A novel cuproptosis-related gene signature for the prediction of liver cancer prognosis identified DLAT is a potential therapeutic target

Background and Purpose The liver cancer (LC) is a highly malignant tumor of the digestive system with a poor prognosis. Cuproptosis is a new type of regulated cell death that has been found by researchers. The expression of cuproptosis-related genes in LC and their relevance to prognosis, on the other hand, remain unknown. This study aimed to explore a gene signature to predict the liver cancer prognosis and identified the vital gene. Experimental approach The expression patterns of RNA and related clinical data of 371 LC patients were obtained based on The Cancer Genome Atlas (TCGA). Differentially expressed genes (DEGs) were acquired by comparing tumors with adjacent normal samples. Genes displaying significant association with OS were screened through univariate Cox regression analysis and the least absolute shrinkage and selection operator (LASSO) algorithm. All cases were classified into the validation or training group to validate the constructed gene signature. We conducted real-time polymerase chain reaction (PCR) and assays for transwell invasion, CCK-8, and colony formation to determine the biological roles of DLAT. Key Results The differential expression of twelve cuproptosis regulators in LC and normal liver tissues was discovered in this investigation. DEGs can be used to distinguish between two forms of LC. Cuproptosis-related genes were evaluated for survival predictive significance using the Cancer Genome Atlas (TCGA) cohort. A 3-gene signature based on least absolute shrinkage and selection operator (LASSO) Cox regression was used to categorize an LC patient cohort from the TCGA into low- and high-risk categories. Patients in the low-risk group had a considerably higher likelihood of surviving (P = 0.05) than those in the high-risk group. When paired with clinical parameters, risk score was an independent predictor in predicting the OS of patients with LC. Conclusions & Implications Cuproptosis-related genes thus play an important role in tumor formation and can be used to predict the prognosis of LC patients. DLAT has the best prognostic value and can be a therapeutic target for liver cancer. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement The Health and Family Planning Commission of Jiangsu Province [#Z2020069], the Health and Family Planning Commission of Nanjing, Jiangsu Province[#YKK20172], and the Inheriting Studio of National Famous Old Chinese Medicine Experts-Zhu Yongkang（National Traditional Chinese Medicine Science and Education 2022 No. 75）. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The study used (or will use) ONLY openly available human data that were originally located at TCGA database. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors. All data produced in the present work are contained in the manuscript. All data produced are available online at TCGA database. * LC : Liver cancer DEG : Differentially expressed genes TCGA : The Cancer Genome Atlas LASSO : Least absolute shrinkage and selection operator CCK8 : Cell counting kit OS : Overall survival SEER : Surveillance Epidemiology End Results LIPT1 : Lipolytransferase 1 LIAS : Lipoyl synthase DLD : Dihydrolipoamide dehydrogenase PDH : Pyruvate dehydrogenase ROS : Reactive oxygen species RNA-seq : RNA sequencing IC50 : Half maximal inhibitory concentration IHC : Immunohistochemistry IOD : Integrated optical density AOD : Average optical density ROC : Receiver operating characteristic curve AUC : Area under the ROC curve GO : Gene ontology KEGG : Kyoto Encyclopaedia of Genes and Genomes TCA : Tricarboxylic acid