Pembrolizumab is a novel humanized anti-PD-1 monoclonal antibody capable of enhancing T-cell mediated antitumor immunity. However, the function of pembrolizumab on tumor cells themselves and relative molecular mechanism in ovarian cancer remain unknown. Our study demonstrated pembrolizumab exerted remarkable suppressive impacts on proliferation, colony formation and migration of ovarian cancer cells in vitro. Furthermore, pembrolizumab treatment delayed cell cycle progress from G1 to S phase transition and suppressed cell growth in ovarian cancer cells. Mechanistically, pembrolizumab decreased the stability of CDK6 protein through a polyubiquitin-mediated proteasomal degradation pathway. Meanwhile, pembrolizumab treatment dose-dependently reduced Snail, Vimentin and N-cadherin expressions and enhanced E-cadherin expressions. Additionally, the combined treatment of pembrolizumab and cisplatin effectively enhanced anti-proliferative effect of cisplatin on HO-8910 cells. These findings suggested pembrolizumab efficiently suppressed malignant progression of ovarian cancer cells and facilitated proteasomal degradation of CDK6 and increased cisplatin inhibition of HO-8910 cells proliferation, therefore providing a promising therapeutic strategy for ovarian cancer.

We investigated trends in the use of perioperative therapy and the efficacy of adjuvant immunotherapy on the prognosis of patients with muscle-invasive urothelial carcinoma (MIUC). The usage and trends in neoadjuvant and adjuvant therapy were examined, and the efficacy of adjuvant immunotherapy was assessed using propensity score-adjusted Cox multivariate analysis. We investigated 1383 patients with muscle-invasive bladder cancer and 1124 patients with upper tract urothelial carcinoma; 1095 (43.7%) patients received neoadjuvant therapy and 366 (14.6%) patients received adjuvant therapy. Adjuvant therapy usage rate increased from 30.3% before 2022 to 61% after 2022 in patients with pathological high-risk cancer (pT3-4, ypT2-4, or pN+). The adjuvant immunotherapy usage rate increased from 2.8% before 2022 to 67.5% after 2022. Sixty-three (18.9%) of the 334 patients with pathological high-risk cancer who were treated with adjuvant therapy were treated with adjuvant immunotherapy. The propensity score-adjusted Cox multivariate analysis showed that adjuvant immunotherapy significantly improved disease-free survival (Hazard ratios (HR) 0.39, P < 0.005) and overall survival (HR 0.20, P < 0.005) compared with conventional adjuvant chemotherapy. In conclusion, the introduction of adjuvant immunotherapy led to the increased use of adjuvant therapy and improved prognoses in patients with MIUC in real-world practice.

Head and neck squamous cell carcinoma (HNSCC) poses a global health challenge. The management of HNSCC is complicated by the difficulty in detecting occult lymph node metastases, leading to dilemmas in elective neck dissection decisions, which will impair patients' quality of life without improving survival for nodal negative patients. We conducted a comparative analysis of the clinical features, genomic alterations, gene expression and methylation, tumor microenvironment and cellular states between the clinically N0 and pathologically N0 (cN0-pN0) patients and occult lymph node metastatic patients. Patients with occult lymph node metastases typically present with more poorly differentiated primary tumors and higher rates of angiolymphatic and perineural invasion. We identified a distinctive genomic mutation spectrum in the primary tumors of patients with occult metastases, notably in genes such as NSD1, ARHGAP15 and SMARCA4. A whole-genome DNA hypomethylation and altered gene expression profiles are identified in occult lymph node metastatic patients. Analysis of the tumor microenvironment revealed an enrichment of CARNS1 + NK cells and CBX1 + tumor cells in occult metastatic patients. In conclusion, patients with occult lymph node metastases exhibit distinct molecular and clinical features compared with cN0-pN0 patients.

Tumor associated neutrophils (TANs) exert dual and opposing functions in tumors, acting pro-tumorigenic and anti-tumorigenic, depending on tumor progression, polarization state and subtype. Consequently, the prognostic impact of TANs in breast cancer is also contradictory. Since neutrophils are critically needed to fight infections in cancer patients, the mediators leading to tumor progression need more investigation as potential future targets. The neutrophil derived mediator myeloperoxidase (MPO) is a peroxidase with dual functions in tumors, acting both immune enhancing and suppressing. Patients with metastatic breast cancer (MBC) have aggressive tumors with a dismal prognosis and urgently need novel treatment strategies. Therefore, we here aimed to investigate the prognostic impact of TANs, MPO+ TANs and MPO+ non-neutrophils using a cohort with newly diagnosed MBC patients specifically. We show that high infiltration of MPO+ TANs and MPO+ non-neutrophils in the primary tumor (PT), was associated with clinicopathological features and worse prognosis in patients with MBC. However, only infiltration of MPO+ TANs showed independent prognostic impact in multivariable analysis adjusting for other prognostic factors in MBC. The results need to be validated in a larger cohort but suggests that MPO targeting strategies could be relevant in breast cancer patients with aggressive disease.

This study investigates the concentration-dependent cytotoxicity of aluminum-doped zinc oxide nanoparticles on breast cancer (MDA-MB-231) and normal mammary epithelial cells (MCF-10 A). Pure and Al-doped ZnO nanoparticles (Al-ZnO NPs, Zn1-xAlxO, x = 0.0: ZnO, 0.01: ZA1, 0.03: ZA3, and 0.05: ZA5) were synthesized by the gelatin-based sol-gel method. The properties of the pure and Al-doped ZnO nanoparticles were investigated by X-ray diffraction (XRD), field emission electron microscopy (FESEM), and ultra-violate-visible (UV-vis) spectroscopy. Using an indirect viability assay, the prepared samples were evaluated across a 5-500 µg/mL range, with IC50 (half-maximal inhibitory concentration) as the primary metric. Results demonstrated enhanced toxicity toward cancer cells, with IC50 values for ZnO, ZA1, ZA3, and ZA5 at (225, 100, 80, and 60 µg/mL) compared to normal cells (500 µg/mL, in the experimental range), respectively. Progressive doping (ZA1 → ZA5) improved cancer cell targeting by synergistically enhancing ROS generation and reducing normal cell susceptibility. These findings underscore Al-ZnO NPs as tunable, selective therapeutic agents, leveraging dopant-driven redox modulation to optimize oncological efficacy while sparing healthy tissue.

Ferroptosis is a form of iron-dependent cell death of interest for the development of novel anti-cancer therapies. Ferroptosis research uses a process of elimination based on assumed ferroptosis-specific inducers and inhibitors; these molecules however have off-target effects and cannot provide a comprehensive picture of overlapping pathways. We investigated whether pyroptosis-a form of inflammatory cell death-is initiated in cancer cells following treatment with the ferroptosis inducer RSL3. We treated 6 cancer cell lines with RSL3 alone or in combination with inhibitors of ferroptosis (Ferrostatin-1), caspases (zVADfmk), necroptosis (Necrostatin-1), BID (BI-6C9), or STING (H-151). Biomarkers of pyroptosis and ferroptosis were assessed using our novel quantitative multiplex immunoassay. Increased secretion of pyroptosis-associated cytokines (IL-1α, IL-1β, IL-18), and gasdermin D and E (GSDMD/E) cleavage with parallel loss of respective full-length proteins-both hallmarks of pyroptosis-were recorded in 5/6 cell lines following RSL3 treatment. RSL3 cytotoxicity was blocked by Ferostatin-1; BID and STING inhibitors also prevented GSDMD/E cleavage. We conclude that the ferroptosis-inducer RSL3 triggers pyroptosis in cancer cells; further work is required to elucidate the role of mitochondria in this process. Measurement of pathway-specific protein biomarkers is therefore necessary to identify the exact mechanism of action of novel cytotoxic agents.

Hilar cholangiocarcinoma (hCCA), a rare cancer of the biliary system, has a poor prognosis. This study aimed to investigate the risk factors affecting the survival of patients with hCCA after curative-intent resection and establish a survival predictive model. Clinical data from 340 hCCA patients who underwent curative-intent resection at the First Affiliated Hospital of Xi'an Jiaotong University between 2010 and 2021 were collected. The patients were randomly assigned to a training set and a testing set in a 7:3 ratio. Risk factors selection was performed by five machine learning (ML) algorithms, including Least Absolute Shrinkage and Selection Operator (LASSO) Regression, Forward Stepwise Cox regression, Boruta feature selection, Random Forest and eXtreme Gradient Boosting (XGBoost). A nomogram was constructed based on identified risk factors. The independent risk factors for the postoperative survival in hCCA patients included positive margin, lymph node metastasis, low total lymph node count (TLNC) and poor tumor differentiation. In the training and testing sets, the consistency index (C-index) of ML-based nomogram was 0.731 (95% CI: 0.684-0.753) and 0.714 (95% CI: 0.661-0.775), while the 3-year AUC of the nomogram was 0.784 (95% CI: 0.724-0.844) and 0.770 (95% CI: 0.763-0.867), respectively. The calibration curves for the nomogram showed good concordance. Based on the decision curve analysis, the nomogram had a good clinical application value, outperforming both the TNM staging system and the Bismuth-Corlette classification. Furthermore, patients were stratified into three groups with varying risks of overall survival (OS): the low-risk, middle-risk and high-risk group according to the nomogram, with statistically significant differences observed among these groups (p < 0.001). The ML-based nomogram provided a personalized prognostic prediction model for hCCA patients after surgical resection.

Low-grade glioma(LGG) is a prevalent primary brain tumor, and type I interferons(IFN-Is) can exert a multifaceted influence on the regulation of the tumor microenvironment during its initiation and progression. To investigate the role of IFN-Is in the progression of LGG, we screened public databases for features based on IFN-alpha(IFN-α) response genes (IRGs), a risk model was constructe and its relationship to tumor immunity and prognosis was evaluated. In addition, the expression and regulation of IRGs in LGG were further studied in vitro biological function experiments and immunohistochemistry of clinical samples. 20 differential genes associated with LGG-IRGs were found, then 6-IRGs-based signature was found by using of univariate COX, LASSO and SVM-RFE. ROC curves also supported the value of signature. CIBERSORT results demonstrated the crucial role played by these key signature genes in immune response. Additionally, aggregation analysis of relevant immune-related genes revealed that cluster 1 exhibited the lowest expression of RIPK2 and SELL. Moreover, GSVA results suggested that diagnostic immune-related genes may regulate LGG by influencing immune cells. We further validated these findings using external datasets and provided additional evidence supporting the predictive value of signature genes associated with IRGs in LGG through immunohistochemical testing on clinical samples and in vitro experiments. These 6 diagnostic IRGs genes can help differentiate and predict the prognosis and immune status of patients with LGG, thereby providing new strategies for precise and personalized immunotherapy.

The brain tumours originate in the brain or its surrounding structures, such as the pituitary and pineal glands, and can be benign or malignant. While benign tumours may grow into neighbouring tissues, metastatic tumours occur when cancer from other organs spreads to the brain. This is because identification and staging of such tumours are critical because basically all aspects involving a patient's disease entail accurate diagnosis as well as the staging of the tumour. Image segmentation is incredibly valuable to medical imaging since it can make possible to simulate surgical operations, diseases diagnosis, anatomical and pathologic analysis. This study performs the prediction and classification of brain tumours present in MRI, a combined classification and localization framework model is proposed connecting Fully Convolutional Neural Network (FCNN) and You Only Look Once version 5 (YOLOv5). The FCNN model is designed to classify images into four categories: benign - glial, adenomas and pituitary related, and meningeal. It utilizes a derivative of Root Mean Square Propagation (RMSProp)optimization to boost the classification rate, based upon which the performance was evaluated with the standard measures that are precision, recall, F1 coefficient, specificity and accuracy. Subsequently, the YOLOv5 architectural design for more accurate detection of tumours is incorporated, with the subsequent use of FCNN for creation of the segmentation's masks of the tumours. Thus, the analysis proves that the suggested approach has more accuracy than the existing system with 98.80% average accuracy in the identification and categorization of brain tumour. This integration of detection and segmentation models presents one of the most effective techniques for enhancing the diagnostic performance of the system to add value within the medical imaging field. On the basis of these findings, it becomes possible to conclude that the advancements in the deep learning structures could apparently improve the tumour diagnosis while contributing to the finetuning of the clinical management.

Breast cancer, a heterogeneous disease, is notable for its high morbidity, recurrence, and mortality. T3N0M0 is controversial in terms of clinical prognosis and therapeutic decision-making. We collected baseline and treatment information on patients with T3N0M0 between 2010 and 2015 from the Surveillance, Epidemiology, and End Results programme (SEER) database. Univariate analyses of clinicopathological characteristics were performed using the log-rank test, while variables that were statistically significant (p < 0.05) were included in multivariate analyses to identify prognostic risk factors included in the construction of nomograms. Receiver operating characteristic (ROC) curves, calibration curves and decision curve analysis (DCA) curves were applied to evaluate the nomogram. We obtained a total of 4,641 patient clinical characteristics from the SEER database. In T3N0M0 group, the overall survival (OS) for 3, 5 and 10 years was 87.1%, 79.5% and 71.6%, and breast cancer-specific survival (BCSS) was 92.0%, 88.1% and 85.3%, respectively. We ultimately identified eight variables (age, grade, surgery, radiation, chemotherapy, ER-status, PR-status, Her2-status) were associated with OS while seven variables (age, grade, surgery, radiation, chemotherapy, PR-status, Her2-status) were associated with BCSS. The nomogram was constructed based on independent prognostic risk factors, the area under the curve (AUC) values were well performed, the ROC curves, calibration curves and DCA curves all indicated that the nomogram had better clinical practicability. The current study provides a new clinical assessment tool for T3N0M0 breast cancer patients, as well as a reference for clinicians in selecting management decisions.

Autophagy plays critical and complicated roles in tumors. As the central hub of nutrient signaling and cell growth, mTOR constitutes mTORC1 to be the main gateway for modulating autophagy. Yet, the regulatory mechanisms of mTORC1-regulated autophagy in tumors are not fully deciphered. Here, we report a novel long noncoding RNA, LINC00622, which modulates mTORC1-regulated autophagy in cutaneous melanoma. Functionally, LINC00622 acts as a pro-oncogenic factor to promote proliferation, colony formation, migration and invasion in melanoma while suppressing cell death. Mechanistically, LINC00622 associates with and recruits BTF3 to transcriptionally enhance RRAGD expression for activating mTORC1 and thus inhibiting autophagic cell death, which contributes to the development of cutaneous melanoma. Our findings not only demonstrated the oncogenic role of LINC00622 via RRAGD/mTORC1 axis to repress autophagic cell death in cutaneous melanoma, but also offer novel treatment targets for melanoma therapy.

Up to 40% of diffuse large B-cell lymphoma (DLBCL) patients do not experience a durable response to frontline immunochemotherapy, and prospective identification of high-risk cases that may benefit from personalized therapeutic management remains an unmet need. Molecular phenotyping techniques have established a landscape of genomic variants in diagnostic DLBCL; however, these have not yet been applied in large-scale studies of relapsed/refractory DLBCL, resulting in incomplete characterization of mechanisms driving tumor progression and treatment resistance. Here, we performed an integrated multiomic analysis on 228 relapsed/refractory DLBCL samples, including 24 with serial biopsies. Refined cell-of-origin subtyping identified patients harboring GCB and DZsig+ relapsed/refractory tumors in cases with primary refractory disease with remarkably poor outcomes, and comparative analysis of genomic features between relapsed and diagnostic samples identified subtype-specific mechanisms of therapeutic resistance driven by frequent alteration to MYC, BCL2, BCL6, and TP53 among additional strong lymphoma driver genes. Tumor evolution dynamics suggest innate mechanisms of chemoresistance are present in many DLBCL tumors at diagnosis, and that relapsed/refractory tumors are primarily comprised of a homogenous clonal expansion with reduced tumor microenvironment activity. Adaptation of personalized therapeutic strategies targeting DLBCL subtype-specific resistance mechanisms should be considered to benefit these high-risk populations.

Tyrosinase (TYR) catalyzes the two initial steps of melanin synthesis from tyrosine in various organisms. However, overproduction, accumulation, and abnormal reduction of melanin can lead to severe diseases, particularly skin diseases, which makes tyrosinase a significant endogenous target in developing therapeutics to treat melanin-associated disorders. Herein, we devise a TYR-based in situ catalytic platform that can generate drugs intracellularly through an endogenous copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. By taking advantage of the potent catalytic activity of TYR that is mechanistically validated by ab initio molecular dynamics (AIMD) theoretical calculation and experimental catalysis performance, we develop a TYR-catalyzed in-situ formed proteolysis-targeting chimeras (PROTACs) to degrade intracellular TYR protein to decrease melanin synthesis for treating hyperpigmentation and a TYR-catalyzed in-situ activated prodrug strategy to overcome drug resistance for melanoma therapy. In male mouse models, we show that this TYR-catalyzed therapeutics could efficiently alleviate skin hyperpigmentation within 48 h as well as resensitize the drug-resistant melanoma cells to chemotherapeutics to control tumor growth. Together, we offer an integrative platform to leverage the catalytic activity of endogenous TYR to generate therapeutics through in situ bioorthogonal chemistry for treating melanin-associated skin diseases.

The m6A methyltransferase METTL3 is a key regulator of RNA m6A modification, which plays a critical role in cancer development. Despite the significance of METTL3 in hepatocellular carcinoma (HCC), its post-translational modifications and their functional implications in HCC remain poorly understood. The present study reveals that METTL3 undergoes O-GlcNAcylation, which enhances its stability and promotes HCC progression. Specific O-GlcNAcylation sites (T186/S192/S193) in METTL3 are identified. O-GlcNAc modification reduces METTL3 ubiquitination, thereby increasing protein stability, and enhances its interaction with WTAP, thereby sustaining m6A levels in hepatoma cells. Notably, METTL3 O-GlcNAcylation upregulates the expression of minichromosome maintenance protein 10 (MCM10) by stabilizing its mRNA via an m6A-IGF2BP3-dependent manner. Targeting METTL3 O-GlcNAcylation with designed peptides effectively inhibits HCC growth both in vitro and in vivo. Collectively, our findings provide insights into the regulatory role of O-GlcNAcylation in modulating the m6A epitranscriptome and suggest the potential therapeutic relevance of targeting METTL3 O-GlcNAcylation in HCC.

OTU deubiquitinase 6B (OTUD6B) study in tumors is gradually increasing; however, studies on the role of OTUD6B in colorectal cancer (CRC) are rare. OTUD6B was overexpressed in some human CRC and liver metastasis samples. Although OTUD6B facilitated migration and invasion in CRC cells, it exhibited opposite effects on liver metastasis in immunodeficient and immunocompetent mice. We demonstrated that Otud6b enhanced metastasis in nude mice, but it recruited more CD8+ T cell infiltration in colorectal liver metastasis (CRLM) mouse model of C57BL/6J to inhibit CRLM through upregulating Cxcl11. Furthermore, we demonstrated that OTUD6B deubiquitinated and stabilized DDX5. Ectopically expressed DDX5 facilitated transcription factor STAT3 activation by resolving the RNA G-quadruplex structure of STAT3, resulting in a higher level of CXCL11 transcription and an increase in tumor-infiltrating CD8+ T cells. All-trans retinoic acid inhibited CRLM by upregulating OTUD6B.

Cancer is characterized by chromosomal instability (CIN), which leads to tumor heterogeneity and other malignant features. CIN is caused by abnormal centromere and kinetochore function, which results in aneuploidy, rearrangements, and micronucleus production. Centromere and kinetochore gene misexpression plays a vital role in tumor progression. Here we show that Centromere Protein T (CENPT) is highly expressed in renal carcinoma (RCC) and promotes the tumor proliferation and metastasis of RCC. CENPT is found to be critical for regulating the glutathione (GSH) metabolism pathway because it interacts with γ-glutamyl-cysteine ligase catalytic subunit (GCLC), consequently reducing reactive oxygen species levels and inhibiting ferroptosis. Mechanistically, CENPT increases the catalytic activity of GCLC by directly binding to GCLC ∆213-424aa competitively with glutamate-cysteine ligase modifier subunit (GCLM), consequently induces the GSH synthesis. In turn, GSH increases CENPT expression via transcriptional regulation mediated by the transcription factor ATF2, forming a CENPT-GCLC-GSH feedback loop that enhances the pro-carcinogenic effect of this axis in RCC. Our study identifies CENPT a potential target for RCC via forming a CENPT-GCLC-GSH feedback loop to inhibit ferroptosis. This may support a promising treatment strategy for RCC.

In vascular diseases, the perturbation of key metabolic pathways is extensively studied and recognized as both a causal factor and a consequence in disease onset and progression. Similarly, the dysregulation of the transcriptional landscape within vascular unit components (endothelial and vascular smooth muscle cells) is a pivotal aspect of disease pathogenesis. Metabolite-induced post-translational modifications of transcription factors significantly modulate their functionality, leading to drastic changes in transcriptional outcomes. This review offers a new perspective by elucidating the link between metabolic alterations and metabolite-induced post-translational modifications of transcription factors integral to the pathogenesis of four paradigmatic vascular pathologies: atherosclerosis, diabetes, pulmonary hypertension and tumor angiogenesis.

p14ARF is a lysine-less tumor suppressor that enhances SUMOylation of its interactors. Although p14ARF is known to interact with the E2 SUMO conjugating enzyme UBC9, the link between ARF and SUMOylation is poorly understood and the potential impact of SUMOylation on p14ARF is unknown. Here we show that SUMO2 conjugates to the N-terminus of p14ARF and stabilizes it. Either depleting UBC9 or pharmacologically inhibiting SUMOylation, induces p14ARF degradation. In contrast, blocking ubiquitination or NEDDylation, with TAK-243 or MLN4924/Pevonedistat respectively, increases p14ARF SUMOylation and restores p14ARF levels when SUMOylation is blocked. Treatment with MLN4924 also causes p14ARF-dependent mRNA upregulation of the SUMOylation components SUMO1, SUMO2, and UBC9, globally augmenting SUMOylation. Finally, p14ARF contributes to MLN4924-driven cytotoxicity of prostate cancer cells. Our results provide evidence that, despite lacking lysine, p14ARF is SUMOylated and this modification is critical to counter ubiquitin driven degradation and establishes a new link between inhibition of NEDDylation and SUMOylation.

Alpha-fetoprotein (AFP) is a classic biomarker for hepatocellular carcinoma (HCC). AFP-positive HCC (AFP+ HCC) has been intensively investigated; however, the genomic, transcriptomic and microenvironmental characteristics of AFP-negative HCC (AFP- HCC) remain to be deciphered. Here we show that tumors display mild differences in genetic alterations between AFP- HCC and AFP+ HCC patients, while AFP- HCC exhibits hyperactive arachidonic acid metabolism. Furthermore, the transcription activity of androgen receptor (AR) is significantly increased in AFP- HCC and plays a positive regulatory role in arachidonic acid metabolism and its metabolite 11,12-epoxyeicosatrienoic acid (11,12-EET). The tumor-derived 11,12-EET exhibits high affinity for EGFR that promotes the migration and tube formation of endothelial cells in vitro. Combination of lenvatinib and bicalutamide (an AR antagonist) enhances the therapeutic efficacy for AFP- HCC. Overall, we uncover the AR-mediated hyperactive arachidonic acid metabolism in AFP- HCC, and reveal AR-11,12-EET-EGFR axis-induced angiogenesis, providing a promising strategy of combined AR antagonist with lenvatinib for AFP- HCC treatment.

Aberrant glucose metabolism, a characteristic of malignant tumors, contributes to the development and progression of bladder cancer (BCa). However, the underlying mechanism by which aberrant glucose metabolism promotes BCa progression is still incompletely understood. Here, we demonstrate that low levels of STUB1 are associated with worse progression and poor prognosis of BCa patients. STUB1 overexpression attenuates BCa cell proliferation, migration and amino acid metabolism, especial aspartate metabolism. Mechanistically, we identify that STUB1 induces K6- and K48-linked polyubiquitination of GOT2 at K73 lysine residue to decrease its stability, which attenuates mitochondrial aspartate (Asp) synthesis and regulates mitochondrial dysfunction. GOT2 was significantly up-regulated in BCa tissues and negatively associated with STUB1 expression. Furthermore, we reveal that high glucose stress promotes Asp synthesis and tumor growth through STUB1-GOT2 axis. Collectively, our findings identify that STUB1-GOT2 axis is an important regulator for maintaining Asp synthesis and mitochondrial function in BCa cell growth, which highlights that targeting STUB1-GOT2 axis could be a valuable strategy to ameliorate BCa progression by inhibiting amino acid metabolic function.