Exploring the mechanism by which triphenyl phosphate promotes malignant phenotypes in bladder and kidney cancer through MMP9 based on bioinformatics analysis and experimental validation
Jul 2026· Clinical and Experimental Medicine (Testo stampato)· Vol 26· 0 citations· 71 references
Medicine
TL;DR
TPP promotes the malignant phenotypes of bladder and kidney cancer via MMP9, which is validated by virtual knockout and in vitro experiments.
Abstract
Background Bladder and kidney cancer burden rises globally, with environmental toxicants driving their progression. Methods We integrated global epidemiological analysis, single-cell transcriptomics, cell–cell communication analysis, epithelial subclustering, pseudotime inference, toxicological target prediction, survival modeling, cross-cohort validation, single-cell virtual knockout, spatial transcriptomic deconvolution, molecular docking/dynamics, CETSA, and in vitro assays to define a shared molecular interface linking triphenyl phosphate (TPP) to bladder and kidney cancer. Results Both malignancies exhibited age- and SDI-associated burden patterns. Single-cell profiling identified conserved epithelial, stromal, and immune ecosystems, with tumor epithelial cells occupying central positions in intercellular communication networks. Epithelial subclustering revealed a reproducible EMT-high subcluster 4 in both cancers, which localized to a terminal-like pseudotime state and was associated with poor survival. Predicted TPP targets intersected with subcluster 4 signatures and converged on extracellular matrix organization, adhesion, and leukocyte transendothelial migration pathways. Integrative survival modeling and multi-cohort validation identified MMP9 as a robust prognostic candidate associated with tumor progression. Importantly, single-cell virtual knockout of MMP9 revealed convergent remodeling of proliferative, inflammatory, hypoxia-related, and stress-response programs across bladder and renal cancer epithelial cells, highlighting conserved regulatory circuitry. Spatial transcriptomics further localized MMP9 to macrophage- and fibroblast-enriched niches in both tumor types. Structural modeling and CETSA supported an interaction between TPP and MMP9. Experimentally, TPP upregulated MMP9 at both mRNA and protein levels in T24 and 786-O cells; higher concentrations reduced viability, whereas lower concentrations enhanced migration and clonogenic growth. Conclusions TPP promotes the malignant phenotypes of bladder and kidney cancer via MMP9, which is validated by virtual knockout and in vitro experiments.
Prostate cancer recurrence reflects molecular and histological heterogeneity, yet the cellular states harboring recurrence-associated signals and their potential environmental modifiers remain incompletely understood.
We integrated single-cell RNA sequencing, bulk transcriptomes with recurrence annotation, quantitative histopathology, cross-cohort survival modeling, reverse network toxicology, molecular simulation, and cellular perturbation experiments.
Analysis of 36,025 cells from GSE141445 identified 14,464 malignant luminal epithelial cells. Scissor identified a recurrence-associated transcriptional state enriched for adhesion, migration, angiogenesis, and proliferation programs. H&E-derived features from 304 paired TCGA-PRAD cases captured variation in this transcriptional program and supported internal recurrence risk stratification. Cross-cohort modeling prioritized CDC20, ENSA, and PTTG1. Reverse toxicology further prioritized benzo[a]pyrene (BaP), and CDC20 showed the most favorable predicted BaP docking score. In PC-3 and DU145 cells, 10 nM BaP increased CDC20 expression, whereas CDC20 silencing attenuated BaP-associated proliferation, colony formation, wound closure, and migration.
These findings identify CDC20 as a recurrence-associated molecular node involved in BaP-responsive malignant biological phenotypes and provide a phenotype-anchored framework linking recurrence biology with environmental exposure-related tumor behavior.
Xu-Chao Dai, Wei Gu, Bo Yu et al.· Frontiers in Cell and Develo...· 0 citations
Objectives Cisplatin resistance is the principal cause of relapse in high-grade serous ovarian cancer (HGSOC), but bulk-expression signatures cannot localize resistant malignant states or the tumor–microenvironment (TME) interactions that sustain them. This study aimed to define cisplatin-resistant epithelial cell states and their regulatory and metabolic circuits by integrating multi-cohort single-cell transcriptomes with pharmacogenomic and clinical data. Methods We assembled 159,419 cells from 32 HGSOC tumors across six public single-cell RNA-sequencing cohorts and performed harmonized integration, clustering, and lineage annotation. Cisplatin response was mapped to single cells by coupling scRNA-seq data to Genomics of Drug Sensitivity in Cancer predicted cisplatin response score values using Scissor, with AUCell-based validation. We then applied receptor–ligand–based cell–cell communication analysis (CellChat), transcription-factor (TF) activity inference (SCENIC and NetAct with TRRUST), pseudotime trajectory reconstruction (Monocle3), and pathway-level metabolic scoring. Associations with drug sensitivity and patient outcome were evaluated in ovarian cancer cell lines and The Cancer Genome Atlas (TCGA) HGSOC cohort. Results Among 34 epithelial subclusters, 14 were significantly enriched for a cisplatin-resistant phenotype and collectively accounted for most predicted resistant cells. These states were transcriptionally characterized by stress, interferon, and apoptotic programs and formed dense communication hubs with endothelial cells, fibroblasts, myeloid cells, and T/NK cells via extracellular-matrix and adhesion pathways (for example, LAMA3–CD44, COL6A1/2–CD44, NECTIN3–NECTIN2, and CD99–CD99 interactions). TF-activity modeling converged on an IRF1-centered regulatory program that increased along an epithelial trajectory and coordinated inflammatory and apoptotic gene expression. Metabolically, resistance-enriched epithelial states showed selective up-regulation of glycosaminoglycan biosynthesis, particularly keratan sulfate, coupled to heightened glycolysis; glycolytic activity correlated with predicted cisplatin response score in cell lines and an IRF1/STAT1 readout (GBP3) stratified survival in TCGA HGSOC. Conclusion Cisplatin resistance in HGSOC is encoded in discrete IRF1-driven epithelial states that are supported by specific TME communication networks and a glycosaminoglycan–glycolysis metabolic axis. This integrative single-cell informatics framework yields testable biomarkers and therapeutic targets for overcoming platinum resistance in ovarian cancer.
Zhaoyang Jia, Wen-Jing Pan, Xibo Zhao et al.· Cancer Informatics· 0 citations
Single-cell transcriptomics of publicly available BLCA data identifies two transcriptionally distinct fibroblast states in adjacent tissue and supports an exploratory ligand–receptor interaction framework, warranting prospective validation with primary CAF populations and adequately powered multi-specimen cohorts.
Yan-Dong He, Wen-Long Lu, Guan-Qun Ju et al.· Frontiers in Cell and Develo...· 0 citations
ABSTRACT Tongue squamous cell carcinoma, the most aggressive subtype of oral squamous cell carcinoma, which is associated with high mortality, frequent recurrence, and early lymph‐node metastasis. Triptolide, a bioactive diterpenoid from Tripterygium wilfordii, exhibits broad anticancer activity. Although its inhibitory action on OSCC has been established, its specific molecular regulatory mechanisms in TSCC, a distinct and aggressive subtype, remain largely underexplored. The present study investigates the regulatory role of triptolide in the PI3K–AKT signaling pathway, which is a critical axis intimately linked to tumor survival and metastasis. A network‐pharmacology workflow was used to intersect triptolide putative targets with TSCC‐related genes. Core hubs were identified with Cytoscape; KEGG and GO analyses highlighted the PI3K–AKT axis as the top enriched pathway. Binding kinetics were quantified by molecular docking and surface plasmon resonance (SPR). Cellular assays (CCK‐8, Transwell, wound‐healing, Western blot and immunofluorescence) validated the predictions. We retrieved 389 common targets and shortlisted eight key nodes (JUN, MAPK1, MAPK3, AKT1, TP53, BCL2, STAT3, MYC). SPR revealed high‐affinity binding of triptolide to AKT1 and PI3K. Functionally, triptolide dose‐dependently suppressed TSCC cell proliferation, invasion, and migration, downregulated PI3K–AKT signaling proteins, and reduced the colocalization and phosphorylation levels of p‐PI3K and p‐AKT. This study is the first to demonstrate the pivotal role of the PI3K–AKT signaling pathway in mediating the antitumor effects of triptolide in tongue squamous cell carcinoma. Our findings thereby provide a novel theoretical foundation for the development of traditional Chinese medicine‐based therapeutic strategies targeting this malignancy.
Shan Wang, Hong-Cheng Wei, Lin Jiang et al.· Kaohsiung Journal of Medical...· 0 citations
Colorectal cancer (CRC) exhibits marked cellular heterogeneity, and the cellular context of malignancy-associated epithelial programs remains incompletely defined. We integrated 2,993 CRC samples spanning bulk RNA-seq (n = 2,568; two OS/RFS cohorts), scRNA-seq (281,961 cells/152 specimens), spatial transcriptomics (n = 6), and proteomics (n = 267). Analyses included single-cell integration/annotation, GSVA/HALLMARK, interactome, pseudotime, and ligand–receptor mapping; functional CRISPR assays, EMT immunoblotting, and xenografts; TF profiling (SCENIC/JASPAR/ChIP-qPCR); and exploratory drug-response prediction (OncoPredict), cell-sensitivity assays, and docking/MD modeling. We constructed a stage-stratified single-cell atlas and resolved eleven malignant epithelial subsets, characterizing Epi_4 as late-stage–enriched with EMT, hypoxia, and inflammatory programs and adverse OS/RFS. GPRC5A marked this subset, which we define as GPRC5A+Epi; its expression rose from stage I→IV and was associated with poor outcomes across cohorts, with concordant spatial/proteomic observations. GPRC5A perturbation affected CRC proliferation, migration/invasion, EMT, and xenograft tumorigenicity, supporting a functionally important role in the tested models. SCENIC and ChIP-qPCR supported FOSL1 as an upstream regulator that occupies the GPRC5A promoter. Spatial and ligand–receptor analyses predicted close association and potentially reciprocal signaling between GPRC5A+Epi and POSTN+fibroblasts (COL1A1–SDC4, COL1A1/1A2–ITGA2/ITGB1, PPIA–BSG); concurrent high GPRC5A+Epi/POSTN+Fib signatures were associated with inferior OS/RFS. Drug-response analyses identified an association between GPRC5A status and trametinib sensitivity. Docking/MD produced a computational model of a possible trametinib–GPRC5A interaction, which remains experimentally unvalidated. GPRC5A⁺Epi is a malignancy-associated epithelial state in CRC, and GPRC5A is functionally important for malignant phenotypes in the tested models. Its inferred relationships with POSTN⁺ fibroblasts and the trametinib findings should be regarded as hypothesis-generating pending functional crosstalk, direct-binding, and therapeutic validation.
Wei-Chun Tang, Peng-Chen Xu, Sheng-Li Wang et al.· Journal of Translational Med...· 0 citations
SpaPred provides a hypothesis-generating framework for investigating spatial heterogeneity and candidate therapeutic vulnerabilities in HCC and prioritized Oxaliplatin, Belinostat, and Temsirolimus as candidate compounds associated with LE-related transcriptional programs.
Shi-Bo Zhang, Ziqiao Li, Kexin Yu et al.· International Journal of Mol...· 0 citations
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