Recent studies have shown that zinc finger protein 750 (ZNF750), a key tumor suppressor in esophageal squamous cell carcinoma (ESCC), is associated with defective epithelial differentiation. At the same time, mucosal immune dysfunction is increasingly recognized as a key factor in the development of ESCC. Exploring the role of ZNF750 in regulating the mucosal immune microenvironment provides a new perspective on its tumor suppression mechanism. By integrating analyses across multiple platforms, including bulk transcriptomics (TCGA, GTEx, GEO), and single-cell RNA sequencing (scRNA-seq), we demonstrate that ZNF750 is significantly downregulated in ESCC, with high expression serving as an independent prognostic marker for longer survival. Mechanistically, ZNF750 suppresses ESCC progression by maintaining epithelial barrier–driven mucosal immune homeostasis. WGCNA analyses indicate that ZNF750-high tumors are enriched in mucosal immune response and antimicrobial defense pathways. Experimental validation reveals that ZNF750 positively regulates DEFB4A (human beta-defensin-2), a key antimicrobial peptide. In addition, ZNF750 fosters an anti-tumor immune environment by recruiting neutrophils and suppressing pro-tumor M2 macrophages, a finding corroborated by clinical validation showing that high ZNF750 expression correlates with elevated neutrophil infiltration. Clinical validation confirms that high ZNF750 expression correlates with elevated neutrophil infiltration. Additionally, ZNF750 expression is linked to a healthy mucosal microbiota and the exclusion of pro-carcinogenic bacteria. Single-cell analysis shows that ZNF750-expressing epithelial cells maintain barrier integrity and coordinate immune surveillance through MHC-I/II signaling. Loss of ZNF750 may drive mucosal immune dysregulation, where the immune network is replaced by a pre-fibrotic immunosuppressive niche dominated by cancer-associated fibroblasts (CAFs). Collectively, our findings identify ZNF750 as a key molecular link between epithelial barrier integrity and mucosal immune surveillance in ESCC, highlighting restoration of ZNF750-mediated barrier immunity as a potential therapeutic strategy for ESCC.

Zinc finger protein 750 (ZNF750) is a novel tumor suppressor in esophageal squamous cell carcinoma (ESCC). However, its role in metabolic remodeling in ESCC remains unclear. Single-cell RNA sequencing data (GSE160269) from 66 ESCC samples indicated that ZNF750 was specifically expressed in epithelial cells and was associated with lipid metabolic pathways in ESCC. Lipidomic assays demonstrated that ZNF750 deficiency induced lipid metabolic reprogramming, especially polyunsaturated fatty acid (PUFA) remodeling, and significantly enhanced ESCC cell membrane fluidity and promoted malignant phenotypes. Further transcriptomic analysis indicated that ZNF750-associated genes were enriched in pathways related to PUFA metabolism. Mechanistically, ZNF750 binds to the promoter of fatty acid desaturase 1 (FADS1) to inhibit its transcription and expression, thereby modulating lipid saturation and ultimately impacting ESCC cell phenotypes. Furthermore, ZNF750 knockdown significantly enhanced the ferroptosis sensitivity of ESCC cells through transcriptional regulation of acyl-CoA synthetase long-chain family member 4 (ACSL4). Overall, our study delineates the lipidomic landscape regulated by ZNF750 and elucidates its dual role in modulating PUFA metabolism and ferroptosis sensitivity. These findings suggest that targeting PUFA metabolism and ferroptosis are potential therapeutic strategies for ESCC patients with ZNF750 mutations or deletions.

BACKGROUND:Esophageal squamous cell carcinoma (ESCC) lacks effective targeted therapies. The tumor suppressor ZNF750 is frequently inactivated, yet the primary mechanism and its therapeutic implications remain poorly defined. METHODS:We performed an integrated multi-omics analysis on a large ESCC cohort (n = 767). Mechanistic insights were validated by ChIP-qPCR, luciferase assays, and functional studies in vitro and in vivo. FINDINGS:ZNF750 was transcriptionally silenced in 93.5% of tumors, and this was primarily driven by highly activated NF-κB signaling rather than genetic alteration. We identified a pathogenic feedback loop: NF-κB1(12-fold enrichment on the ZNF750 promoter) represses ZNF750 transcription, while ZNF750 protein suppresses the expression of CARD14, a potent NF-κB activator (mRNA reduced by >70% upon ZNF750 re-expression). ZNF750 loss thereby licenses constitutive NF-κB signaling. Crucially, ZNF750-deficient tumors exhibited heightened sensitivity to the NF-κB-inhibiting proteasome inhibitor, bortezomib, both in cell lines (IC50) and patient-derived xenograft models. INTERPRETATION:Our findings establish NF-κB-mediated repression as the principal mechanism silencing ZNF750 in ESCC. We define ZNF750 loss as a novel biomarker for patient stratification and provides a strong rationale for the clinical evaluation of bortezomib for this molecularly defined ESCC subtype, offering a mechanistic rationale for a biomarker-driven therapeutic strategy for most ESCC patients.

Background: Immunotherapy efficacy in esophageal squamous cell carcinoma (ESCC) is often limited by an immunosuppressive tumor microenvironment (TME). NLRC5, a key regulator of MHC-I antigen presentation, exhibits context-dependent roles in tumor immunity; however, its function in ESCC remains unclear. This study aimed to systematically investigate the expression pattern, prognostic value, and immunological role of NLRC5 in ESCC. Methods: An integrated analysis of bulk RNA sequencing and single-cell RNA sequencing (scRNA-seq) data was performed using multiple cohorts, including The Cancer Genome Atlas, Gene Expression Omnibus, and an in-house ESCC cohort. Differential expression, survival analysis, immune infiltration estimation, and functional enrichment analyses were conducted to elucidate the role of NLRC5 in the tumor microenvironment. Results: NLRC5 was significantly upregulated in ESCC and its high expression independently predicted poor patient survival. Although NLRC5 expression was associated with increased CD8+ T cell infiltration, it was paradoxically accompanied by features of T-cell exhaustion and elevated expression of multiple immune checkpoints. Moreover, NLRC5-high tumors were enriched in transcriptional programs related to PANoptosis, indicating an additional immunosuppressive mechanism within the TME. Conclusions: NLRC5 is not only a prognostic biomarker but also a key modulator of an immune-active yet functionally suppressed tumor microenvironment in ESCC. These findings highlight NLRC5 as a potential therapeutic target for restoring effective antitumor immunity.

Long non-coding RNAs (lncRNAs) have emerged as crucial regulators of cancer development and progression. Among them, Differentiation Antagonizing Non-Protein Coding RNA (DANCR) has been implicated in various malignancies, including esophageal squamous cell carcinoma (ESCC). This study explores the clinical characteristics, prognostic implications, functional roles, and molecular mechanisms of DANCR in ESCC. Our results demonstrate that DANCR is highly expressed in ESCC, and acts as an oncogene in ESCC both in vitro and in vivo. Through bioinformatics analysis and experimental validation, we revealed that DANCR promotes ESCC progression by sponging miR-3193 and regulating its target gene DDIT3 expression. These findings highlight the critical role of DANCR in the development of ESCC and suggest its potential as a prognostic predictor and drug therapeutic target.

FBXW7 is a tumor suppressor gene that regulates metabolism and is associated with the onset and progression of colorectal cancer (CRC)), however, the precise mechanism whereby FBXW7 participates in the metabolic reprogramming of CRC remains unclear. Here, the research aims to reveal the association between the expression of FBXW7 and clinical variables and to investigate the molecular mechanism by which FBXW7 plays a critical role in the development of CRC. The clinical importance of FBXW7 in CRC was determined by immunohistochemistry. Non-targeted metabolomics was utilized to explore the role of FBXW7 in the metabolic regulation of CRC. Low expression of FBXW7 was associated with poor prognosis in individuals with CRC, both at the mRNA and protein levels. FBXW7 over-expression inhibited CRC cell growth, colony formation, migration, and invasion. Non-targeted metabolomics unveiled that FBXW7 over-expression directly caused the deprivation of arginine which led to downmodulation of mTOR signaling pathway; meanwhile, FBXW7-related metabolites were primarily concentrated in the mTOR signaling pathway. In summary, the research identified a novel mechanism of action of FBXW7 in CRC. The research findings provide a theoretical foundation for the prognostic prediction and therapeutic planning of CRC based on metabolic reprogramming.

Background:Liver fibrosis is characterized by hepatic stellate cell (HSC) activation and collagen overproduction, but its pathogenesis remains largely unknown. This study aimed to uncover the role of neural precursor cell expressed developmentally downregulated 4-like (Nedd4L) signaling in liver fibrosis and its relationship with gut microbiota. Methods:Intraperitoneal injection of carbon tetrachloride (CCl4) was used to induce liver fibrosis in 8-week-old female C57BL/6J mice with Nedd4L knockout or administration of the Nedd4L protein phosphorylation inhibitor EMD638683. HSCs isolated from mice were activated with transforming growth factor-beta 1 (TGFβ1) with or without EMD638683. Results:An approximately 3-fold elevation in Nedd4L mRNA was observed in hepatocytes and liver tissues, and significantly higher hepatic Nedd4L phosphorylation was observed in fibrotic mice than in non-fibrotic mice. Nedd4L mRNA level in HSCs isolated from fibrotic livers and Nedd4L protein level in TGFβ1-stimulated HSCs from wild-type livers remained unchanged. In isolated HSCs, TGFβ1-induced Nedd4L phosphorylation and cell activation were suppressed with EMD638683. In CCl4-treated mice, EMD638683 alleviated liver fibrosis and induced a relative increase in fecal Bacteroides, Parabacteroides, Erysipelatoclostridium, Blautia, and Klebsiella, whereas Nedd4L deficiency predisposed mice to liver injury and liver fibrosis with a remarkable reduction in fecal Lactobacillus, Enterorhabdus, and Bacteroides. Conclusion:Hepatic Nedd4L signaling contributes to CCl4-induced liver fibrosis in female mice, which is associated with alterations in the gut microbiota, and Nedd4L phosphorylation is involved in TGFβ1-mediated HSC activation.

Summary: Tumor extracellular vesicles (EVs) demonstrate considerable promise for medication delivery and tumor targeting owing to their natural long-term blood circulation and tissue targeting capabilities. We extracted EVs from mouse breast cancer cell 4T1 using UV stimulation and differential centrifugation. To create a new nano-drug delivery system, the vesicle delivery system (EPM) loaded with melanin and paclitaxel albumin (PA), the collected EVs were repeatedly compressed on a 200 nm porous polycarbonate membrane with melanin and PA. Our findings suggest that EPM is readily absorbed by breast cancer and dendritic cells. EPM generates significant photoacoustic signals and photothermal effects when exposed to near-infrared light and can enhance the infiltration of CD8+ T cells in mouse tumor tissues. EPM is more cytotoxic than PA in in vivo and in vitro investigations. The efficacy of EPM in clinical transformation when paired with chemotherapy/photothermal/immunotherapy treatment is demonstrated in this study.

As a first-line chemotherapeutic agent, albumin-bound paclitaxel (PA) has a considerable effect on the treatment of various cancers. However, in chemotherapy for hepatocarcinoma, the sensitivity to PA is low owing to the innate resistance of hepatocarcinoma cells; the toxicity and side effects are severe, and the clinical treatment impact is poor. In this study, we present a unique nanodrug delivery system. The ultraviolet (UV)-induced tumor-cell-derived extracellular vesicles (EVs) were isolated and purified by differential centrifugation. Then, PA was loaded by coextrusion to create a vesicle drug delivery system (EVPA). By employing the EV-dependent enhanced retention effect and specific homing effect, EVPA would passively and actively target tumor tissues, activate the immune response to release PA, and achieve the combination therapeutic effect of chemo-immunotherapy on hepatocarcinoma. We demonstrated that the tumor-killing effect of EVPA is superior to that of PA, both in vivo and in vitro and that EVPA can be effectively taken up by hepatocarcinoma and dendritic cells, activate the body’s specific immune response, promote the infiltration of CD4+ and CD8+ T cells in tumor tissues, and exert a precise killing effect on hepatocarcinoma cells via chemo-immunotherapy.

Background: The leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) plays an important role in stem cell differentiation, organ development and cancer. Whether LGR4 affects the progression of hepatocellular carcinoma (HCC) remains unknown. This study aimed to reveal the role of LGR4 in HCC. Methods: Clinical samples of HCC were collected to assess the expression of LGR4 and its correlation with patients' clinical characteristics. The expression level of LGR4 in HCC cells was altered by pharmacological and genetic methods, and the role of LGR4 in HCC progression was analyzed by in vivo and in vitro assays. HCC was induced by diethylnitrosamine (DEN) and carbon tetrachloride (CCl4) in wild-type and LGR4 deficient mice, the effect of LGR4 on HCC was examined by histopathological evaluation and biochemical assays. Results: LGR4 expression was up-regulated in HCC samples, and its expression level was positively correlated with tumor size, microvascular invasion (MVI), TNM stage and pathological differentiation grade of HCC patients. In the mouse HCC model induced by DEN+CCl4, knockdown of LGR4 effectively inhibited the progression of HCC. Silencing of LGR4 inhibited the proliferation, migration, invasion, stem cell-like properties and Warburg effect of HCC cells. These phenotypes were promoted by R-spondin2 (Rspo2), an endogenous ligand for LGR4. Rspo2 markedly increased the nuclear translocation of β-catenin, whereas IWR-1, an inhibitor of Wnt/β-catenin signaling, reversed its effect. Deficiency of LGR4 significantly reduced the nuclear translocation of β-catenin and the expression of its downstream target genes cyclinD1 and c-Myc. Conclusions: LGR4 promotes HCC progression via Wnt/β-catenin signaling pathway.

F-box and WD repeat domain containing 7 (FBXW7) is an aboriginal and high-frequency mutant gene associated with esophageal squamous cell carcinoma (ESCC). This study was designed to determine the clinical value and molecular mechanisms of FBXW7 in the development of ESCC. The clinical significance of FBXW7 was analyzed in ESCC from TCGA data. The effects of FBXW7 on proliferation, colony formation, migration and invasion, angiogenesis, and apoptosis were tested in ESCC cells. PCR-array, sphere formation assay, and quantitative real-time polymerase chain reaction (qPCR) were used to explore the mechanism of FBXW7. FBXW7 was a significantly mutated gene in ESCC. It was an independent and potential predictor for survival in ESCC patients. In addition, FBXW7 overexpression significantly inhibited ESCC cell proliferation, migration, invasion, angiogenesis, and promoted cell apoptosis. PCR array revealed that FBXW7 overexpression leads to a significant change of gene expressions associated with angiogenesis, cell senescence, and DNA damage and repair. Sphere formation assay and qPCR showed FBXW7 was associated with ESCC stem cell formation. Our results suggest that FBXW7 may act as a tumor suppressor by repressing cancer stem cell formation and regulating tumor angiogenesis, cell senescence, DNA damage, and repair in ESCC.

Single-nucleotide polymorphisms in G protein-coupled receptor 180 (GPR180) are associated with hypertriglyceridemia. The aim of this study was to determine whether hepatic GPR180 impacts lipid metabolism. Hepatic GPR180 was knocked down using two approaches: Gpr180-specific short hairpin (sh)RNA carried by adeno-associated virus 9 (AAV9) and alb-Gpr180(-/-) transgene established by crossbreeding albumin-Cre mice with Gpr180(flox/flox) animals, in which Gpr180 was specifically knocked down in hepatocytes. Adiposity, hepatic lipid contents, and proteins related to lipid metabolism were analyzed. The effects of GPR180 on triglyceride and cholesterol synthesis were further verified by knocking down or overexpressing Gpr180 in Hepa1-6 cells. Gpr180 mRNA was upregulated in the liver of HFD-induced obese mice. Deficiency of Gpr180 decreased triglyceride and cholesterol contents in the liver and plasma, ameliorated hepatic lipid deposition in HFD-induced obese mice, increased energy metabolism, and reduced adiposity. These alterations were associated with downregulation of transcription factors SREBP1 and SREBP2, and their target acetyl-CoA carboxylase. In Hepa1-6 cells, Gpr180 knockdown decreased intracellular triglyceride and cholesterol contents, whereas its overexpression increased their levels. Overexpression of Gpr180 significantly reduced the PKA-mediated phosphorylation of substrates and consequent CREB activity. Hence, GPR180 might represent a novel drug target for intervention of adiposity and liver steatosis.

随着检测技术的进步与生物信息学的发展，人类不断从健康者或非菌血症疾病患者外周血中检出细菌或细菌成分，并对其生物学功能进行了初步探究，发现循环细菌的多样性、组成及各菌相对丰度的改变均可能是特定疾病发生与进展的重要驱动力，由此开启了学术界对循环细菌的新认识和讨论。现总结有关研究证据，阐述循环细菌在健康维持或非菌血症疾病发病机制中的角色，并进一步指明该主题中亟待回答的关键科学问题，以推动靶向循环细菌干预策略的开发与临床应用。

由诱生和效应2个阶段构成的干扰素应答是抗病毒与肿瘤免疫的经典生物学事件，但干扰素拮抗与药物毒性限制了干扰素的临床应用。新近大量实验证据揭示了肠道菌群及其代谢产物与干扰素应答网络在病毒与肿瘤性疾病中的关键作用，为精准修正肠道菌群实现抗病毒或肿瘤治疗带来新的思路。现就肠道菌群调控干扰素应答在病毒与肿瘤性疾病中的作用进行概述，以期为进一步基础和临床转化研究提供参考依据。

Abstract Background Esophageal cancer is a gastrointestinal tumor with high morbidity and mortality worldwide. FBXW7, is an aboriginal and high frequency mutant gene associated with ESCC. However, the current understanding of its clinical significance and mechanism in ESCC is not comprehensive. Methods Our previous data from WGS / WES and TCGA databases were used to analyze the clinical significance of FBXW7 in ESCC. Gene function and PCR-array were performed to explore the potential mechanism of FBXW7 in ESCC. Results The clinical information analysis revealed that low expression of FBXW7 is associated with poor prognosis in ESCC patients. Especially in those age≤55 years old, without drinking history and T3 stage, low expression of FBXW7was associated with poor prognosis. In addition, we found that overexpression of FBXW7 inhibited cell proliferation, migration, invasion and angiogenesis, and promoted cell apoptosis. PCR-array results showed that overexpression of FBXW7 resulted in a variable spectrum of tumor-associated gene expression in esophageal squamous cell carcinoma cells. Significant changes in gene expression related to angiogenesis, DNA damage repair, and cell senescence were observed. The changes of these pathway genes may be related to the regulation of FBXW7 on the stemness of ESCC tumor cells. Conclusions Our study investigated a novel role and mechanism of FBXW7 in esophageal squamous carcinoma and opened up new ideas for the clinical treatment of esophageal squamous carcinoma.

Aims: Esophageal squamous cell carcinoma (ESCC) is one of the most prevalent malignancies with unfavorable clinical outcomes and limited therapeutic methods. As a key enzyme in RNA metabolism, debranching RNA Lariats 1 (DBR1) is involved in intron turnover and biogenesis of noncoding RNA. Although cancer cells often show disorder of nucleic acid metabolism, it is unclear whether DBR1 has any effect on the carcinogenesis and progression of ESCC. Methods: Here we detected DBR1 expression in 112 ESCC samples by immunohistochemistry and analyzed its correlation with clinical parameters and survival. Results: DBR1 is mainly located in the nucleus of ESCC tissue. And DBR1 was associated with several malignant clinical features in patients, including tumor location ( χ 2 = 9.687, P = .021), pathologic T stage ( χ 2 = 5.771, P = .016), lymph node metastasis ( χ 2 = 8.215, P = .004) and N classification ( χ 2 = 10.066, P = .018). Moreover, Kaplan-Meier analysis showed that ESCC patients harboring lower DBR1 expression had a worse prognosis in comparison with those with higher DBR1 expression ( P = .005). Univariate and multivariate Cox proportional hazards regression analyses indicated that decreased DBR1 might act as an independent predictor of poor prognosis for ESCC patients. Conclusion: Abnormal RNA metabolism might play a critical role in promoting the progression of ESCC, and DBR1 may be a promising potential biomarker for predicting the prognosis of ESCC patients.

Esophageal squamous cell carcinoma (ESCC) demonstrates high genome instability. Here, we analyze 528 whole genomes to investigate structural variations’ mechanisms and biological functions. SVs show multi-mode distributions in size, indicating distinct mutational processes. We develop a tool and define five types of complex rearrangements with templated insertions. We highlight a type of fold-back inversion, which is associated with poor outcomes. Distinct rearrangement signatures demonstrate variable genomic metrics such as replicating time, spatial proximity, and chromatin accessibility. Specifically, fold-back inversion tends to occur near the centrosome; TD-c2 (Tandem duplication-cluster2) is significantly enriched in chromatin-accessibility and early-replication region compared to other signatures. Analyses of TD-c2 signature reveal 9 TD hotspots, of which we identify a hotspot consisting of a super-enhancer of PTHLH . We confirm the oncogenic effect of the PTHLH gene and its interaction with enhancers through functional experiments. Finally, extrachromosomal circular DNAs (ecDNAs) are present in 14% of ESCCs and have strong selective advantages to driver genes.

Aims: Esophageal squamous cell carcinoma (ESCC) is one of the most prevalent malignancies with unfavorable clinical outcomes and limited therapeutic methods. As a key enzyme in RNA metabolism, debranching RNA Lariats 1 (DBR1) is involved in intron turnover and biogenesis of noncoding RNA. Although cancer cells often show disorder of nucleic acid metabolism, it is unclear whether DBR1 has any effect on the carcinogenesis and progression of ESCC. Methods: Here we detected DBR1 expression in 112 ESCC samples by immunohistochemistry and analyzed its correlation with clinical parameters and survival. Results: DBR1 is mainly located in the nucleus of ESCC tissue. And DBR1 was associated with several malignant clinical features in patients, including tumor location ( χ 2 = 9.687, P = .021), pathologic T stage ( χ 2 = 5.771, P = .016), lymph node metastasis ( χ 2 = 8.215, P = .004) and N classification ( χ 2 = 10.066, P = .018). Moreover, Kaplan-Meier analysis showed that ESCC patients harboring lower DBR1 expression had a worse prognosis in comparison with those with higher DBR1 expression ( P = .005). Univariate and multivariate Cox proportional hazards regression analyses indicated that decreased DBR1 might act as an independent predictor of poor prognosis for ESCC patients. Conclusion: Abnormal RNA metabolism might play a critical role in promoting the progression of ESCC, and DBR1 may be a promising potential biomarker for predicting the prognosis of ESCC patients. Keywords DBR1 , ESCC , prognosis , biomarker

BACKGROUND:CDCA7 is a copy number amplified gene identified not only in esophageal squamous cell carcinoma (ESCC) but also in various cancer types. Its clinical relevance and underlying mechanisms in ESCC have remained unknown.METHODS:Tissue microarray data was used to analyze its expression in 179 ESCC samples. The effects of CDCA7 on proliferation, colony formation, and cell cycle were tested in ESCC cells. Real-time PCR and Western blot were used to detect the expression of its target genes. Correlation of CDCA7 with its target genes in ESCC and various SCC types was analyzed using GSE53625 and TCGA data. The mechanism of CDCA7 was studied by chromatin immunoprecipitation (ChIP), luciferase reporter assays, and rescue assay.RESULTS:The overexpression of CDCA7 promoted proliferation, colony formation, and cell cycle in ESCC cells. CDCA7 affected the expression of cyclins in different cell phases. GSE53625 and TCGA data showed CCNA2 expression was positively correlated with CDCA7. The knockdown of CCNA2 reversed the malignant phenotype induced by CDCA7 overexpression. Furthermore, CDCA7 was found to directly bind to CCNA2, thus promoting its expression.CONCLUSIONS:Our results reveal a novel mechanism of CDCA7 that it may act as an oncogene by directly upregulating CCNA2 to facilitate tumor progression in ESCC.

Asprosin is a new hormone released from white adipose tissue (WAT) that not only promotes glucose release in the liver but also activates orexigenic neurons in the hypothalamus to promote appetite and weight gain. Its effect on skeletal muscle glucose uptake is unclear. This research, a stable asprosin expression system was formed by first constructing a eukaryotic expression vector pPIC9K-8His-Asprosin, and then transforming it into the Pichia pastoris strain GS115. Pichia pastoris methanol induction combined with Nickel-NTA magnetic beads purification strategy was used to express and purify asprosin protein. Purified asprosin can promote the phosphorylation of PKA substrate, and intraperitoneal injection of asprosin can increase blood glucose. After proteolysis and detection by mass spectrometry, asprosin was found to have 3 glycosylation sites and multiple glycosyl types. Asprosin up-regulated glucose transporter 4 (GLUT4) expression in myotubes, including mRNA and protein levels. In addition, asprosin enhanced AMP-activated protein kinase (AMPK) phosphorylation, but it had no effect on AKT phosphorylation with or without insulin treatment. Treatment with an AMPK inhibitor (compound C) reduced the asprosin-mediated glucose uptake effect. These results show that purified asprosin activated AMPK signaling in skeletal muscle and further promoted glucose uptake. From the perspective of skeletal muscle uptake of glucose, asprosin may have beneficial effects on type 2 diabetes.