{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T16:44:42Z","timestamp":1790181882037,"version":"4.1.0"},"reference-count":45,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2024,2,11]],"date-time":"2024-02-11T00:00:00Z","timestamp":1707609600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"publisher","award":["202106010223"],"award-info":[{"award-number":["202106010223"]}],"id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"publisher","award":["82172693"],"award-info":[{"award-number":["82172693"]}],"id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Natural Science Foundation of China","award":["202106010223"],"award-info":[{"award-number":["202106010223"]}]},{"name":"National Natural Science Foundation of China","award":["82172693"],"award-info":[{"award-number":["82172693"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hepatocellular carcinoma (HCC), the main pathological type of liver cancer, is related to risk factors such as viral hepatitis, alcohol intake, and non-alcoholic fatty liver disease (NAFLD). The constitutive activation of the PI3K\/AKT signaling pathway is common in HCC and has essential involvement in tumor progression. The serine\/threonine kinase AKT has several downstream substrates, which have been implicated in the regulation of cellular metabolism. However, the contribution of each of the three AKT isoforms, i.e., AKT1, AKT2 and AKT3, to HCC metabolism has not been comprehensively investigated. In this study, we analyzed the functional role of AKT1, AKT2 and AKT3 in HCC metabolism. The overexpression of activated AKT1, AKT2 and AKT3 isoforms in the human HCC cell lines Hep3B and Huh7 resulted in higher oxygen consumption rate (OCR), ATP production, maximal respiration and spare respiratory capacity in comparison to vector-transduced cells. Vice versa, lentiviral vector-mediated knockdowns of each AKT isoform reduced OCR in both cell lines. Reduced OCR rates observed in the three AKT isoform knockdowns were associated with reduced extracellular acidification rates (ECAR) and reduced lactate production in both analyzed cell lines. Mechanistically, the downregulation of OCR by AKT isoform knockdowns correlated with an increased phosphorylation of the pyruvate dehydrogenase on Ser232, which negatively regulates the activity of this crucial gatekeeper of mitochondrial respiration. In summary, our data indicate that each of the three AKT isoforms is able to upregulate OCR, ECAR and lactate production independently of each other in human HCC cells through the regulation of the pyruvate dehydrogenase.<\/jats:p>","DOI":"10.3390\/ijms25042168","type":"journal-article","created":{"date-parts":[[2024,2,12]],"date-time":"2024-02-12T04:47:45Z","timestamp":1707713265000},"page":"2168","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines"],"prefix":"10.3390","volume":"25","author":[{"given":"Ling-Yu","family":"Tian","sequence":"first","affiliation":[{"name":"Institute of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistra\u00dfe 52, 20246 Hamburg, Germany"},{"name":"Beijing Key Surgical Basic Research Laboratory of Liver Cirrhosis and Liver Cancer, Department of Hepatobiliary Surgery, Peking University People\u2019s Hospital, Beijing 100044, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3190-9511","authenticated-orcid":false,"given":"Daniel J.","family":"Smit","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistra\u00dfe 52, 20246 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadezhda V.","family":"Popova","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistra\u00dfe 52, 20246 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Horn","sequence":"additional","affiliation":[{"name":"Research Department Cell and Gene Therapy, Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lis Noelia","family":"Velasquez","sequence":"additional","affiliation":[{"name":"I. Department of Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany"},{"name":"Hamburg Center for Translational Immunology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9325-8227","authenticated-orcid":false,"given":"Samuel","family":"Huber","sequence":"additional","affiliation":[{"name":"I. Department of Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany"},{"name":"Hamburg Center for Translational Immunology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2176-6526","authenticated-orcid":false,"given":"Manfred","family":"J\u00fccker","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistra\u00dfe 52, 20246 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3430","DOI":"10.1002\/cncr.30237","article-title":"Comparison of hepatocellular carcinoma in Eastern versus Western populations","volume":"122","author":"Choo","year":"2016","journal-title":"Cancer"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1016\/S0140-6736(22)01200-4","article-title":"Hepatocellular carcinoma","volume":"400","author":"Vogel","year":"2022","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1016\/S0140-6736(18)30010-2","article-title":"Hepatocellular carcinoma","volume":"391","author":"Forner","year":"2018","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1158\/2159-8290.CD-21-1059","article-title":"Hallmarks of Cancer: New Dimensions","volume":"12","author":"Hanahan","year":"2022","journal-title":"Cancer Discov."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1038\/s42255-022-00659-w","article-title":"Liver cancer metabolism: A hexokinase from the stars","volume":"4","author":"Filliol","year":"2022","journal-title":"Nat. Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.jhep.2013.08.019","article-title":"Genetics of hepatocellular carcinoma: The next generation","volume":"60","author":"Nault","year":"2014","journal-title":"J. Hepatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1038\/s41392-021-00828-5","article-title":"Targeting PI3K\/Akt signal transduction for cancer therapy","volume":"6","author":"He","year":"2021","journal-title":"Signal Transduct. Target Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1007\/s10637-010-9486-3","article-title":"Inhibition of Akt signaling in hepatoma cells induces apoptotic cell death independent of Akt activation status","volume":"29","author":"Buontempo","year":"2011","journal-title":"Investig. New Drugs"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1929","DOI":"10.1002\/cncr.11266","article-title":"Expression and prognostic role of tumor suppressor gene PTEN\/MMAC1\/TEP1 in hepatocellular carcinoma","volume":"97","author":"Hu","year":"2003","journal-title":"Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1002\/cncr.20774","article-title":"Akt phosphorylation is a risk factor for early disease recurrence and poor prognosis in hepatocellular carcinoma","volume":"103","author":"Nakanishi","year":"2005","journal-title":"Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1038\/s41568-019-0216-7","article-title":"The PI3K-AKT network at the interface of oncogenic signalling and cancer metabolism","volume":"20","author":"Hoxhaj","year":"2020","journal-title":"Nat. Rev. Cancer"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1821","DOI":"10.1158\/1078-0432.CCR-21-3208","article-title":"PET-Based Radiogenomics Supports mTOR Pathway Targeting for Hepatocellular Carcinoma","volume":"28","author":"An","year":"2022","journal-title":"Clin. Cancer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2502","DOI":"10.4161\/cc.8.16.9335","article-title":"The Akt kinases: Isoform specificity in metabolism and cancer","volume":"8","author":"Gonzalez","year":"2009","journal-title":"Cell Cycle"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.molmet.2019.08.001","article-title":"The role of skeletal muscle Akt in the regulation of muscle mass and glucose homeostasis","volume":"28","author":"Jaiswal","year":"2019","journal-title":"Mol. Metab."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1186\/1476-4598-11-85","article-title":"Combined targeting of AKT and mTOR synergistically inhibits proliferation of hepatocellular carcinoma cells","volume":"11","author":"Grabinski","year":"2012","journal-title":"Mol. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Grottke, A., Ewald, F., Lange, T., N\u00f6rz, D., Herzberger, C., Bach, J., Grabinski, N., Gr\u00e4ser, L., H\u00f6ppner, F., and Nashan, B. (2016). Downregulation of AKT3 Increases Migration and Metastasis in Triple Negative Breast Cancer Cells by Upregulating S100A4. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0146370"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Hinz, N., Baranowsky, A., Horn, M., Kriegs, M., Sibbertsen, F., Smit, D.J., Clezardin, P., Lange, T., Schinke, T., and J\u00fccker, M. (2021). Knockdown of AKT3 Activates HER2 and DDR Kinases in Bone-Seeking Breast Cancer Cells, Promotes Metastasis In Vivo and Attenuates the TGFbeta\/CTGF Axis. Cells, 10.","DOI":"10.3390\/cells10020430"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7198","DOI":"10.1158\/0008-5472.CAN-14-1490","article-title":"AKT1 and MYC induce distinctive metabolic fingerprints in human prostate cancer","volume":"74","author":"Priolo","year":"2014","journal-title":"Cancer Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.cellsig.2019.03.011","article-title":"AKT2 phosphorylation of hexokinase 2 at T473 promotes tumorigenesis and metastasis in colon cancer cells via NF-kappaB, HIF1alpha, MMP2, and MMP9 upregulation","volume":"58","author":"Li","year":"2019","journal-title":"Cell. Signal."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1093\/abbs\/gmw014","article-title":"Akt3 knockdown induces mitochondrial dysfunction in human cancer cells","volume":"48","author":"Kim","year":"2016","journal-title":"Acta Biochim. Biophys. Sin."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"109976","DOI":"10.1016\/j.celrep.2021.109976","article-title":"Loss of tyrosine catabolic enzyme HPD promotes glutamine anaplerosis through mTOR signaling in liver cancer","volume":"37","author":"Tong","year":"2021","journal-title":"Cell. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1007\/s13402-019-00455-x","article-title":"Apatinib inhibits glycolysis by suppressing the VEGFR2\/AKT1\/SOX5\/GLUT4 signaling pathway in ovarian cancer cells","volume":"42","author":"Chen","year":"2019","journal-title":"Cell. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"118925","DOI":"10.1016\/j.lfs.2020.118925","article-title":"miR-485-3p regulated by MALAT1 inhibits osteosarcoma glycolysis and metastasis by directly suppressing c-MET and AKT3\/mTOR signalling","volume":"268","author":"Wang","year":"2021","journal-title":"Life Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.ccell.2016.07.004","article-title":"Mitochondrial Akt Regulation of Hypoxic Tumor Reprogramming","volume":"30","author":"Chae","year":"2016","journal-title":"Cancer Cell."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1093\/carcin\/bgaa074","article-title":"GLUT5 regulation by AKT1\/3-miR-125b-5p downregulation induces migratory activity and drug resistance in TLR-modified colorectal cancer cells","volume":"41","author":"Park","year":"2020","journal-title":"Carcinogenesis"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1007\/s13105-020-00776-8","article-title":"High expression of NEK2 promotes gastric cancer progression via activating AKT signaling","volume":"77","author":"Wan","year":"2021","journal-title":"J. Physiol. Biochem."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Deng, X., Zhao, J., Qu, L., Duan, Z., Fu, R., Zhu, C., and Fan, D. (2020). Ginsenoside Rh4 suppresses aerobic glycolysis and the expression of PD-L1 via targeting AKT in esophageal cancer. Biochem. Pharmacol., 178.","DOI":"10.1016\/j.bcp.2020.114038"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1016\/j.ccr.2008.11.003","article-title":"Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis","volume":"14","author":"Nogueira","year":"2008","journal-title":"Cancer Cell."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e32213","DOI":"10.7554\/eLife.32213","article-title":"Selective eradication of cancer displaying hyperactive Akt by exploiting the metabolic consequences of Akt activation","volume":"7","author":"Nogueira","year":"2018","journal-title":"Elife"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1038\/s41588-017-0026-3","article-title":"Compartmentalized activities of the pyruvate dehydrogenase complex sustain lipogenesis in prostate cancer","volume":"50","author":"Chen","year":"2018","journal-title":"Nat. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1042\/BST0340217","article-title":"Regulation of the pyruvate dehydrogenase complex","volume":"34","author":"Patel","year":"2006","journal-title":"Biochem. Soc. Trans."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1093\/neuonc\/nou369","article-title":"Ras-mediated modulation of pyruvate dehydrogenase activity regulates mitochondrial reserve capacity and contributes to glioblastoma tumorigenesis","volume":"17","author":"Prabhu","year":"2015","journal-title":"Neuro. Oncol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1928","DOI":"10.1158\/1535-7163.MCT-14-0888","article-title":"The PI3K\/Akt Pathway Regulates Oxygen Metabolism via Pyruvate Dehydrogenase (PDH)-E1alpha Phosphorylation","volume":"14","author":"Cerniglia","year":"2015","journal-title":"Mol. Cancer Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1038\/s42255-020-0172-2","article-title":"We need to talk about the Warburg effect","volume":"2","author":"DeBerardinis","year":"2020","journal-title":"Nat. Metab."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1038\/nrc3038","article-title":"Otto Warburg\u2019s contributions to current concepts of cancer metabolism","volume":"11","author":"Koppenol","year":"2011","journal-title":"Nat. Rev. Cancer"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1038\/nrc1737","article-title":"Mitochondrial tumour suppressors: A genetic and biochemical update","volume":"5","author":"Gottlieb","year":"2005","journal-title":"Nat. Rev. Cancer"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1393","DOI":"10.1111\/j.1742-4658.2007.05686.x","article-title":"Energy metabolism in tumor cells","volume":"274","author":"Saavedra","year":"2007","journal-title":"FEBS J."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.semcancer.2008.11.010","article-title":"Is Akt the \u201cWarburg kinase\u201d?-Akt-energy metabolism interactions and oncogenesis","volume":"19","author":"Robey","year":"2009","journal-title":"Semin. Cancer Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4896","DOI":"10.1158\/0008-5472.CAN-21-2647","article-title":"Cancer Signaling Drives Cancer Metabolism: AKT and the Warburg Effect","volume":"81","author":"Hosios","year":"2021","journal-title":"Cancer Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1038\/s41571-023-00812-8","article-title":"Novel ICI-TKI combination improves HCC outcomes","volume":"20","author":"Killock","year":"2023","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_41","first-page":"1","article-title":"Sorafenib resistance in hepatocarcinoma: Role of hypoxia-inducible factors","volume":"50","author":"Fondevila","year":"2018","journal-title":"Exp. Mol. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1331","DOI":"10.1007\/s00262-018-2195-z","article-title":"Targeting tumor-associated acidity in cancer immunotherapy","volume":"67","author":"Lacroix","year":"2018","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Wang, J.X., Choi, S.Y., Niu, X., Kang, N., Xue, H., Killam, J., and Wang, Y. (2020). Lactic Acid and an Acidic Tumor Microenvironment suppress Anticancer Immunity. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21218363"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e1445452","DOI":"10.1080\/2162402X.2018.1445452","article-title":"pH regulators to target the tumor immune microenvironment in human hepatocellular carcinoma","volume":"7","author":"Kuchuk","year":"2018","journal-title":"Oncoimmunology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2058","DOI":"10.1056\/NEJMoa2214131","article-title":"Capivasertib in Hormone Receptor-Positive Advanced Breast Cancer","volume":"388","author":"Turner","year":"2023","journal-title":"N. Engl. J. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/4\/2168\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:58:31Z","timestamp":1760104711000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/4\/2168"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,11]]},"references-count":45,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2024,2]]}},"alternative-id":["ijms25042168"],"URL":"https:\/\/doi.org\/10.3390\/ijms25042168","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,11]]}}}