Glioblastoma is the most aggressive primary brain tumor in adults, with limited therapeutic success and, therefore, poor prognosis. Its malignancy is partly driven by the high proliferative capacity of glioblastoma cells, yet the underlying molecular mechanisms remain unclear. Recent studies have revealed transcriptomic similarities between glioblastoma cells and human fetal neural stem/progenitor cells (NSCs), suggesting that glioblastoma may exploit developmental programs that promote NSC proliferation. Fetal human NSCs rely on glutaminolysis-a metabolic pathway induced by the human-specific mitochondrial protein ARHGAP11B-to sustain proliferation. Here, we show that ARHGAP11B expression correlates with glioma malignancy and is essential for glioblastoma cell proliferation, implicating a critical role of glutaminolysis in tumor growth. Among glutaminolysis-related enzymes, glutamic-oxaloacetic transaminase 2 (GOT2) shows a strong positive correlation with glioma grade and poor patient prognosis. Functional assays reveal that GOT2 knockdown significantly suppresses glioblastoma cell growth, indicating that GOT2-mediated glutaminolysis is critical for their proliferation. Metabolomic profiling further shows that GOT2 is required for nucleotide precursor synthesis, underscoring its role in supporting DNA replication. Consistently, GOT2 depletion reduces the proportion of glioblastoma cells in the S phase of the cell cycle. These findings suggest glioblastoma cells hijack an evolutionarily adapted metabolic program to support malignant growth.

Glucose deprivation is a major metabolic stress that requires coordinated adaptive responses to maintain cellular homeostasis and survival, yet the role of tripartite motif-containing 24 (TRIM24) in this process remains unclear. To address this question, we generated CRISPR-Cas9-mediated TRIM24-knockout MCF-7 and HEK293 cell lines, performed targeted metabolomic profiling and aspartate assays, used 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR), aminooxyacetic acid (AOA), aspartate supplementation, and glutamic-oxaloacetic transaminase 2 (GOT2) knockdown to probe AMPK signaling and aspartate metabolism, and examined starvation responses in constitutive Trim24 knockout mice on a C57BL/6 background. Loss of TRIM24 sensitized cells to glucose deprivation. Re-expression of TRIM24 partially restored cell viability under glucose deprivation in both MCF-7 and HEK293 cells. Under glucose-free conditions, TRIM24 deficiency was associated with impaired AMP-activated protein kinase (AMPK) pathway activation, increased intracellular aspartate accumulation, and altered ATP/AMP levels. Pharmacological reactivation of AMPK by AICAR improved the survival of TRIM24-deficient cells under glucose deprivation. Reducing intracellular aspartate by AOA treatment or GOT2 knockdown restored AMPK pathway activation and improved adaptation to glucose deprivation, whereas exogenous aspartate suppressed AMPK signaling and increased ATP/AMP levels. In vivo, starvation of Trim24-deficient mice was associated with reduced AMPK pathway activation and increased aspartate levels. Together, these findings support a model in which TRIM24 contributes to adaptation to glucose deprivation and in which abnormal aspartate accumulation contributes to impaired AMPK pathway activation in TRIM24-deficient cells.

Hepatocellular carcinoma (HCC), the most common subtype of primary liver cancer, is characterized by a poor prognosis, with most patients diagnosed at advanced stages that are unresectable and resistant to targeted therapies. This underscores the critical need for novel therapeutic targets. In this study, we investigated the role of glutamate-oxaloacetate transaminase 1 (GOT1) in HCC progression and evaluated its therapeutic potential. GOT1 expression was markedly downregulated in HCC tissues compared with adjacent non-tumor tissues, and higher GOT1 expression correlated with improved patient survival. Functional experiments revealed that overexpression of GOT1 suppressed HCC cell proliferation and tumor growth while promoting apoptosis both in vitro and in vivo. Mechanistically, integrated transcriptome sequencing and mass spectrometry identified solute carrier family 25 member 5 (SLC25A5) as a GOT1-interacting partner. GOT1-induced oxidative stress, mitochondrial membrane depolarization, and activation of the apoptotic cascade were mediated through upregulation of SLC25A5; Conversely, SLC25A5 knockdown rescued these phenotypic effects. Importantly, adeno-associated virus-mediated delivery of GOT1 (AAV-TBG-GOT1) significantly inhibited tumor growth in preclinical HCC models, underscoring its translational relevance. Collectively, these results establish GOT1 as a tumor suppressor in HCC that acts via SLC25A5-dependent mitochondrial apoptosis and propose GOT1 as a promising prognostic indicator and therapeutic target for HCC.

IntroductionHyperlipidemia remains a significant risk factor for cardiovascular diseases and is a leading cause of death, driving the need for novel and effective cholesterol-lowering agents. Preussin (1) has recently demonstrated lipid-lowering effects by inhibiting intestinal cholesterol absorption in human colorectal adenocarcinoma (Caco-2) cells and in an ex vivo intestinal loop in rats, comparable to those of ezetimibe. Ezetimibe is the only drug that targets reducing intestinal cholesterol absorption. Recently, two natural (preussin C, 2, and preussin B, 4) and three unnatural (3, 5, and 6) analogues of preussin have been synthesized and also displayed interesting lipid-lowering effects in human hepatocellular carcinoma (HepG2) cells. However, the underlying mechanisms and the potential lipid-lowering effects of preussin and its synthetic analogues in inhibiting cholesterol absorption are not yet fully understood.MethodsThis study aims to evaluate the inhibitory effect of cholesterol absorption by preussin and its analogues using a fluorescent-micelle cholesterol transport in intestinal Caco-2 cells, which further confirmed by an in vivo cholesterol absorption assay. The most potent analogue was further investigated for its cellular and molecular mechanisms in reducing lipid levels and identifying possible target proteins.ResultsAll synthetic derivatives markedly inhibited cholesterol absorption in the intestinal Caco-2 cells to a similar extent as preussin and ezetimibe. However, only compound 4 (preussin B) displayed the most significant reduction in plasma cholesterol, identical to preussin and ezetimibe, with similar potency in rats. The precise mechanisms and potential targets of this potent compound were additionally identified using protein binding assay and label-free quantification via proteomics analysis. The results revealed substantial differential expression in four proteins associated with lipid metabolism. Notably, glutamic-oxaloacetic transaminase 2 (GOT2), one of the altered proteins, was shown to interact with compound 4 in a protein binding assay. Molecular dynamics simulation analysis indicated that compound 4 binds to a pocket on GOT2 comparable to that of its natural cofactor. This interaction, combined with the observed downregulation of GOT2 expression, contributed to the inhibition of cholesterol absorption.ConclusionThese findings suggest that synthetic compound 4 (preussin B) is a promising candidate for inhibiting cholesterol absorption in the treatment of hyperlipidemia.

Salvianolic acid A targets glutamic-oxaloacetic transaminase 2 to ameliorate doxorubicin-induced myocardial oxidative injury by activating malate-aspartate NADH shuttle.

Colorectal cancer (CRC) is a global health issue requiring novel diagnostic and therapeutic approaches to improve patient outcomes. Glutamate oxaloacetate transaminase 1 (GOT1) plays a crucial role in metabolism and is associated with various cancers. However, its expression and potential as a diagnostic marker in CRC have not been thoroughly investigated. We analysed The Cancer Genome Atlas data on GOT1 normalised to transcripts per million. We used tools such as Gene Expression Profiling Interactive Analysis 2, logistic regression, receiver operating characteristic analysis, Sieber algorithm for immune detection, immune checkpoint gene correlation, cBioPortal analysis, and genomic sensitivities of cancers from the Genomics of Drug Sensitivity in Cancer and Cancer Therapeutics Response Portal to evaluate the association of GOT1 with CRC. GOT1 expression significantly varied across cancer types, showing high diagnostic value in colon adenocarcinoma and rectal adenocarcinoma. Moreover, it correlated with immune cells, such as CD8+ T and plasma cells, and immune checkpoint genes LGALS9 and TNFRSF4. Tumour genetic variations differed in mutation burden, copy number alterations, and microsatellite instability. Drug sensitivities, including those of navitoclax and CCT036477, showed an association with GOT1 expression. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested the involvement of GOT in cellular metabolism. Our comprehensive analysis revealed a critical role of GOT1 in CRC, confirming its role as a diagnostic and therapeutic target. Nonetheless, its role in tumorigenesis warrants further investigation.

ABSTRACTOxaloacetate (OAA) is converted to aspartate by mitochondrial glutamic‐oxaloacetic transaminase 2 (GOT2) along with the conversion of glutamate to alpha‐ketoglutarate (α‐KG). Glutamate can also be directly converted to α‐KG by glutamate dehydrogenase. In past work, we found that in skeletal muscle mitochondria energized by succinate alone, oxaloacetate accumulates and inhibits succinate dehydrogenase (complex II) in a manner dependent on inner membrane potential (ΔΨ). Here, we tested the hypothesis that deleting GOT2 would increase OAA concentrations, decrease complex II‐energized respiration, and alter the selectivity of succinate versus glutamate for energy. Incubating wild‐type mitochondria with succinate and glutamate revealed that increments in ADP increased OAA and caused a preferential use of glutamate for energy. Deletion of GOT2 compared to wild‐type decreased complex II energized respiration, increased OAA, and decreased consumption of glutamate relative to succinate. OAA accumulation was also associated with decreased conversion of succinate to fumarate and malate. These findings are consistent with GOT2 control of metabolite flow through succinate dehydrogenase via regulation of OAA and consequent inhibition of succinate dehydrogenase. In contrast to respiration energized at complex II, when mitochondria were energized at complex I by pyruvate + malate, respiration did not differ between GOT2KO and WT mitochondria, and oxaloacetate was not detectable. In summary, GOT2 and OAA mediate complex II respiration and mitochondrial energy substrate selectivity.

Glutamine has emerged as a focus of cancer metabolism research, although its role in liver hepatocellular carcinoma (LIHC) has yet to be fully elucidated. To determine the role of glutamine metabolism in the development of LIHC, the gene expression profiles and the clinical data of patients with LIHC were obtained from The Cancer Genome Atlas database and the International Cancer Genome Consortium website. Consensus clustering was used to identify distinct molecular clusters. Functional en 10.3892/ol.2025.15149 richment analysis between clusters was performed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases, and gene set variation analysis was performed. Least absolute shrinkage and selection operator and multivariate Cox regression analyses were then performed to generate a novel prognostic model. The prognostic, immune, mutational and drug-sensitive characteristics of the model were subsequently evaluated. The clinical proteomic tumor analysis consortium and reverse transcription-quantitative PCR analysis were then used to assess the protein and mRNA expression levels of the modeled genes. In addition, western blot analysis and Cell Counting Kit-8, 5-ethynyl-2′-deoxyuridine, Transwell and wound healing assays were performed to further evaluate the role of glutamate-oxaloacetate transaminase 2 (GOT2) in the pathogenesis of LIHC. Data from multiple LIHC cohorts were utilized to identify two distinct clusters of LIHC, each characterized by unique clinical and immunological features associated with different levels of glutamine metabolism-related genes. Numerous functional pathway differences were identified between these clusters, and these were demonstrated to be crucial for the onset and progression of LIHC. For modeling of glutamine metabolism-related features, patients with LIHC were divided into two groups, namely a high- and a low-risk group. Different clusters of patients with LIHC exhibited distinct characteristics in terms of their clinicopathological features, drug-sensitivity and mutations. For example, the high-risk group had a higher mutational load and was associated with a poorer prognosis compared to the low-risk group. Finally, GOT2 protein and mRNA expression levels were significantly lower in LIHC tissues compared to paracancerous tissues, and GOT2 knockdown promoted the malignant phenotype of LIHC. In conclusion, the results of the present study indicate that glutamine metabolism exerts a crucial role in the tumorigenesis and progression of LIHC, and that this is positively associated with poor prognosis. The identified glutamine metabolism-related signature was revealed to have notable accuracy in predicting the prognosis and immune characteristics of patients with LIHC. Moreover, the expression level of GOT2 was downregulated in LIHC, and a low expression of GOT2 was indicative of a poor prognosis for patients with LIHC, suggesting that the expression of GOT2 may be used as a potential therapeutic target.

Glutamate-oxaloacetate transaminase 2 (GOT2) has been demonstrated to contribute to lung cancer cell growth, invasion, migration and angiogenesis. Herein, we further probed the functions of GOT2 on lung adenocarcinoma (LUAD) cell ferroptosis and immune escape and its associated mechanism. qRT-PCR and Western blot analysis analyses were used to detect the levels of GOT2, and Yin Yang 1 (YY1). A mouse xenograft model was established for in vivo analysis. CCK-8, 5-ethynyl-2'-deoxyuridine, and wound healing assays were applied for the detection of cell proliferation and migration. Cell ferroptosis was evaluated by flow cytometry and the levels of malondialdehyde (MDA) and Fe2+. Immune escape was assessed by measuring CD8+ T cell apoptosis and programmed death-1 ligand 1 (PD-L1) levels. The interaction between GOT2 and YY1 was determined using Chromatin immunoprecipitation and dual luciferase reporter assays. GOT2 expression was higher in LUAD tissues and cells, and the silencing of GOT2 impeded LUAD growth in vivo. Further loss-of-function assays showed that GOT2 silencing suppressed LUAD cell proliferation, migration and immune escape, and induced ferroptosis. Mechanically, we found that YY1 activated the transcription of GOT2 and could elevate GOT2 expression. Moreover, YY1 silencing repressed LUAD cell proliferation, migration and immune escape, and evoked ferroptosis, while theses effects could be reversed by GOT2 overexpression. YY1 activated GOT2 and elevated the expression of GOT2, which then promoted LUAD cell growth, migration and immune escape, and suppressed cell ferroptosis, suggesting a novel perceptivity for the treatment of LUAD.

Uncoupling protein 3 protects against pathological cardiac hypertrophy via downregulation of aspartate.

The domesticated silkworm (Bombyx mori) has evolved a highly efficient nitrogen utilization system to support silk production. The silk glands play a pleiotropic role in sequestering nitrogen resources for silk synthesis, mitigating aminoacidemia by assimilating free amino acids, and reallocating nitrogen during metamorphosis through programmed cell death. However, the specific functions of nitrogen metabolism-related genes in this process remain unclear. Using CRISPR/Cas9-based gene editing, mutations were generated in glutamine synthetase (GS), glutamate synthetase (GOGAT), asparagine synthetase (AS), glutamate dehydrogenase (GDH) and glutamate oxaloacetate transaminase 1 (GOT1). Disruption of GS, GOGAT, and AS consistently reduced silkworm cocoon and pupal weight and significantly down-regulated silk protein gene transcription, whereas GOT1 mutation had no such effect. GOGAT mutants exhibited abnormally enlarged silk glands, whereas GS and AS mutants showed delayed programmed cell death in the silk glands. In contrast, GOT1 mutants displayed normal silk gland morphology but were consistently smaller. Disruption of GS, GOGAT, and AS led to more extensive transcriptional changes, including altered expression of transcription factors in the silk glands, compared with GOT1 mutants. Both GS and GOGAT mutants exhibited up-regulation of AS and GDH, while only GOGAT mutants displayed elevated AS enzymatic activity, suggesting that GOGAT may compete with AS for glutamine in the silk glands to support silk protein synthesis. AS mutants showed significantly elevated GOT activity and up-regulation of several metabolic pathways, indicating that AS may functionally interact with GOT in regulating both silk gland development and programmed cell death during metamorphosis.

Mitochondrial aspartate aminotransferase (GOT2) protein as a potential cryodamage biomarker in rooster spermatozoa cryopreservation.

METTL16 controls airway inflammations in smoking-induced COPD via regulating glutamine metabolism.

TPS2681 Background: Glypican-3 (GPC3) is a membrane-bound heparin sulfate proteoglycan involved in cell proliferation that is overexpressed in several tumor types. BOXR1030 is an autologous T-cell therapy that co-expresses a chimeric antigen receptor (CAR) targeting GPC3 on tumor cells and glutamic-oxaloacetic transaminase 2 (GOT2), a mitochondrial enzyme that plays an important role in maintaining mitochondrial function and improving T-cell functionality in the solid tumor microenvironment. In non-clinical studies, BOXR1030 showed GPC3-specific cytotoxicity, proliferation and cytokine release only in the presence of GPC3+ cells. Methods: DUET-01 (NCT05120271) is an open-label, dose escalation and expansion clinical trial to determine a safe dose of BOXR1030 in patients with advanced GPC3-positive solid tumors. Dose escalation will be guided by the occurrence of dose-limiting toxicities (DLTs) within 28 days of dosing. The Bayesian logistic regression model will be used on the accumulated DLT data to estimate the maximum tolerated dose (MTD). The Dose Escalation Committee will select the recommended phase 2 dose (RP2D) to be used in the expansion phase cohort(s) (10-20 subjects each) based on data accumulated in the dose escalation cohorts. Eligible subjects will undergo leukapheresis to obtain T cells for BOXR1030 manufacturing and will receive 3 days of lymphodepleting chemotherapy with fludarabine and cyclophosphamide within 5 days prior to BOXR1030 administration. The starting dose is 0.3 × 106 BOXR1030 T cells/kg body weight. Antitumor activity will be assessed every 6 weeks after cell infusion per Response Evaluation Criteria In Solid Tumors (RECIST) version 1.1 and RECIST for immune-based therapeutics (iRECIST). Six months after BOXR1030 administration, subjects will enter long-term follow-up for up to 15 years. Long-term follow-up assessments will focus on long-term safety and disease status. Main inclusion criteria are histologically confirmed advanced unresectable or metastatic hepatocellular carcinoma, squamous cell carcinoma of the lung, myxoid/round cell liposarcoma, or Merkel cell carcinoma; GPC3 overexpression by immunohistochemistry assay confirmed centrally on tumor specimen taken within 6 months prior to signing consent and after the initiation of the subject’s most recent systemic anticancer therapy; body weight of ≥ 50 kg (≥ 65 kg for dose level 1); and life expectancy > 16 weeks. The primary endpoints are the incidence of DLTs; determination of the MTD and RP2D; the type, frequency and severity of treatment-emergent adverse events; clinically significant abnormal safety laboratory findings and vital signs. Key secondary endpoints include efficacy parameters such as objective response rate, BOXR1030 T-cell expansion and persistence, and levels of inflammatory markers and cytokines. The first patient was treated with BOXR1030 in December 2022. Clinical trial information: NCT05120271 .

Abstract At present, pancreatic cancer stands as the 3rd cause of cancer-related fatalities in the US. The most common type of pancreatic cancer develops from the exocrine pancreas known as ductal adenocarcinoma (PDAC). PDAC exhibits a highly aggressive nature and carries a bleak prognosis, with a 5-year survival rate averaging below 10%. Moreover, the majority of patients receive their diagnosis at advanced stages of the disease. Compounding the issue, the emergence of resistance to modern first-line chemotherapies contributes to the progression of PDAC and exacerbates its overall high mortality rate. The specific cause of PDAC remains unknown, although lifestyle factors such as alcohol and tobacco use, along with genetic abnormalities, have been linked to its increasing incidence over the past decade. Mutations in KRAS, TP53, SMAD4, and CDKN2A are recognized as genetic drivers contributing to the progression of PDAC. Recently, research has shifted its focus to investigating the role of epigenetic dysregulation in PDAC advancement. Significant evidence suggests that remodeling of the epigenome plays a crucial role in the development and progression of PDAC, but the exact mediators responsible for these alterations remain largely unknown. Recent studies have indicated that mitochondrial enzymes may influence epigenetic modifications, thus contributing to the progression of tumorigenesis. In our previous work, we reported that the mitochondrial protein GOT2 (glutamic-oxaloacetic transaminase) supports PDAC development by promoting an immunosuppressive phenotype within the tumor microenvironment. Ongoing epigenetic studies following ChIP- and ATAC-seq (chromatin immunoprecipitation and assay for transposase-accessible chromatin with sequencing, respectively) experiments in mammalian PDAC cell lines have revealed that loss of GOT2 leads to genome-wide changes in histone acetylation and chromatin accessibility. These changes in acetylation and chromatin architecture imply a reconfiguration of the epigenetic landscape and subsequent alterations in the PDAC transcriptome, suggesting a potential role for GOT2 in maintaining epigenetic profiles within the context of pancreatic cancer. Further exploration of the genomic regions where these histone modification changes occur has implicated cis-regulatory regions and the expression of their target genes, which may be crucial for PDAC progression. Collectively, these epigenetic studies hold the potential to identify novel therapeutic targets that could address the urgent need for more effective antineoplastic treatments against this aggressive form of pancreatic cancer. Citation Format: Luis Francisco Diaz, Mara H. Sherman, Jonathan R. Brody. A link between the mitochondrial enzyme glutamic-oxaloacetic transaminase 2 (GOT2) and epigenetic dysregulation in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr A073.

Prostate cancer (PCa) poses a significant health threat to males, and research has shown that fish oil (FO) can impede PCa progression by activating multiple mitochondria-related pathways. Our research is focused on investigating the impact of FO on succinylation, a posttranslational modification that is closely associated with mitochondria in PCa cells. This study employed a mass spectrometry-based approach to investigate succinylation in PCa cells. Bioinformatics analysis of these succinylated proteins identified glutamic-oxaloacetic transaminase 2 (GOT2) protein as a key player in PCa cell proliferation. Immunoprecipitation and RNA interference technologies validated the functional data. Further analyses revealed the significance of GOT2 protein in regulating nucleotide synthesis by providing aspartate, which is critical for the survival and proliferation of PCa cells. Our findings suggest that FO-dependent GOT2 succinylation status has the potential to inhibit building block generation. This study lays a solid foundation for future research into the role of succinylation in various biological processes. This study highlights the potential use of FO as a nutrition supplement for managing and slowing down PCa progression.

Identification and characterization of tryptophan-kynurenine pathway-related genes involving lamprey (Lampetra japonica) innate immunity.

System analysis based on glutamine catabolic-related enzymes identifies GPT2 as a novel immunotherapy target for lung adenocarcinoma

Malate-aspartate shuttle (MAS) is essential for maintaining glycolysis and energy metabolism in tumors, while its regulatory mechanisms in neuroblastoma (NB), the commonest extracranial malignancy during childhood, still remain to be elucidated. Herein, by analyzing multi-omics data, GATA binding protein 2 (GATA2) and its antisense RNA 1 (GATA2-AS1) were identified to suppress MAS during NB progression. Mechanistic studies revealed that GATA2 inhibited the transcription of glutamic-oxaloacetic transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2). As a long non-coding RNA destabilized by RNA binding motif protein 15-mediated N6-methyladenosine methylation, GATA2-AS1 bound with far upstream element binding protein 3 (FUBP3) to repress its liquid-liquid phase separation and interaction with suppressor of zest 12 (SUZ12), resulting in decrease of SUZ12 activity and epigenetic up-regulation of GATA2 and other tumor suppressors. Rescue experiments revealed that GATA2-AS1 inhibited MAS and NB progression via repressing interaction between FUBP3 and SUZ12. Pre-clinically, administration of lentivirus carrying GATA2-AS1 suppressed MAS, aerobic glycolysis, and aggressive behaviors of NB xenografts. Notably, low GATA2-AS1 or GATA2 expression and high FUBP3, SUZ12, GOT2 or MDH2 levels were linked with unfavorable outcome of NB patients. These findings suggest that GATA2-AS1 inhibits FUBP3 phase separation to repress MAS and NB progression via modulating SUZ12 activity.

Glutamine metabolism is critical for development of hepatocellular carcinoma (HCC), which makes it a novel promising treatment target. However, clinical evidence suggested glutamine withdrawal therapy does not achieved the desired tumor suppression. Therefore, it is valuable to investigate the survival mechanisms of tumors with glutamine deprivation. The HCC cells were cultured in glutamine-free medium or supplemented with glutamine metabolites or ferroptosis inhibitors. The parameters related to ferroptosis and the activity of GSH synthesis-related enzymes of the HCC cells were detected by corresponding kits. The expressions of glutamate oxaloacetate transaminase 1 (GOT1), c-Myc and Nrf2 were detected by western blot and qRT-PCR. The chromatin immunoprecipitation and luciferase reporter assays were performed to investigate the correlation between c-Myc and GOT1. The siRNAs of c-Myc and GOT1 were used to explore their roles in GSH (GSH) synthesis and ferroptosis in vitro and in vivo. Glutamine deprivation-induced ferroptosis did not completely inhibit HCC cellsproliferation. Glutamine deprivation activated the expression of c-Myc, which promoted the transcription of GOT1 and Nrf2, consequently maintaining the GSH synthesisand inhibiting ferroptosis. In addition, combined inhibition of GOT1 with glutamine deprivation could result in better inhibition of HCC in vitro and in vivo. In our work, the results indicate that GOT1 induced by c-Myc may play an important role in combating ferroptosis due to glutamine deprivation, making it a significant target in glutamine withdrawal therapy. This study provides a theoretical foundation for the clinical targeted therapy for HCC.