ObjectiveTo evaluate the clinical efficacy and safety of toripalimab combined with albumin-bound paclitaxel and cisplatin/carboplatin as first-line treatment for recurrent or metastatic head and neck squamous cell carcinoma (R/MHNSCC).MethodsThirty-five patients with advanced R/MHNSCC admitted to Hunan Cancer Hospital (January 2021-December 2023) received first-line treatment with toripalimab plus albumin-bound paclitaxel and cisplatin/carboplatin. Efficacy was assessed using RECIST 1.1, and adverse events were evaluated according to NCI-CTCAE 5.0.ResultsA total of 35 patients were assessed for efficacy, with partial response (PR) in 21 (60.0%), stable disease (SD) in 6 (17.1%), and progressive disease (PD) in 8 (22.9%). The overall response rate (ORR) was 60.0%, and the disease control rate (DCR) was 77.1%. The follow-up period concluded on August 10, 2024, with a median follow-up duration of 22.0 months (range: 15.0-27.0 months). Among the cohort, 8 patients experienced PD, and 7 patients succumbed to the disease; 13 patients continued treatment without disease progression. The median progression-free survival (PFS) for the entire cohort was 7.0 months (95% CI: 4.4-9.6 months), while the median overall survival (OS) had not yet been reached. Further results of Cox proportional hazards regression analysis showed that programmed death-ligand 1 (PD-L1) expression status was an independent risk factor for progression-free survival (PFS), whereas bone metastasis status and a history of surgery for head and neck squamous cell carcinoma were key risk factors for overall survival. The primary adverse reactions observed included bone marrow suppression, hypothyroidism, rash, neurotoxicity, pneumonia, and abnormal liver function. The majority of patients experienced grade I to II adverse reactions; however, one patient (4.3%) exhibited grade III adverse reactions, specifically an immune-related rash, and two patients experienced grade IV adverse reactions, one with leukopenia and the other with neutropenia. Notably, there were no deaths attributable to toxicity.ConclusionToripalimab combined with albumin-bound paclitaxel and cisplatin/carboplatin demonstrates promising efficacy with manageable adverse reactions as first-line treatment for R/MHNSCC. However, further research involving expanded sample sizes and randomized controlled trials is warranted to substantiate these findings.

4004 Background: Alveltamig (ZG006) is an innovative trispecific T cell engager (Tri-TCE) targeting two distinct DLL3 epitopes on tumor cells and CD3 on T cells (DLL3/DLL3/CD3), designed to bridge tumor cells and T cells, thereby mediating T cell specific killing of tumor cells. Here we conducted a dose optimization study in refractory NEC. Methods: This is a randomized, multicenter, open-label phase 2 study evaluating ZG006 in NEC patients (pts) who failed at least one prior line of therapy. Pts were randomized 1:1 to receive ZG006 at either 10 mg or 30 mg every two weeks, following a priming dose of 1 mg. The primary endpoint is objective response rate (ORR) assessed by IRC per RECIST1.1. Results are reported for 64 pts who received at least one dose of ZG006. Biomarker based efficacy analyses were performed based on a threshold of ≥50% of tumor cells stained at any intensity for DLL3 with an investigational antibody against DLL3 (SP347, Roche Diagnostics). Results: As of Sep. 10, 2025, a total of 64 pts (32 at each dose group) were randomized and received at least one dose of ZG006. The median age was 56 years (range: 25-72), 35 (54.7%) were male. All pts had received ≥1 prior line of systemic therapy, with 65.6% had prior anti-PD-(L)1 treatment. The confirmed ORR in 10 mg and 30 mg groups were 21.9% (7/32) and 37.5% (12/32), respectively; Disease control rate (DCR) were 40.6% (13/32) and 62.5% (20/32), respectively. Among the 19 responders, primary tumor sites included cervix (8), gallbladder (4), stomach (3), rectum (3) and colon (1). Pts with ≥50% DLL3 staining in tumor cells had greater ORR, DCR, and progression-free survival (PFS). The confirmed ORR in 10 mg and 30 mg groups were 33.3% (6/18) and 56.3% (9/16), respectively; DCR were 50.0% (9/18) and 75.0% (12/16), respectively; With a median follow up of 6.4 months, median PFS were 3.02 and 8.41 months, respectively. Median DoR was not yet mature in both groups at data cut-off. Treatment-related adverse events (TRAEs) occurred in all pts. The most frequent events were pyrexia, cytokine release syndrome (CRS) and anaemia. Most CRS were Grade 1-2, with only four grade 3 CRS, resolved with supportive care. TRAEs led to treatment interruption in 15 pts (23.4%) and permanent discontinuation in 2 pts (3.1%, one in each dose group). No grade 5 TRAE. No significant difference was observed in the safety profile between these two dose groups. Conclusions: ZG006 demonstrated a robust and durable response and a manageable safety profile in previously treated NEC pts, with superior efficacy observed at the 30 mg dose. The confirmed ORR of 56.3% and median PFS of 8.41 months in pts with ≥50% DLL3 positive tumor cells are particularly encouraging, and support further development of ZG006 for this patient population at the 30 mg dose. Clinical trial information: NCT06440057 .

Intracerebral hemorrhage (ICH) is a neurological disorder associated with high mortality and disability rates and can lead to severe neurological injury. This study investigated the role and underlying mechanism of FGD5-AS1 in ICH. In vitro and in vivo models of ICH were established. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to measure FGD5-AS1 and miR-93-5p expression. Neurological deficits were assessed using the modified Neurological Severity Score (mNSS), and cerebral edema was evaluated using the dry-wet weight method. Enzyme-linked immunosorbent assay (ELISA) was used to measure inflammatory cytokines and oxidative stress markers. Cell viability was evaluated using a cell viability and proliferation assay kit. The interaction between FGD5-AS1 and miR-93-5p was examined using dual-luciferase reporter assays and Pearson correlation analysis. Experimental ICH was accompanied by elevated FGD5-AS1 expression in both animal and cellular models. Suppression of FGD5-AS1 alleviated brain swelling, improved neurological performance, reduced inflammatory and oxidative injury, and enhanced endothelial cell survival. Inhibition of miR-93-5p diminished these protective effects, supporting a regulatory interaction between the two molecules. FGD5-AS1 negatively regulated miR-93-5p expression, and inhibition of miR-93-5p attenuated the effects observed following FGD5-AS1 knockdown. These results indicate that FGD5-AS1 may contribute to ICH-associated brain injury through regulation of miR-93-5p and that the FGD5-AS1/miR-93-5p axis may represent a potential target for further investigation in experimental ICH.

Major depressive disorder (MDD) is associated with astrocytic pyroptosis, but the mechanisms involved remain unclear. This study investigates how O-GlcNAc modification of CCAAT enhancer binding protein beta (CEBPB) influences its ability to regulate pyroptosis in astrocytes. In our study, primary astrocytes were exposed to LPS plus ATP to activate NLRP3 inflammasomes. Cell viability was assessed by CCK8 assay. IL-18 and IL-1β release was measured using ELISA. sWGA pull-down and Co-IP assays were performed to assess O-GlcNAc modification level of CEBPB. ChIP assay was used for promoter binding analysis, and ubiquitination detection was performed using Co-IP. Our results showed that LPS+ATP treatment significantly increased IKKβ expression in astrocytes, and IKKβ knockdown markedly reduced NLRP3 inflammasome activity and pyroptosis. Mechanistically, CEBPB transcriptionally activated IKKβ expression in astrocytes. Functionally, CEBPB knockdown ameliorated LPS+ATP-induced NLRP3 inflammasome activation and pyroptosis in astrocytes, which was abolished by IKKβ overexpression. In addition, O-GlcNAc modification at Thr235 enhanced CEBPB protein stability in astrocytes, driving NLRP3 inflammasome activation and pyroptosis. Collectively, these findings demonstrated that O-GlcNAc-modified CEBPB promoted pyroptosis in astrocytes during MDD progression by activating NLRP3 inflammasome through transcriptionally activating IKKβ.

Lung adenocarcinoma (LUAD) remains a leading cause of cancer mortality. This study identifies FXYD6 as a novel tumor suppressor in LUAD, where it is significantly downregulated. High FXYD6 expression correlates with improved patient survival, and its overexpression inhibits LUAD cell proliferation and migration. Mechanistically, FXYD6 is directly targeted and suppressed by the oncomiR miR-4735-3p, which is upregulated in LUAD. Inhibiting miR-4735-3p elevates FXYD6 levels and suppresses malignant phenotypes. Furthermore, we delineate a complete competing endogenous RNA (ceRNA) axis: the long non-coding RNA OIP5-AS1, which is downregulated in LUAD, functions as a molecular sponge for miR-4735-3p. Overexpression of OIP5-AS1 sequesters miR-4735-3p, thereby derepressing FXYD6 expression and inhibiting cancer progression. Rescue assays confirm that miR-4735-3p overexpression reverses the tumor-suppressive effects of OIP5-AS1. In vivo xenograft models validate that the OIP5-AS1/miR-4735-3p/FXYD6 axis critically regulates tumor growth. Our findings reveal a crucial tumor-suppressive pathway in LUAD, highlighting the long noncoding RNA (lncRNA) OIP5-AS1 as a key upstream regulator that modulates FXYD6 by competitively binding miR-4735-3p. This axis presents a promising therapeutic target for LUAD intervention.

Major depressive disorder is associated with impaired excitatory synaptic transmission, but the molecular mechanisms linking chronic stress to altered AMPA receptor trafficking remain incompletely understood. Here we show that chronic mild stress increases OGT-mediated O-GlcNAcylation of PIN at serine 88, which stabilizes PIN and enhances its interaction with nitric oxide synthase. This suppresses nitric oxide synthase activity, reduces stargazin S-nitrosylation, weakens stargazin-GluA1 binding, and impairs GluA1-containing AMPA receptor trafficking. Genetic or pharmacological inhibition of OGT restores this signaling pathway and alleviates stress-induced depression-like behaviors in mice. These findings identify the OGT-PIN-NOS-stargazin axis as a regulator of stress-induced synaptic dysfunction and suggest that targeting OGT may help restore AMPA receptor trafficking in depression-related conditions.

Cervical cancer stands as a prevalent gynecological malignancy. The purpose of this study is to discover new therapeutic targets for cervical cancer. To identify key genes associated with cancer, we analyzed transcriptome expression data of various cancers from TCGA. CCK-8 assays, colony formation assays, wound-healing assays, transwell assays and apoptosis assays were used to investigate their functions. qRT-PCR and Western Blot were utilized to verify their expression. RESULTS: A total of 358 common key genes were identified in six common cancers. Immunohistochemical experiments revealed that FEN1 was significantly overexpressed in cervical cancer. Compared to the control group, knocking down FEN1 markedly inhibited the proliferation, colony formation, migration and invasion capabilities of SiHa and HeLa, while simultaneously promoting apoptosis. In both SiHa and HeLa cells, knockdown of FEN1 significantly reduced the expression of PCNA. PCNA knockdown exerted nearly identical inhibitory effects on the malignant phenotypes of these two cell lines as FEN1 depletion. Subsequent rescue experiments verified that restoring PCNA expression could partially reverse the suppressive effects of FEN1 knockdown on the malignant behaviors of cervical cancer cells. Moreover, FEN1 silencing in both SiHa and HeLa cells also led to a significant decrease in the expression levels of BCL-2, RB1 and PIK3CA, while increasing the expression of Caspase-9. CONCLUSION: FEN1 exhibits a significant overexpression in cervical cancer and has the capacity to modify the biological behavior of cancer cells, potentially via regulating the expression of PCNA. Inhibiting FEN1 expression may slow down the progression of cervical cancer.

Neuroinflammation is caused by the overactivation of microglia, contributing to secondary brain injury in ischemic stroke. Oxytocin (OXT), a neuropeptide synthesized by neurons in the paraventricular nucleus (PVN) of the hypothalamus, has demonstrated potential in mitigating inflammatory responses across various pathological conditions. However, research on the role of PVNOXT neurons in ischemic stroke is limited, and the modulatory effect of these neurons on neuroinflammation remains unclear. Transient middle cerebral artery occlusion (tMCAO) was performed in mice. OXT levels were measured in the peri-infarct cortex, serum, and cerebrospinal fluid (CSF), and OXTR expression was assessed in the peri-infarct cortex after tMCAO. Chemogenetic approaches were used to selectively activate PVNOXT neurons in OXT-Cre mice, after which neurological function, infarct volume, and blood–brain barrier integrity were evaluated. To investigate the mechanisms underlying the regulation of ischemic injury by PVNOXT neurons, RNA sequencing of the ipsilateral ischemic hemisphere was performed. Additional analyses, including flow cytometry, molecular assays, and Transwell migration experiments, were conducted to validate the downstream signaling pathways and cellular responses. OXT levels significantly reduced in the peri-infarct cortex, serum, and CSF, whereas OXTR expression increased in the peri-infarct cortex after tMCAO. Chemogenetic activation of PVNOXT neurons increased OXT levels in both the brain and circulation, reduced infarct volume, and improved neurological outcomes. In addition, transcriptomic analysis identified CXCL3 as one of the most significantly downregulated chemokines after the activation of PVNOXT neurons, and CXCL3 downregulation was associated with reduced neutrophil chemotaxis. Further in vivo and in vitro investigations demonstrated that PVNOXT neurons inhibit microglial CXCL3 expression via the OXTR–ERK signaling pathway, thereby restricting the infiltration of neutrophils. In contrast, the administration of recombinant CXCL3 promoted the recruitment of neutrophils and exacerbated ischemic injury. PVNOXT neurons alleviate post-ischemic brain injury by inhibiting the secretion of CXCL3 from microglia, consequently reducing neutrophil chemotaxis. These results underscore the therapeutic potential of targeting PVNOXT neurons and their downstream signaling pathways to mitigate immune-mediated damage in ischemic stroke.

Immune checkpoint inhibitors have been proven effective for recurrent or metastatic cases of microsatellite instability-high (MSI) endometrial cancer (EC). However, drug resistance exists in a noticeable proportion of patients. Elucidating the underlying mechanisms would help develop new therapeutic strategies and benefit in improving patients' prognosis. Circular RNAs (circRNAs) are excellent biomarkers due to their stability and tissue specificity. Evidence has showed that circRNAs could mediate immune evasion in several types of malignancies. However, whether they regulate the immune response in MSI EC has not been explored. Here, based on the results of our former circRNA array, which identified the differentially-expressed circRNAs in MSI EC, we found that a circRNA, circCLMP, was negatively correlated with CD8+ T cell infiltration in MSI EC, and up-regulated in ICI-resistant MSI EC. In vivo assays showed that circCLMP could alter the anti-tumor immunity and promote tumor growth. Mechanistically, circCLMP shielded IRF3 from binding to TBK1, interfered with the phosphorylation and nuclear translocation of IRF3, thereby inhibiting the activation of interferon response, suppressing CD8+ T cell infiltration in the tumor environment, and eventually mediating immune evasion and promoting the progression of MSI tumors. Targeted knockdown of circCLMP combined with anti-PD-1 inhibitor treatment effectively enhanced the anti-tumor effects in the preclinical MSI EC PDX model. For the first time, our study reported an immunoregulating circRNA in MSI EC, which may provide insights into developing new biomarkers and therapeutic targets for overcoming immunotherapy resistance in MSI EC.

BACKGROUND:Endometrial carcinoma (EC) is a prevalent kind of cancerous tumor with significant morbidity and mortality. Mounting evidence reveals that lactylation modification plays a crucial role in tumorigenesis, but its connection to EC remains poorly understood. This study aimed to identify a lactylation-related gene signature to predict the prognosis of EC. METHODS:Differentially expressed lactylation-related genes between EC and normal samples were analyzed using the TCGA database. Univariate and LASSO Cox regression analyses were employed to construct the lactylation-related signature, which was then validated using both the test set and entire set. A nomogram was further developed and evaluated. Additionally, enrichment analysis, immune cell infiltration, tumor mutation burden and drug response were assessed between the two risk groups. RESULTS:Sixteen lactylation-related genes (LRGs) were selected to construct the prognostic signature. Kaplan-Meier survival curves showed that patients in the high-risk group had remarkably worse prognosis. A nomogram based on the signature and other clinical characteristics was constructed and demonstrated strong predictive power. Additionally, biological pathways, immune status, tumor mutation burden and drug response differed between the high- and low-risk groups. CONCLUSION:In conclusion, our study demonstrated that the LRG signature is a promising biomarker for EC, effectively distinguishing high-risk patients, predicting prognosis, and offering new strategic directions for antitumor immunotherapy.

The lymph node is the most common site of distant metastasis of cervical cancer (CCa), which elicits dismal prognosis and limited efficiency for treatment. Identification of the factors contributing to CCa lymphatic metastasis is needed to develop effective prevention and treatment strategies. Here, we found upregulation of prolyl 4-hydroxylase subunit alpha 3 (P4HA3), an α-subunit of prolyl hydroxylase, in lymphatic metastatic lesions of cervical cancer, which is strongly associated with poor prognosis. In vitro and in vivo experiments showed that P4HA3 promoted CCa lymphatic metastasis by conferring ATP-citrate lyase (ACLY)-mediated ferroptosis resistance. Mechanistically, P4HA3 stabilizes ACLY protein by competitively inhibiting its interaction with the E3 ubiquitin ligase UBR4, which prevents UBR4-mediated proteasomal degradation of ACLY. ACLY-derived acetyl-CoA enhances H3K27 acetylation (H3K27ac) modification level in the promoter of SLC7A11 gene, ultimately enhancing SLC7A11 transcription and ferroptosis resistance. Collectively, our study provides a mechanistic understanding of the interplay between ferroptosis resistance and lymph node metastasis, providing a possibility to combat lymph node metastasis in cervical cancer.

Lymph node metastasis (LNM) confers significant treatment failure and adverse clinical outcomes in cervical cancer (CCa). However, large unknown lies in the mechanisms underlying LNM in CCa. In this study, we discovered that PRDX3 is elevated in CCa with LNM and associates with poor prognosis in CCa patients. Moreover, multivariate logistic analysis revealed that PRDX3 is an independent predictor of LNM in CCa. Functional investigations demonstrated that PRDX3 promotes invasion, lymphangiogenesis, and LNM of CCa. Mechanistically, PRDX3 activates NF-κB signaling pathway and upregulates the downstream pro-metastatic molecules VEGF-C and MMP-9 in CCa cells. Furthermore, PRDX3 could inhibit the anoikis of detached CCa cells by reducing the ROS level and hence promote LNM in CCa. Importantly, genetic inhibition of PRDX3 potently slows LNM of CCa. These findings highlight a novel PRDX3-mediated mechanism of LNM in CCa and recognize PRDX3 as a promising predictive marker and target of clinical intervention for LNM in CCa.

Acute myeloid leukemia is a malignant hematologic disorder characterized by the excessive proliferation and accumulation of immature myeloid cells. This abnormality disrupts normal hematopoiesis, leading to symptoms such as anemia, increased susceptibility to infections and bleeding. ADP-ribosylation factors (ARFs) are critical in various cellular functions, including vesicular trafficking, membrane dynamics, cytoskeleton organization, signal transduction, endocytosis, exocytosis, and maintaining organelle integrity. Among ARF family members, ARF3 has garnered relatively less attention compared to other members like ARF1 and ARF6, leaving its role less understood. In this study, we found that the higher expression of ARF3 is correlated with poorer prognosis in AML patients. Silencing ARF3 in AML cells interrupted cell cycle progression and promote cell death as well as inhibit leukemogenesis in vivo. Mechanically, ARF3 knockdown suppressed AML progression by inhibiting PI3K/Akt signaling pathway. Our results indicate that ARF3 is linked to poor outcomes in AML patients and can serve as a potential therapeutic target for AML treatment.

Lymph node metastasis (LNM), a hallmark of aggressive cervical cancer (CCa), severely compromises prognosis and limits therapeutic efficacy. Despite its clinical significance, the molecular mechanisms driving LNM in CCa remain poorly characterized, necessitating urgent exploration. In this study, leveraging an in vitro lymphangiogenesis screening model, we identified CREB5 as a gene markedly upregulated in metastatic CCa, with its expression strongly correlating with LNM and poor patient survival. Mechanistic investigations revealed that CREB5 transcriptionally activates apelin (APLN) by directly binding to its canonical TGACG motif in the promoter region, thereby driving APLN-mediated lymphangiogenesis. Inhibition of CREB5 or treatment with ML221, a selective APLN receptor (APLNR) antagonist, potently suppressed lymphatic metastasis in preclinical CCa models. These findings establish CREB5 as a critical regulator of APLN-dependent lymphangiogenesis and LNM, highlighting its potential as a dual diagnostic biomarker and therapeutic target for mitigating lymphatic dissemination in CCa patients.

The efficacy of the SGLT2 inhibitor empagliflozin (EMPA) in mitigating hepatic steatosis in patients with type 2 diabetes mellitus and metabolic dysfunction-associated steatotic liver disease (MASLD) has been previously demonstrated. However, the underlying mechanisms remain unclear. In this study, we investigated the role of EMPA in alleviating hepatic steatosis through the modulation of O-GlcNAcylation. High-glucose (HG)-induced alpha mouse liver 12 (AML12) cells, mouse primary hepatocytes (MPHs), and murine MASLD models (high-fat diet-fed and ob/ob mice) were used to examine the effects of EMPA. Protein O-GlcNAcylation, lipid accumulation, and AMP-activated protein kinase α (AMPKα) regulation were evaluated using Western blotting, immunostaining, and siRNA knockdown. Our findings showed that protein O-GlcNAcylation levels were elevated in both in vitro and in vivo models. EMPA treatment reduced O-GlcNAcylation and ameliorated lipid accumulation in HG-induced AML12 cells, MPHs, and MASLD models. Knockdown of O-GlcNAc transferase (OGT) decreased O-GlcNAcylation levels and lipid accumulation in HG-induced AML12 cells. Additionally, OGT knockdown altered both O-GlcNAcylated and phosphorylated AMPKα levels. In these models, EMPA administration decreased O-GlcNAcylated AMPKα while increasing phosphorylated AMPKα. This study further identified serine 344, threonine 447, and serine 501 as critical O-GlcNAcylation sites on AMPKα2. Mutation of these residues to alanine in AMPKα2 attenuated lipid accumulation in AML12 cells, with no additional improvement observed following EMPA treatment. In summary, EMPA effectively improves hepatic steatosis by modulating the O-GlcNAcylation states of AMPKα. Identification of specific O-GlcNAcylation sites on AMPKα2 highlights their importance in the therapeutic mechanism of EMPA in improving hepatic steatosis.

OBJECTIVE:To develop a predictive model for the efficacy of anlotinib combined with osimertinib in treating advanced non-small cell lung cancer (NSCLC) with acquired resistance, based on circulating tumor DNA (ctDNA) clearance and angiogenesis markers. METHODS:We retrospectively analyzed data from 378 patients with acquired resistance advanced NSCLC treated at Shanghai University Mengchao Cancer Hospital between November 2020 and October 2022. Patients were divided into a monotherapy group (n=181, receiving osimertinib alone) and a combination therapy group (n=197, receiving anlotinib plus osimertinib). The combination therapy group was further stratified into two subgroups based on progression-free survival (PFS): better efficacy (PFS ≥16 months) and poorer efficacy (PFS <16 months). Venous blood samples were collected before and after two treatment cycles for ctDNA detection using next-generation sequencing (NGS) and measurement of angiogenesis markers using electrochemiluminescence immunoassays. Multivariate logistic regression was employed to identify independent predictors, and a nomogram predictive model was constructed and validated. RESULTS:The combination therapy group demonstrated significantly longer PFS and overall survival compared to the monotherapy group. After two treatment cycles, the combination therapy group showed a significant reduction in ctDNA levels, variant allele frequency (VAF), and a higher clearance rate of the T790M mutation. Additionally, angiogenesis markers were significantly lower in the combination therapy group post-treatment (all P<0.05). Multivariate logistic regression identified significant predictors of poor therapeutic effect. The nomogram model exhibited excellent predictive performance, with an AUC of 0.912 in the training set and 0.959 in the test set. CONCLUSION:The developed nomogram reliably predicts the efficacy of anlotinib combined with osimertinib in treating acquired resistance NSCLC, offering valuable guidance for personalized treatment decisions.

Negative regulator of ubiquitin-like protein 1 (NUB1), an inhibitor of neural precursor cells expressed developmentally downregulated 8 (NEDD8), is implicated in tumor growth. However, the expression of NUB1 in hepatocellular carcinoma (HCC) and its effects on HCC growth remain unclear. In this study, our findings revealed reduced NUB1 protein expression in HCC tissues and cells, leading to increased proliferating cell nuclear antigen (PCNA) protein stability through upregulating NEDD8 to promote HCC cell growth. Mechanistically, NUB1 reduction upregulated NEDD8 to promote PCNA NEDDylation at lysine 164 (Lys164), in turn, antagonized PCNA K48-linked polyubiquitination, thereby increasing the stability of PCNA in HCC cells. Finally, the results of the in vitro and in vivo experiments revealed that the NEDDylation inhibitor TAS4464 could inhibit PCNA NEDDylation to decrease PCNA protein expression, thereby suppressing HCC cell growth. Collectively, our results identified NUB1 as a negative regulator of HCC proliferation and confirmed that PCNA NEDDylation promotes PCNA protein stability by antagonizing PCNA polyubiquitination. This study provides a new perspective on the specific mechanism of HCC growth. It expands our understanding of the role of NEDDylation in the regulation of substrate proteins and their functions.

Endometrial receptivity is essential for successful pregnancy, and endometriosis is widely recognized as a disruptor of this process. Poor endometrial receptivity is also a key factor contributing to recurrent implantation failure. Although some molecular mechanisms related to endometrial receptivity have been identified, their specific roles in endometriosis and recurrent implantation failure remain unclear. This study aimed to elucidate the shared molecular mechanisms affecting endometrial receptivity in endometriosis and recurrent implantation failure using multi-omics data analysis. We sourced datasets from the NCBI GEO database and employed weighted gene co-expression network analysis to identify gene modules associated with these conditions, followed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Single-cell sequencing analysis and immunofluorescence were used for expression analysis. We identified 3690 and 4892 upregulated genes and 2675 and 5065 downregulated genes in endometriosis and recurrent implantation failure, respectively. Functional enrichment analysis and validation identified 15 hub genes including SRPRB, SLC35B1, and SLC25A6. Receiver operating characteristic curve analysis demonstrated that these genes are associated with high diagnostic accuracy. Single-cell sequencing analysis indicated that these genes are predominantly expressed in basal epithelial cells, with RBM3 being particularly prominent. This study provides new insights into the molecular mechanisms underlying endometrial receptivity and identifies potential targets for the diagnosis and treatment of endometriosis and recurrent implantation failure.