Bladder cancer (BC) is aggressive and difficult to treat, with chemotherapy facing challenges like drug toxicity and resistance. Proanthocyanidins (PAC) show promising antitumor activity, as demonstrated by studies in several cancer types. However, much is still unknown about how PAC could be used as therapeutic adjuvants in BC. This study evaluated the antiproliferative potential of PAC from the black wattle (Acacia mearnsii) bark extract (BWP) in BC T24 cells using in vitro and in silico approaches. BWP treatment reduced T24 cell viability in a dose-dependent manner, with an IC50 value of 11.67 µg/mL. Flow cytometry revealed BWP-induced apoptosis in T24 cells. Additionally, nuclear morphometric analysis indicated a shift toward a senescence-like phenotype, consistent with morphological features of senescent cells. Using an in silico systems biology approach, we identified candidate genes potentially associated with BWP activity, including BACE1, DNMT1, NOS1, NOS2, NOS3, PPARA, PPARG, PPARGC1A, and PTGS2. The mRNA expression analysis (qRT-PCR) showed that BWP modulated the expression of DNMT1, BACE1, PPARG, and NOS3 in T24 cells. suggesting potential therapeutic roles in cancer progression. In conclusion, BWP exhibits antiproliferative activity and induces apoptosis in T24 bladder cancer cells, and its constituent compounds interact with candidate genes relevant to cancer biology. These findings provide a hypothesis-generating framework for future mechanistic and functional studies on PAC-based therapies.
Atypical cell fusion events can disrupt the normal post-fusion process, initiating genome chaos through the merging of parental genomes accompanied by severe and complex gross structural changes. Consequently, unscheduled fusion events can accelerate tumorigenesis by fostering persistent genomic instability, serving as catalysts in the post-hybrid selection process and decisively contributing to the acquisition of novel and malignant properties. Understanding how hybrid cells survive after fusion and identifying specific transcriptional and phenotypic signatures that characterize hybrids across different fusion phases may provide valuable insights to counteract the evolution of cancer-initiating hybrids. Two distinct populations of H2B-fluorescence-labeled M13SV1 breast epithelial cells were cocultured to yield H2B double-fluorescence-tagged hybrid clones derived from spontaneous cell fusion. Hybrid clones were picked and propagated individually to establish early- and late-passage hybrid populations. Hybrid clone and passage-dependent cell fate studies were conducted via live-cell imaging and high-resolution 3D imaging to capture aberrant mitotic outcomes. Hybrids were evaluated for ploidy status, cell cycle DNA damage, cell/nuclear morphology and transcriptomic profiling using a set of distinct methods, including flow cytometry (ploidy/DNA damage analysis), diverse cell metrics and image processing software (cell/nuclear scoring) and RNA sequencing (transcriptome profiling). Subsequently, EMT marker expression, migration/invasion, and colony- and sphere-forming capacities were investigated. Near-tetraploid hybrids undergo a hybrid clone-dependent ploidy reduction process upon long-term passaging, which is associated with cell shrinkage. Ploidy reduction was not coupled with substantial loss of DNA lesions but rather correlated with the degree of nuclear deformation. Hybrids were extremely chromosomally unstable, as evidenced by a high incidence of aberrant mitoses, nuclear pleomorphism, and elevated scores of micro- and multinucleation. Additionally, a subset of hybrids underwent ploidy reversal, characterized by endoreplication, cytokinesis failure or refusion, leading to the formation of exceptionally large or multinucleated cells. Transcriptome analysis revealed a differentially expressed gene profile of the hybrids associated with EMT, migratory aptitude and cancer-related traits, as validated by migration/invasion assays, Western Blots and colony/mammosphere assays. Collectively, the data highlight the catastrophic potential of atypical fusion processes, wherein widespread and large-scale genomic changes can occur at a rapid macroevolutionary pace and potentiate the risk for subclonal heterogeneity and malignancy.
Ubiquitin-specific protease 7 (USP7), a core member of the deubiquitinating enzyme family, acts as a double-edged sword in tumorigenesis and progression by dynamically interacting with multiple substrate proteins via its TRAF domain. Beyond tumorigenesis, USP7 is indispensable for embryonic development and genome stability, and constitutive knockout in mice leads to early embryonic lethality, indicating its essential roles in maintaining physiological homeostasis.USP7 exerts context-dependent dual effects during tumorigenesis and progression. In most p53 wild-type malignancies under basal unstressed conditions, USP7 preferentially stabilizes MDM2 to accelerate p53 turnover, which facilitates tumor growth; yet under DNA damage stress, USP7 can directly deubiquitinate and activate p53 to trigger tumor-suppressive programs. Conversely, in p53-mutant or p53-null tumors lacking functional p53 checkpoint, USP7 predominantly sustains oncogenic phenotypes by stabilizing Myc, NF-κB, EZH2 and other oncoproteins, fueling proliferation, metabolic reprogramming, immune evasion and therapy resistance.USP7 regulates a wide spectrum of biological processes, including DNA damage repair, epigenetic modification, cell cycle progression and apoptosis. Although great advances have been made in the development of USP7-targeted small-molecule inhibitors, their clinical translation is hindered by on-target toxicity, insufficient selectivity, poor pharmacokinetics and context-dependent efficacy. Future research should focus on developing highly selective inhibitors, elucidating context-specific regulatory networks, exploring predictive biomarkers and optimizing combination regimens. As a druggable deubiquitinase target, USP7 facilitates the development of novel strategies to reverse clinical chemoresistance and implement personalized oncology.
Despite advances in multimodal therapy, effective treatments for aggressive oral squamous cell carcinoma (OSCC) remain limited, underscoring the need for actionable therapeutic targets. Metadherin (MTDH), a stress-responsive gene broadly upregulated in diverse malignancies and proposed as a cancer fitness gene, has not been fully characterized in OSCC. To define its pathological and functional relevance, we integrated analyses of patient specimens, CRISPR/Cas9-mediated loss-of-function models, and transcriptomic profiling. Immunohistochemical evaluation of 100 surgically resected OSCC cases demonstrated spatial heterogeneity of MTDH expression, with enrichment at the invasive front compared with the tumor center. High MTDH expression was significantly associated with non-cohesive invasion patterns. In OSCC cell lines, MTDH knockout suppressed migration and invasion without affecting proliferation or apoptosis, effectively uncoupling invasive capacity from cell growth. Transcriptomic analysis revealed selective transcriptional modulation rather than global gene expression changes. Within this restricted gene repertoire, keratin 14 (KRT14) was consistently downregulated and positively correlated with MTDH expression at both transcript and protein levels. Collectively, these findings identify MTDH as a determinant of OSCC invasion and migration, with KRT14 as a candidate downstream effector, without broadly perturbing global transcription, supporting MTDH as a precision therapeutic target for inhibiting invasion.
