro. Xenograft models have also confirmed the antitumor role of KDM6A in OC growth and metastasis. The mechanistic study demonstrated that KDM6A exerted an antitumor effect in a histone-demethylase-dependent manner by epigenetically activating ISG-15 transcription. KDM6A, a functional tumor suppressor, is frequently downregulated in OC. The KDM6A-ISG-15 axis is critical in restraining OC malignancy and may serve as a potential molecular target for novel therapies.

Tertiary lymphoid structures (TLS) demonstrate prognostic significance and associations with immunotherapy response in gastrointestinal malignancies, though their regulatory mechanisms remain incompletely defined. The current understanding of TLS at single-cell resolution is limited. Here, we integrated single-cell and spatial transcriptomics with TLS-specific signatures to map spatial distributions and chemokine signaling within colorectal (CRC) and gastric cancer (GC) microenvironments. We identified significant enrichment of characteristic T cell and macrophage subsets in the TLS regions. Subpopulation analyses revealed distinct cellular interaction networks: CRC exhibited robust intercellular communication among effector CD8⁺ T cells, exhausted CD8⁺ T cells, tissue-resident CD8⁺ T cells, CD16⁺ monocyte-derived macrophages, C1QC⁺ macrophages, and SPP1⁺ macrophages. Conversely, GC featured pronounced interactions between interferon-stimulated gene-positive (ISG⁺) CD8⁺ T cells and ISG15⁺ macrophages. Further analyses suggest CD16⁺ monocyte-derived macrophages may recruit effector CD8⁺ T cells via the CXCL16-CXCR6 ligand-receptor pair in CRC, while ISG15⁺ macrophages may utilize dual CXCL16-CXCR6 and CXCL10-CXCR3 pairs to recruit ISG⁺ effector CD8⁺ T cells in GC. Our study uncovers spatially resolved, cancer-type-specific immune recruitment circuits within TLS, providing mechanistic insights into their functional organization and potential therapeutic targeting.

Linarin is a natural flavonoid glycoside that has extensive pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer effects. However, the functions of linarin in colorectal cancer have not been fully elucidated. The purpose of our study was to investigate the effect of linarin on colorectal cancer and delineate its potential molecular mechanisms. The malignant behavior of colorectal cancer cells were investigated utilizing colony formation, 5-ethynyl-2'-deoxyuridine, Annexin V/PI double staining, scratch, and transwell assays. The potential mechanism of linarin's anticancer activity was explored using network pharmacology analysis, molecular docking, qRT-PCR, immunohistochemical staining, western blot analysis, immunofluorescence, CHX chase assay, ubiquitination assay, and rescue experiments. In addition, mouse xenograft tumor models were used to confirm the role of linarin in colorectal cancer in vivo. We found that linarin inhibited the proliferative, migratory, and invasive abilities, but enhanced the apoptotic ability of colorectal cancer cells. Moreover, we also discovered that linarin could repress HIF-1α expression and HIF-1α/PD-L1 axis in LoVo and HCT-15 cells. Both HIF-1α and PD-L1 overexpression reversed the effect of linarin on the malignant behavior of colorectal cancer cells. Furthermore, linarin treatment significantly inhibited colorectal cancer tumor growth in vivo. In conclusion, linarin could inhibit the proliferative, migratory, and invasive capacity, but enhance the apoptotic ability in colorectal cancer cells through repressing the HIF-1α/PD-L1 axis.

Sphingosine-1-phosphate receptor 1 (S1PR1, also known as EDG1) is a G-protein-coupled receptor. Although there have been conflicting results reported, S1PR1 is generally accepted to be protumorigenic in many cancer types. However, S1PR1 has not been studied in thyroid cancer. This study aimed to investigate the biological activity of S1PR1 in thyroid cancer. S1PR1 protein levels were found to be higher in thyroid cancer tissues than adjacent normal tissues. Using an S1PR1-GFP construct, we showed that S1PR1 is localized in the cell membrane; however, when stimulated by sphingosine-1-phosphate, S1PR1 appeared to move inside in the cells. In functional studies, S1PR1 knock-out cells generated using the CRISPR/Cas9 system exhibited reduced S1PR1 activity, including colony formation, cell migration, and cell invasion. This was accompanied by the inhibition of STAT3, ERK1/2, and AKT kinase activity and cell adhesion protein expression. Furthermore, we screened several anticancer compounds to determine their effects on S1PR1 expression levels in thyroid cancer cells and found that quercetin significantly reduced S1PR1 protein levels in these cells. Overall, our results indicated that S1PR1 expression at the protein level has a positive relationship with thyroid cancer progression, as seen in other cancers. These data also suggest that quercetin is a potential anticancer drug that can target S1PR1-positive cells.

Hepatocellular carcinoma (HCC), the most common type of primary liver cancer, is characterized by a complex pathogenesis and high mortality rate. Currently, there is a lack of effective therapeutic agents for HCC. This study aimed to identify potential targeted therapeutic compounds for HCC and to validate their mechanisms of action through in vitro and in vivo experiments. Using the Connectivity Map database, we screened compounds capable of influencing the status of core genes and selected those predicted to be nontoxic for experimental validation. Our findings demonstrated that, within a certain concentration range (0-80 μM), Purvalanol A significantly inhibited the viability and proliferation of HCC cell lines Huh7 and Hepa1-6. Notably, the IC50 value for normal human liver cells (THLE-2) was much higher than that for HCC cells, indicating selective cytotoxicity. Purvalanol A also suppressed the migration and invasion abilities of HCC cells, induced G2/M cell cycle arrest, and promoted apoptosis. Additionally, Purvalanol A treatment downregulated the expression of the key gene CDK1 (cyclin-dependent kinases) and the antiapoptotic protein Bcl2 while upregulating the expression of p53, phosphorylated p53 (p-p53), and the p53 downstream proapoptotic protein Bax, moreover, the ratio of p-p53/p53 increased, indicating activation of the p53 pathway. In a mouse xenograft tumor model, Purvalanol A significantly inhibited subcutaneous tumor growth without causing noticeable toxicity to internal organs. In conclusion, the results of this study suggest that Purvalanol A exerts anti-HCC effects by activating the p53 pathway, making it a potential therapeutic compound for the treatment of hepatocellular carcinoma.