This study aimed to investigate the expression, biological functions, and underlying mechanisms of HNRNPC in clear cell renal cell carcinoma (ccRCC). The expression of HNRNPC in ccRCC tissues and cell lines was detected. The effects of HNRNPC knockdown and overexpression on ccRCC cell proliferation, migration, and invasion were analyzed using cellular models. The role of HNRNPC in recognizing m6A modifications and regulating AURKB mRNA stability was validated through treatment with the methylation inhibitor STM-245, an actinomycin D stability assay, a dual-luciferase reporter assay, and RNA immunoprecipitation. A nude mouse xenograft model was established for in vivo functional validation. The results showed that HNRNPC was significantly overexpressed in ccRCC tissues and cell lines. Knockdown of HNRNPC suppressed cell proliferation, sphere formation, migration, and invasion, while overexpression promoted these malignant phenotypes. Mechanistically, HNRNPC enhanced the stability and expression of AURKB mRNA by recognizing and binding to m6A modification sites on AURKB mRNA. HNRNPC overexpression reversed the suppression of cellular phenotypes induced by AURKB knockdown. In vivo experiments demonstrated that knockdown of either HNRNPC or AURKB significantly inhibited tumor growth and downregulated Ki-67 expression. In conclusion, HNRNPC contributes malignant progression of ccRCC, at least in part, by recognizing m6A modifications on AURKB mRNA and enhancing its stability, suggesting that the HNRNPC/AURKB axis may serve as a potential therapeutic target for ccRCC.
Cancer immunophenotypes—immune desert (ID), immune excluded (IE), and immune inflamed (II)—reflect intratumoral immune activity and correlate with immunotherapy efficacy. Serological methods for non-invasive immunophenotyping would be clinically valuable for selecting therapies. We measured plasma concentrations of cytokines (IL-6, IL-10, IL-16, IL-18, VEGF), chemokines (CXCL9, CXCL13), and soluble membrane molecules (sPD-1, sPD-L1, sCTLA-4, sCD163) in 49 patients with head and neck squamous cell carcinoma (HNSCC) and 22 healthy donors using an automated chemiluminescence enzyme immunoassay system. Plasma levels of IL-6, IL-10, VEGF, CXCL9, CXCL13, and sCD163 were significantly higher in patients with HNSCC than in healthy donors. IL-10 levels trended higher in the order ID > IE > II, whereas CXCL9, CXCL13, and sCD163 levels trended higher in the order II > IE > ID. This progressive trend across phenotypes was further enhanced by calculating the ratios of these markers to IL-10. Significantly higher sPD-1 levels were observed in the II phenotype. These results indicate that plasma concentrations and ratios of CXCL9, CXCL13, sCD163, IL-10, and sPD-1 serve as potential surrogate biomarkers reflecting tumor immunophenotypes in HNSCC.
Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma, characterized by dysregulated lipid metabolism and therapy resistance, leading to a poorer prognosis than other subtypes. This study investigates the role of the immune checkpoint molecule CD276 in lipid metabolic reprogramming and the regulation of ferroptosis in ccRCC. Clinical sample analysis, in vitro experiments, and animal models revealed that CD276 is significantly overexpressed in ccRCC and positively correlated with an unfavorable prognosis. Mechanistically, CD276 activates the transcription factor sterol regulatory element-binding protein 1 (SREBP1), upregulates the expression of fatty acid synthase (FASN), promotes de novo fatty acid synthesis, and drives lipid accumulation. Concurrently, CD276-mediated lipid metabolic reprogramming suppresses ferroptosis in ccRCC cells by increasing reduced glutathione levels and enhancing glutathione peroxidase 4 activity. In vivo experiments confirmed that inhibiting CD276 significantly suppresses tumor growth and enhances the efficacy of ferroptosis inducers. This study reveals the pivotal role of the CD276–SREBP1–FASN axis in regulating lipid metabolism and ferroptosis in ccRCC, providing a theoretical basis for CD276-targeted therapy combined with ferroptosis induction as a treatment strategy for ccRCC.
Spindle cell/sclerosing rhabdomyosarcoma (SRMS) comprises a heterogeneous group of tumors characterized by distinct molecular alterations and clinical behaviors. Among them, ZFP64::NCOA3 fusion-positive SRMS represents an extremely rare subtype, and its biological characteristics remain poorly understood because of the lack of representative experimental models. In this study, we established a patient-derived organoid (PDO) line, designated OICI-RMS-1275, together with organoid-derived xenograft (ODX) models from a previously reported case of ZFP64::NCOA3 fusion-positive SRMS using a modified air–liquid interface organoid culture method. The established organoids exhibited stable long-term propagation and retained tumorigenic capacity following serial transplantation into NOD-scid IL2Rgnull mice. Histological and immunohistochemical analyses demonstrated preservation of the characteristic spindle cell morphology and diffuse expression of MyoD1 and PAX7 observed in the original tumor. Polymerase chain reaction and Sanger sequencing confirmed retention of the ZFP64::NCOA3 fusion transcript and fusion breakpoint in the organoids and ODXs. Lentiviral-mediated knockdown of the fusion gene significantly suppressed organoid proliferation, indicating dependency on ZFP64::NCOA3 for tumor growth. Transcriptomic analyses revealed preservation of major transcriptional features between the original tumor and ODXs, including myogenic lineage-associated gene expression programs. Fusion-gene suppression decreased expression of MYOD1 and DLK1 while increasing PPARG expression, suggesting involvement of the fusion in lineage-associated transcriptional regulation. Under adipogenic induction conditions, SRMS organoids exhibited lipid accumulation detected by Oil Red O staining regardless of fusion-gene suppression status, indicating preserved adipogenic differentiation potential. These findings establish the first PDO and ODX models of ZFP64::NCOA3 fusion-positive SRMS and demonstrate fusion-gene dependency in this rare molecular subtype. Our results further suggest that despite strong myogenic transcriptional programs, fusion-positive SRMS retains adipogenic differentiation capacity. These models provide a valuable platform for investigating disease biology and developing novel therapeutic strategies for rare fusion-positive rhabdomyosarcomas.
The role of SPIN.DOC in tumorigenesis remains unclear. In this study, we utilized single-cell RNA sequencing (scRNA-seq) data to investigate SPIN.DOC expression in a cohort of normal, primary human colorectal cancer (CRC) and metastatic tissues from CRC patients. Our findings revealed differential expressions of SPIN.DOC across multiple cell clusters in different conditions and with the highest expression observed in tumor samples, suggesting its potential role in CRC progression. Further analysis showed that SPIN.DOC positive cell-cluster showed upregulation of β-catenin and cancer stem cell (CSC) marker genes, indicating its role in activating Wnt signaling for cell proliferation. Additionally, these cells also showed downregulation of genes associated with cell–cell junctions, promoting epithelial-mesenchymal transition (EMT) and cancer progression in colon tumors. To validate these findings experimentally, we analyzed the expression of SPIN.DOC in normal colorectal and three colorectal cancer cell lines by immunofluorescence and western blotting. SPIN.DOC was expressed at low level in human normal colon epithelial cells but was highly expressed in all the three colorectal cancer cell lines tested. Consistent with the scRNA data, overexpression of SPIN.DOC in CRC cell line, enhanced cell proliferation, cell migration, invasion, colony-forming capability and Wnt signaling. Moreover, these cells upregulate EMT-associated genes and cancer stem cell markers, as determined by qRT-PCR and immunoblotting. Enhanced spheroid formation in SPIN.DOC overexpressing cells further indicated increased stemness of the cells. Conversely, the knockdown of SPIN.DOC diminished the aforesaid phenotypes. Overall, these data suggest that SPIN.DOC promotes cancer cell proliferation, metastasis and regulates pluripotency and self-renewal of colorectal CSCs. Overall, our study highlights SPIN.DOC as a key essence of CRC by regulating cancer stemness and provides an opportunity of using SPIN.DOC, as a diagnostic and prognostic biomarker of colorectal cancer.
Carcinoma of unknown primary (CUP) remains a clinically challenging malignancy with limited treatment options and poor prognosis. Although genomic profiling has identified potentially actionable alterations, optimal therapeutic strategies are often unclear. Here, we report the establishment and characterization of a patient-derived tumoroid from an EGFR-amplified CUP and evaluate its utility as a functional model for therapeutic assessment. The patient-derived tumoroid (PDT-CUP#1) was successfully established from resected tumor tissue and maintained in long-term culture. Whole-exome sequencing demonstrated genomic concordance between the PDT-CUP#1 and the original tumor. Quantitative PCR and fluorescence in situ hybridization confirmed EGFR amplification. Xenograft tumors derived from PDT-CUP#1 recapitulated the histopathological features of the original lesion. Functional drug testing demonstrated differential sensitivity among EGFR-targeted agents. Whereas EGFR tyrosine kinase inhibition resulted in modest growth suppression, anti-EGFR monoclonal antibodies exhibited moderate antitumor effects, and an EGFR-targeted antibody–drug conjugate showed the most pronounced growth inhibition. Notably, the therapeutic sensitivity observed in the tumoroids was generally consistent with the patient’s clinical response to platinum-based chemotherapy combined with necitumumab. These findings demonstrate that a CUP-derived tumoroid can be successfully established while preserving the key molecular and pathological features of the original tumor. This model provides a platform for functional evaluation of therapeutic vulnerabilities and offers proof-of-concept for integrating patient-derived tumoroids into therapeutic decision-making in CUP.
Chemoresistance remains a major obstacle to improving outcomes in colorectal cancer (CRC), particularly among patients receiving 5-fluorouracil (5-FU)-based therapy. Here, we developed a READ-derived response-associated gene signature for recurrence stratification and performed exploratory cross-cohort evaluation in additional colorectal cancer cohorts, while further exploring its association with treatment-response phenotypes. In the GSE190826 cohort, pathological complete response (pCR) versus non-pCR status was used as a clinically annotated treatment-response phenotype for downstream screening. Using differential expression analysis in TCGA-READ, we identified 4277 differentially expressed genes and constructed a weighted gene co-expression network analysis (WGCNA) to define clinically relevant modules. Genes from READ-related modules were further screened by logistic regression in GSE190826 and univariate Cox regression in TCGA-READ, yielding six prognostic genes associated with pathological non-response. Least absolute shrinkage and selection operator (LASSO)–Cox regression subsequently refined these candidates to a four-gene signature, including ENHO, BLACAT1, LEMD1, and ASNS, which was used to establish a READ-derived response-associated gene (RDRG) risk score. High-risk patients showed poorer recurrence-free survival in the TCGA-READ training set and an exploratory TCGA-READ testing subset, and exploratory evaluations in additional colorectal cancer cohorts (TCGA-COAD and GSE17536) showed similar recurrence-stratification trends, with moderate time-dependent AUC values in TCGA-READ (3-year AUC = 0.766; 5-year AUC = 0.714). Multivariate Cox analyses showed that the risk score remained associated with RFS after adjustment for clinical covariates, and a nomogram incorporating disease stage provided additional exploratory prognostic stratification. The RDRG score also showed only modest and exploratory associations with treatment-response phenotypes across multiple datasets. Single-cell RNA-seq analysis revealed that malignant cells contributed the highest risk scores, with enrichment of epithelial-mesenchymal transition (EMT), KRAS signaling, coagulation, and hypoxia-related programs in high-risk tumor cells. Among the four genes, LEMD1 showed the strongest malignant-cell enrichment and was further examined in functional assays, where LEMD1 knockdown reduced CRC cell migration and increased 5-FU sensitivity, supporting its role as a candidate functional contributor rather than a validated mechanistic driver. Collectively, the RDRG risk score may serve as a READ-derived model for recurrence stratification, with exploratory cross-cohort applicability in broader colorectal cancer cohorts and exploratory associations with treatment-response phenotypes. The same risk-score formula was applied across cohorts, whereas high- and low-risk thresholds were cohort-specific rather than fixed from the training cohort.
Ovarian endometriosis is a chronic inflammatory disease characterized by extensive tissue remodeling and fibrosis, which significantly impacts female reproductive health. This study demonstrates that ectopic endometrium (Ect) in patients with endometriosis exhibit pronounced fibrotic changes compared to normal (Norm) and eutopic endometrium (Eut), characterized by the upregulation of fibrosis-associated genes including α-SMA, Col-1α1, and CTGF. Using primary cell isolation and characterization, we identified that exosomes derived from ectopic endometrial stromal cells (EctESCs-exo) serve as critical mediators of this pathological process. Fluorescent tracking and functional assays revealed that EctESCs-exo are internalized by eutopic endometrial stromal cells (EutESCs), subsequently triggering a pro-fibrotic phenotype in vitro and promoting lesion growth and collagen deposition in a mouse model of endometriosis. Through bioinformatic and multi-cohort validation, miR-21-5p was found to be significantly enriched in both ectopic tissues and their secreted exosomes. Silencing miR-21-5p in donor ectopic endometrial stromal cells (EctESCs) effectively attenuated the fibrotic response in recipient cells, whereas miR-21-5p-enriched exosomes markedly accelerated disease progression and fibrosis in vivo. Mechanistically, dual-luciferase reporter assays and functional rescue experiments confirmed that miR-21-5p directly targets von Hippel-Lindau (VHL), a negative regulator of fibrosis that is downregulated in ectopic lesions. Collectively, our findings elucidate a novel mechanism of exosome-mediated intercellular communication where EctESCs drive fibrotic progression via the miR-21-5p/VHL axis, suggesting that targeting this pathway could offer a promising therapeutic strategy for endometriosis.
Uterine corpus endometrial carcinoma (UCEC) ranks as the 6th most common malignancy among women. Emerging evidence indicates that the dysregulation of tRNA-derived fragments (tRFs) is involved in the pathogenesis of multiple cancer types, including UCEC; however, the molecular mechanisms underlying the roles of tRFs in UCEC remain poorly characterized. Desmocollin3 (DSC3), a transmembrane protein, is predominantly expressed in the basal and suprabasal layers of normal stratified epithelia. While accumulating evidence has implicated DSC3 in the pathogenesis of multiple disease entities, its functional role in UCEC remains elusive. The present study is designed to investigate the functional significance and underlying molecular mechanisms of tRF-19-79MP9PJZ in the progression of UCEC. In this study, tRF-19-79MP9PJZ was found to be significantly upregulated in UCEC tissues and cell lines, with its elevated expression correlating with unfavorable prognostic outcomes in UCEC patients. Furthermore, tRF-19-79MP9PJZ knockdown was observed to suppress the proliferative and migratory capacities of UCEC cells, while concurrently enhancing apoptotic processes. At the mechanistic level, tRF-19-79MP9PJZ was demonstrated to facilitate UCEC progression through targeted regulation of DSC3. Collectively, this study elucidates a previously uncharacterized mechanism whereby tRF-19-79MP9PJZ drives UCEC development, thereby highlighting the potential of the tRF-19-79MP9PJZ/DSC3 axis as a therapeutic target for UCEC intervention.
Regular physical exercise can induce a multifaceted cardioprotective phenotype characterized by improved Ca2⁺ handling, mitochondrial resilience, redox buffering, autonomic regulation, and resistance to ischemia–reperfusion injury. Ca2⁺/calmodulin-dependent protein kinase II (CaMKII), particularly cardiac CaMKIIδ, is positioned at the intersection of these adaptive and maladaptive responses because it couples repetitive Ca2⁺ oscillations to excitation–contraction coupling, ion-channel regulation, transcriptional remodeling, mitochondrial stress signaling, and cell-death pathways. Current evidence indicates that CaMKII is not intrinsically protective or harmful; rather, its biological output depends on activation magnitude, duration, post-translational modification, isoform or splice-variant composition, and subcellular localization. Within physiological exercise contexts, transient and compartmentalized CaMKII signaling may support rate adaptation, phospholamban phosphorylation, sarcoplasmic reticulum Ca2⁺ reuptake, and contractile reserve. In contrast, chronic oxidative, inflammatory, catecholaminergic, or metabolic stress promotes autonomous CaMKII activation, RyR2-mediated Ca2⁺ leak, late Na⁺ current, mitochondrial dysfunction, arrhythmogenesis, and adverse remodeling. Exercise training appears to normalize this pathological signaling environment by improving redox and metabolic homeostasis, mitochondrial quality control, nitric oxide bioavailability, and autonomic balance, while preserving physiological CaMKII-dependent cardiac reserve. In this review, we synthesize current evidence on CaMKII as a context-dependent mediator of exercise-induced cardioprotection and discuss its implications for cardiovascular disease mechanisms, biomarker development, exercise prescription, and selective CaMKII-targeted therapy.
Platelets are major circulating reservoirs of transforming growth factor-β1 (TGF-β1). After tumor-associated activation, they release latent TGF-β1 from α-granules or present it at the platelet surface through glycoprotein A repetitions predominant. The latent complex is then activated by integrins, proteases, mechanical forces, and local physicochemical conditions. Direct platelet-specific evidence supports roles for platelet-derived TGF-β1 in established tumor growth, epithelial–mesenchymal transition, immune suppression, and metastasis. Platelet-associated studies also suggest effects on stromal remodeling, circulating tumor cell adaptation, endothelial dysfunction, and pre-metastatic niche formation. However, the platelet origin of TGF-β1 remains unresolved in many settings. This review distinguishes platelet-specific evidence from platelet-associated findings and source-independent mechanistic extrapolations. It also examines whether the stage-dependent paradox of TGF-β signaling applies to platelet-derived TGF-β1. The current evidence mainly supports a tumor-promoting role in established and metastatic disease, whereas its role in tumor initiation remains unclear. We further compare systemic therapies with platelet-directed, platelet-guided, and other targeted TGF-β interventions.
Germline variants in leukemia predisposition genes are increasingly detected in pediatric patients. Nevertheless, the rare variants identified in diagnostic samples may be misinterpreted without variant-level functional evaluation and cautious clinical interpretation. Whole-exome sequencing was performed on 28 individuals from five kindreds in which at least two children developed acute leukemia. Results identified a rare ETV6 variant, c.604C > G (p.Arg202Gly), in monozygotic twins with ETV6::RUNX1-positive B-cell precursor acute lymphoblastic leukemia. To support variant interpretation, HeLa-cell populations stably expressing FLAG-tagged wild-type ETV6 and p.Arg202Gly and the known loss-of-function control p.Pro214Leu were established. Subcellular localization was assessed via immunofluorescence microscopy with quantitative scoring. Transcriptional repression was evaluated using luciferase reporter assays driven by ETV6 target promoters (MMP3 and PF4). p.Arg202Gly predominantly showed nuclear localization comparable to WT, whereas p.Pro214Leu was enriched in the cytoplasm. In the reporter assays, similar to WT, p.Arg202Gly retained repression activity. Meanwhile, p.Pro214Leu failed to repress both reporters. The in-silico prediction of nuclear export sequences suggested an additional export signal in p.Pro214Leu, but not in p.Arg202Gly. Collectively, these findings indicate that p.Arg202Gly behaves as WT-like in the assays performed and do not support a loss-of-function effect. Our study emphasizes the importance of variant-level functional assessment for rare ETV6 variants to inform clinical interpretation and avoid overestimation of pathogenicity.
Cancer cachexia is a debilitating systemic syndrome that affects a substantial proportion of patients with advanced malignancy and is associated with impaired treatment tolerance, reduced quality of life, and increased mortality. While skeletal muscle wasting is a defining clinical feature, cachexia involves coordinated dysfunction across multiple organs, yet it remains unclear whether cachexia imposes a unified, body-wide transcriptional program or primarily induces organ-specific responses. Here, we leveraged an isogenic xenograft model derived from human duodenal neuroendocrine carcinoma in which the cachexia-inducing subline AkuNEC was established through in vivo serial passaging from the parental, largely non-cachexia-inducing line TCC–NECT-2. We performed bulk RNA sequencing of skeletal muscle, liver, kidney, and heart from cachectic AkuNEC-bearing mice, non-cachectic TCC–NECT-2-bearing mice, and uninoculated controls. Differential expression analyses identified organ-dependent sets of transcripts associated with cachexia. However, unsupervised analyses of global expression patterns consistently showed that tissue identity dominated transcriptome structure and samples did not segregate by cachexia status. In addition, comparisons of tumor-bearing vs uninoculated controls revealed broadly similar transcriptional shifts for AkuNEC and TCC–NECT-2 within each organ. Together, these data indicate that cachexia-associated transcriptional changes are present but remain modest relative to dominant tissue-specific programs at the whole-transcriptome level. The AkuNEC/TCC–NECT-2 system provides a controlled platform for future studies incorporating cell-type-resolved, spatial, and multi-omic approaches to delineate the mechanisms linking tumor evolution to multi-organ remodeling in cancer cachexia.
Acute myeloid leukemia (AML) remains a highly lethal hematologic malignancy characterized by metabolic reprogramming, therapeutic resistance, and poor survival, particularly in older patients. Nicotinamide adenine dinucleotide (NAD⁺) metabolism has emerged as a central driver of AML progression, and recent studies have identified solute carrier family 25 member 51 (SLC25A51) as the primary mitochondrial NAD⁺ transporter in mammalian cells. SLC25A51 regulates mitochondrial redox balance, oxidative phosphorylation, and tricarboxylic acid (TCA) cycle activity, thereby sustaining leukemic proliferation and survival. Structural studies have elucidated its six-transmembrane helix architecture, salt-bridge-mediated transport mechanism, and stabilization by cardiolipin binding. Functional investigations demonstrate that SLC25A51 overexpression correlates with poor prognosis, while its depletion disrupts mitochondrial metabolism, induces apoptosis, and suppresses AML progression in vivo. Therapeutically, pharmacologic inhibition of SLC25A51 with fludarabine, or its combination with hypomethylating agents, such as 5-azacytidine, enhances antileukemic efficacy by perturbing metabolic and epigenetic regulation. Moreover, SLC25A51 expression may serve as a predictive biomarker for mitochondrial-targeted therapies, such as complex I inhibitors. Future translational research should focus on developing selective inhibitors, optimizing combination strategies with demethylating agents and BCL-2 inhibitors, and validating its prognostic significance in clinical cohorts. Collectively, SLC25A51 represents a promising metabolic target with potential to overcome therapeutic resistance and improve patient outcomes in AML. Furthermore, this review discusses its potential implications across distinct genetic subtypes of AML (e.g., mutations in TP53, NPM1, and RAS), thereby highlighting key directions for future translational research.
Hepatocellular carcinoma (HCC) frequently develops resistance to lenvatinib, a multikinase inhibitor, necessitating the development of novel therapeutic strategies. Here, we identify bikinin as a potent eIF5A2 inhibitor through structure-based virtual screening (> 100,000 compounds) and demonstrate its synergistic effect with lenvatinib in HCC cells. Mechanistically, bikinin suppresses deoxyhypusine synthase (DHS)-mediated hypusination of eIF5A2, thereby downregulating the expression of transcription factor EB (TFEB). Furthermore, while DHS knockdown enhanced the sensitivity of HCC cells to lenvatinib, the addition of bikinin treatment provided no further significant sensitization. We also observed that the combination of bikinin and lenvatinib significantly promoted apoptosis and suppressed proliferation in HCC cells. Although TFEB overexpression conferred resistance to lenvatinib and activated autophagy, these effects were reversed by co-treatment with bikinin, which restored lenvatinib sensitivity and inhibited autophagic flux. Bikinin disrupts TFEB-driven autophagy, as evidenced by reduced LC3-II conversion, p62 accumulation, and decreased autophagosome formation. In in vivo experiments, the combination therapy with lenvatinib and bikinin achieved marked tumor regression, accompanied by suppressed Ki-67 expression and elevated TUNEL positivity. Finally, RNA-seq data identified TFEB downregulation as a critical mediator of this therapeutic sensitization. Our work unveils a novel therapeutic axis wherein targeting eIF5A2 hypusination disrupts TFEB-dependent autophagy to overcome lenvatinib resistance in HCC cells.
Human tumor-derived cell lines remain foundational and complementary resources for mechanistic and translational cancer research. However, reports of newly established cell lines vary in the scope and completeness of basic information, and the distinction between essential reporting items and value-added characterization is often unclear. This review proposes a pragmatic Minimum reporting checklist for newly established human tumor-derived cell lines. The Minimum tier prioritizes durable, reproducibility-critical information required for establishment reports. These items include unambiguous cell line designation and Research Resource Identifier (RRID) citation, identity authentication through short tandem repeat (STR) profiling, mycoplasma status, tumor and donor provenance, ethics oversight, reproducible culture conditions, passage history and/or population doubling level, and a transparent access pathway. Tumor and donor provenance includes the diagnostic basis, key tumor specimen characteristics, and limited, anonymized clinicopathological context. The checklist is anchored in the convergence of four widely used frameworks: the International Cell Line Authentication Committee (ICLAC) Cell Line Checklist, Cellosaurus/RRID, Nature Portfolio reporting standards, and UK Co-ordinating Committee on Cancer Research (UKCCCR) guidelines. It is provided as a user-facing checklist (Table 1), a mapping to ICLAC and Cellosaurus practices (Table 2), and a crosswalk showing areas of convergence across all frameworks (Online Resource 1; Table S1). Xenografts, extensive functional phenotyping, and broad omics profiling are discussed as Recommended or Optional modules rather than Minimum requirements. Adoption of this Minimum checklist may harmonize reporting practices, improve transparency and reusability, and increase the likelihood that new cell lines, including those from rare cancers, are publicly reported, registered, and discoverable.
Renal cell carcinoma (RCC) is a common and highly lethal malignancy. Tribbles pseudokinase 3 (TRIB3) has been reported to exert oncogenic roles in the pathogenesis of multiple tumors; nevertheless, the detailed mechanisms through which TRIB3 drives RCC development remain largely undefined. In this study, qPCR and Western blot were used to quantify the levels of TRIB3, STIP1 homology and U-box containing protein 1 (STUB1), and activating transcription factor 2 (ATF2). Cell proliferation, invasion, stemness, and angiogenic capacity were assessed by MTT assay, flow cytometry, Transwell invasion assay, sphere-formation assay, and tube-formation assay, respectively. The interaction between TRIB3 and STUB1 was examined by co-immunoprecipitation (Co-IP), and the relationship between TRIB3 and ATF2 was verified using dual-luciferase reporter assays. A xenograft model was employed to evaluate the tumorigenic potential of RCC cells upon STUB1 overexpression. The study found that TRIB3 expression was markedly elevated in RCC. Knockdown of TRIB3 profoundly inhibited cell proliferation, invasion, migration, and stem-like properties, while simultaneously inducing apoptosis and promoting ferroptosis. TRIB3 physically interacted with STUB1 and was poly-ubiquitinated and degraded by this E3 ligase; consequently, STUB1 overexpression suppressed RCC progression both in vitro and in vivo by targeting TRIB3. Conversely, ATF2 transcriptionally activated TRIB3 expression, and enforced TRIB3 expression partially rescued the inhibitory effects of ATF2 knockdown in RCC cells. Collectively, these findings demonstrate that TRIB3 drives RCC progression; STUB1 curbs RCC by degrading TRIB3, whereas ATF2 promotes RCC via transcriptional activation of TRIB3. Thus, modulating TRIB3 expression may offer a novel therapeutic strategy for RCC.
Claudin-7 is a basolaterally enriched tight junction protein that participates in membrane complexes with EpCAM and Trop-2, linking epithelial adhesion to barrier organization. Although proteolytic processing of EpCAM/Trop-2 has been proposed to regulate this complex, its functional consequences for claudin-7 remain unresolved. Here, we define the role of matriptase in regulating claudin-7 dynamics in HaCaT human keratinocytes. Under steady-state conditions, matriptase knockout had only modest effects on claudin-7 distribution, suggesting a limited role in initial complex formation. In contrast, induction of matriptase autoactivation by mildly acidic stress triggered rapid and pronounced remodeling of claudin-7 complex in parental cells. Claudin-7 underwent progressive redistribution from continuous junctional staining to discontinuous bar-like structures, followed by the appearance of abundant intracellular puncta consistent with endocytosed vesicles, ultimately leading to its removal from the cell surface. This dynamic process was temporally coordinated and accompanied by increased accessibility of the claudin-7 C-terminus, indicating disassembly of claudin-7 complexes. These changes were abolished in matriptase-deficient cells, in which claudin-7 remained stably associated with cell-cell interfaces and failed to internalize. Suppression of matriptase autoactivation by NaCl similarly prevented claudin-7 internalization, demonstrating that proteolytic activity is required for this process. Notably, adherens junctions remained largely intact with E-cadherin continuously localized at cell-cell interfaces. Collectively, these findings establish that matriptase proteolytic activity is essential for disassembly and internalization of claudin-7 complex. This work supports a model in which the EpCAM/Trop-2-claudin-7 axis functions as a protease-regulated membrane complex that dynamically modulates epithelial organization.
Tumor vasculature has traditionally been viewed as structurally and functionally abnormal and, therefore, is primarily targeted for inhibition. However, emerging evidence from vascular biology, regenerative medicine, and bioengineering challenges this paradigm, demonstrating that blood vessels actively instruct and shape the tumor microenvironment. Here, we propose that tumor vasculature functions as a dynamic and programmable interface that regulates cancer progression and define vascular reprogramming as a therapeutic strategy to actively redesign vascular structure, function, and signaling to control the tumor ecosystem. By integrating recent advances in endothelial cell heterogeneity, vascular niche biology, and multiscale modeling, we illustrate how tumor vessels govern cancer stemness, immune-cell trafficking, and metabolic adaptation, positioning the vasculature as a central regulatory hub rather than a passive conduit. We further highlight enabling technologies, including vascularized organoids, organ-on-a-chip systems, and iPSC-derived vasculature, that enable the precise reconstruction and manipulation of human vascular microenvironments, providing unprecedented opportunities to experimentally control vascular dynamics. Importantly, we distinguish vascular reprogramming from conventional anti-angiogenic and normalization strategies, emphasizing its potential to achieve sustained and integrative control of the tumor microenvironment. By modulating vascular permeability, perfusion, and immunoregulatory signaling, this approach enhances drug delivery, improves immune infiltration, and increases therapeutic sensitivity. Finally, we discuss the key challenges for clinical translation, including safety, scalability, and model limitations, and highlight future directions driven by spatial omics and artificial intelligence. Collectively, this framework establishes tumor vasculature as a designable therapeutic interface and advances a new paradigm in cancer therapy: not merely targeting the tumor microenvironment but engineering it through vascular control.
Intrahepatic cholangiocarcinoma (ICC) is an insidious and aggressive malignancy with poor prognosis. Our previous research has suggested that miR-7-5p modulates the ICC cell phenotype by targeting MyD88; however, its downstream molecular mechanisms remain poorly elucidated. Considering that TGF-β signaling and aerobic glycolysis provide a favorable growth environment for tumors, this study aims to explored the relationship between the miR-7-5p/MyD88 axis and these metabolic characteristics. Bioinformatics methods were used to analyze the MyD88 expression in the GSE107943, and inferred its association with TGF-β signaling activity and glycolysis scores. Next, a series of experiments was conducted to evaluate the biological functions of MyD88 and miR-7-5p, with a TGF-β activator applied to elucidate potential mechanisms. A subcutaneous xenograft mouse model was used for in vivo validation. MyD88 expression was highly expressed in ICC samples, and its levels were positively correlated with TGF-β signaling activity and glycolysis scores. MyD88 knockdown attenuated the viability, migration, and glycolysis of ICC cells, thereby inhibiting tumor growth in vivo. Furthermore, MyD88 acted in a TGF-β-dependent manner, and TGF-β activation reversed the effects of MyD88 knockdown on malignant phenotype. Experiments also showed that MyD88 downregulation was caused by miR-7-5p, and then MyD88 overexpression reversed the suppressive effect of miR-7-5p on glycolysis. Collectively, miR-7-5p specifically targets MyD88, weakening glycolysis in ICCs by reducing TGF-β signaling activity, thus exerting an inhibitory effect on ICC.
