{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T13:39:23Z","timestamp":1765805963318,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2020,4,27]],"date-time":"2020-04-27T00:00:00Z","timestamp":1587945600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["POCI-01-0145-FEDER-030388 - PTDC\/SAU-SER\/30388\/2017"],"award-info":[{"award-number":["POCI-01-0145-FEDER-030388 - PTDC\/SAU-SER\/30388\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>The identification of noninvasive biomarkers able to detect renal cell carcinoma (RCC) at an early stage remains an unmet clinical need. The recognition that altered metabolism is a core hallmark of cancer boosted metabolomic studies focused in the search for cancer biomarkers. The present work aims to evaluate the performance of the volatile metabolites present in the extracellular medium to discriminate RCC cell lines with distinct histological subtypes (clear cell and papillary) and metastatic potential from non-tumorigenic renal cells. Hence, volatile organic compounds (VOCs) and volatile carbonyl compounds (VCCs) were extracted by headspace solid-phase microextraction (HS-SPME) and analyzed by gas chromatography\u2013mass spectrometry (GC\u2013MS). Multivariate and univariate analysis unveiled a panel of metabolites responsible for the separation between groups, mostly belonging to ketones, alcohols, alkanes and aldehydes classes. Some metabolites were found similarly altered for all RCC cell lines compared to non-tumorigenic cells, namely 2-ethylhexanol, tetradecane, formaldehyde, acetone (increased) and cyclohexanone and acetaldehyde (decreased). Furthermore, significantly altered levels of cyclohexanol, decanal, decane, dodecane and 4-methylbenzaldehyde were observed in all metastatic RCC cell lines when compared with the non-metastatic ones. Moreover, some alterations in the volatile composition were also observed between RCC histological subtypes. Overall, our results demonstrate the potential of volatile profiling for identification of noninvasive candidate biomarkers for early RCC diagnosis.<\/jats:p>","DOI":"10.3390\/metabo10050174","type":"journal-article","created":{"date-parts":[[2020,4,29]],"date-time":"2020-04-29T01:29:15Z","timestamp":1588123755000},"page":"174","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Volatilomics Reveals Potential Biomarkers for Identification of Renal Cell Carcinoma: An In Vitro Approach"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5050-2864","authenticated-orcid":false,"given":"Filipa","family":"Amaro","sequence":"first","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9719-8364","authenticated-orcid":false,"given":"Joana","family":"Pinto","sequence":"additional","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"}]},{"given":"S\u00edlvia","family":"Rocha","sequence":"additional","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"},{"name":"Master in Oncology, Institute of Biomedical Sciences Abel Salazar\u2013University of Porto (ICBAS-UP), 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4073-2121","authenticated-orcid":false,"given":"Ana Margarida","family":"Ara\u00fajo","sequence":"additional","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4231-5532","authenticated-orcid":false,"given":"Vera","family":"Miranda-Gon\u00e7alves","sequence":"additional","affiliation":[{"name":"Cancer Biology &amp; Epigenetics Group, Research Centre (CI-IPOP) Portuguese Oncology Institute of Porto (IPO Porto), 4200-072 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4186-5345","authenticated-orcid":false,"given":"Carmen","family":"Jer\u00f3nimo","sequence":"additional","affiliation":[{"name":"Cancer Biology &amp; Epigenetics Group, Research Centre (CI-IPOP) Portuguese Oncology Institute of Porto (IPO Porto), 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology-Biomedical Sciences Institute (ICBAS), University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3171-4666","authenticated-orcid":false,"given":"Rui","family":"Henrique","sequence":"additional","affiliation":[{"name":"Cancer Biology &amp; Epigenetics Group, Research Centre (CI-IPOP) Portuguese Oncology Institute of Porto (IPO Porto), 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology-Biomedical Sciences Institute (ICBAS), University of Porto, 4050-313 Porto, Portugal"},{"name":"Department of Pathology, Portuguese Oncology Institute of Porto (IPO Porto), 4200-072 Porto, Portugal"}]},{"given":"Maria de Lourdes","family":"Bastos","sequence":"additional","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9884-4751","authenticated-orcid":false,"given":"M\u00e1rcia","family":"Carvalho","sequence":"additional","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"},{"name":"UFP Energy, Environment and Health Research Unit (FP-ENAS), University Fernando Pessoa, 349, 4249-004 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7395-5700","authenticated-orcid":false,"given":"Paula","family":"Guedes de Pinho","sequence":"additional","affiliation":[{"name":"UCIBIO, REQUIMTE, Laboratory of Toxicology, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1016\/j.eururo.2019.02.011","article-title":"European Association of Urology Guidelines on Renal Cell Carcinoma: The 2019 Update","volume":"75","author":"Ljungberg","year":"2019","journal-title":"Eur. Urol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"663","DOI":"10.3389\/fonc.2019.00663","article-title":"Urine Metabolomics for Renal Cell Carcinoma (RCC) Prediction: Tryptophan Metabolism as an Important Pathway in RCC","volume":"9","author":"Liu","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1590\/0100-3984.2013.1927","article-title":"Renal cell carcinoma: histological classification and correlation with imaging findings*","volume":"48","author":"Muglia","year":"2015","journal-title":"Radiol. Bras."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1097\/00000478-200305000-00005","article-title":"Comparisons of outcome and prognostic features among histologic subtypes of renal cell carcinoma","volume":"27","author":"Cheville","year":"2003","journal-title":"Am. J. Surg. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1210","DOI":"10.1007\/s11306-014-0659-5","article-title":"Biomarkers in renal cell carcinoma: A metabolomics approach","volume":"10","author":"Monteiro","year":"2014","journal-title":"Metabolomics"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e16309","DOI":"10.1097\/MD.0000000000016309","article-title":"A comparison of the prognosis of papillary and clear cell renal cell carcinoma: Evidence from a meta-analysis","volume":"98","author":"Deng","year":"2019","journal-title":"Medicine"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"59189","DOI":"10.18632\/oncotarget.10830","article-title":"Prediction and diagnosis of renal cell carcinoma using nuclear magnetic resonance-based serum metabolomics and self-organizing maps","volume":"7","author":"Zheng","year":"2016","journal-title":"Oncotarget"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"744","DOI":"10.1016\/j.bbadis.2016.12.011","article-title":"Integrated transcriptomic and metabolomic analysis shows that disturbances in metabolism of tumor cells contribute to poor survival of RCC patients","volume":"1863","author":"Tohge","year":"2017","journal-title":"Biochim. et Biophys. Acta (BBA) - Mol. Basis Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"620","DOI":"10.21037\/tcr.2017.05.19","article-title":"Renal cell carcinoma: the search for a reliable biomarker","volume":"6","author":"Farber","year":"2017","journal-title":"Transl. Cancer Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1080\/14737159.2020.1704259","article-title":"Role of metabolomics-derived biomarkers to identify renal cell carcinoma: a comprehensive perspective of the past ten years and advancements","volume":"20","author":"Gupta","year":"2019","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1038\/sj.ki.5000065","article-title":"Kidney cancer: Identification of novel targets for therapy","volume":"69","author":"Weiss","year":"2006","journal-title":"Kidney Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1080\/08998280.2005.11928093","article-title":"Renal Cell Carcinoma: Diagnosis and Treatment, 1994\u20132003","volume":"18","author":"Hutson","year":"2005","journal-title":"Bayl. Univ. Med Cent. Proc."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1007\/978-3-319-47656-8_9","article-title":"Applications of Metabolomics in Cancer Studies","volume":"965","author":"Armitage","year":"2017","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"167","DOI":"10.3390\/diseases3030167","article-title":"Metabolomic Approaches in Cancer Epidemiology","volume":"3","author":"Verma","year":"2015","journal-title":"Diseases"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1038\/nrneph.2017.59","article-title":"Metabolic reprogramming in clear cell renal cell carcinoma","volume":"13","author":"Wettersten","year":"2017","journal-title":"Nat. Rev. Nephrol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.trsl.2016.07.018","article-title":"Renal cell carcinoma: a critical analysis of metabolomic biomarkers emerging from current model systems","volume":"180","author":"Rodrigues","year":"2017","journal-title":"Transl. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3859","DOI":"10.1007\/s00216-018-1059-x","article-title":"Metabolomic study of human tissue and urine in clear cell renal carcinoma by LC-HRMS and PLS-DA","volume":"410","author":"Bonifay","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1007\/s00432-011-1134-6","article-title":"Application of ex vivo 1H NMR metabonomics to the characterization and possible detection of renal cell carcinoma metastases","volume":"138","author":"Gao","year":"2012","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Falegan, O.S., Ball, M., Shaykhutdinov, R.A., Pieroraio, P.M., Farshidfar, F., Vogel, H.J., Allaf, M.E., and Hyndman, M.E. (2017). Urine and Serum Metabolomics Analyses May Distinguish between Stages of Renal Cell Carcinoma. Metabolites, 7.","DOI":"10.3390\/metabo7010006"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"981458","DOI":"10.1155\/2015\/981458","article-title":"Current Challenges in Volatile Organic Compounds Analysis as Potential Biomarkers of Cancer","volume":"2015","author":"Schmidt","year":"2015","journal-title":"J. Biomark."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5539","DOI":"10.1038\/s41598-018-23847-9","article-title":"Discrimination between the human prostate normal and cancer cell exometabolome by GC-MS","volume":"8","author":"Lima","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Jia, Z., Patra, A., Kutty, V.K., and Venkatesan, V. (2019). Critical Review of Volatile Organic Compound Analysis in Breath and In Vitro Cell Culture for Detection of Lung Cancer. Metabolites, 9.","DOI":"10.3390\/metabo9030052"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"5131","DOI":"10.1021\/acsomega.7b02035","article-title":"Detection of Lung Cancer: Concomitant Volatile Organic Compounds and Metabolomic Profiling of Six Cancer Cell Lines of Different Histological Origins","volume":"3","author":"Jia","year":"2018","journal-title":"ACS Omega"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1007\/s11306-018-1361-9","article-title":"Volatile metabolomic signature of bladder cancer cell lines based on gas chromatography\u2013mass spectrometry","volume":"14","author":"Rodrigues","year":"2018","journal-title":"Metabolomics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"43969","DOI":"10.1038\/srep43969","article-title":"Volatile metabolomic signature of human breast cancer cell lines","volume":"7","author":"Silva","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2092","DOI":"10.1111\/jcmm.13132","article-title":"GC-MS metabolomics-based approach for the identification of a potential VOC-biomarker panel in the urine of renal cell carcinoma patients","volume":"21","author":"Monteiro","year":"2017","journal-title":"J. Cell. Mol. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"68","DOI":"10.3892\/br.2016.686","article-title":"Urinary volatile organic compounds as potential biomarkers for renal cell carcinoma","volume":"5","author":"Wang","year":"2016","journal-title":"Biomed. Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1038\/s41416-019-0585-4","article-title":"Identification of a biomarker panel for improvement of prostate cancer diagnosis by volatile metabolic profiling of urine","volume":"121","author":"Lima","year":"2019","journal-title":"Br. J. Cancer"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1016\/j.ijnurstu.2012.01.015","article-title":"Effect size estimation: Methods and examples","volume":"49","author":"Berben","year":"2012","journal-title":"Int. J. Nurs. Stud."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"bpz014","DOI":"10.1093\/biomethods\/bpz014","article-title":"Digging deeper into volatile organic compounds associated with cancer","volume":"4","author":"Janfaza","year":"2019","journal-title":"Boil. Methods Protoc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1172","DOI":"10.1016\/j.juro.2011.11.105","article-title":"Status of Oxidative Stress in Patients With Renal Cell Carcinoma","volume":"187","author":"Ganesamoni","year":"2012","journal-title":"J. Urol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1089\/ars.2015.6569","article-title":"Renal Carcinogenesis, Tumor Heterogeneity, and Reactive Oxygen Species: Tactics Evolved","volume":"25","author":"Shanmugasundaram","year":"2016","journal-title":"Antioxid. Redox Signal."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/j.intimp.2010.12.023","article-title":"Superoxide dismutase and cytochrome P450 isoenzymes might be associated with higher risk of renal cell carcinoma in male patients","volume":"11","author":"Kraus","year":"2011","journal-title":"Int. Immunopharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1016\/j.freeradbiomed.2005.09.026","article-title":"Lipid peroxidation and renal cell carcinoma: further supportive evidence and new mechanistic insights","volume":"40","author":"Castelao","year":"2006","journal-title":"Free. Radic. Boil. Med."},{"key":"ref_35","first-page":"2316","article-title":"Identification of metabolism-associated genes and pathways involved in different stages of clear cell renal cell carcinoma","volume":"15","author":"Li","year":"2017","journal-title":"Oncol. Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"966","DOI":"10.1038\/sj.bjc.6602053","article-title":"Cytochrome P450 CYP1B1 activity in renal cell carcinoma","volume":"91","author":"McFadyen","year":"2004","journal-title":"Br. J. Cancer"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1271\/bbb.110760","article-title":"Urinary Volatile Compounds as Biomarkers for Lung Cancer","volume":"76","author":"Hanai","year":"2012","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_38","first-page":"243.e21","article-title":"Furan and p-xylene as candidate biomarkers for prostate cancer","volume":"36","author":"Iribar","year":"2018","journal-title":"Urol. Oncol. Semin. Orig. Investig."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/j.yexmp.2015.03.012","article-title":"The activity of class I, II, III and IV alcohol dehydrogenase isoenzymes and aldehyde dehydrogenase in renal cell carcinoma","volume":"98","author":"Orywal","year":"2015","journal-title":"Exp. Mol. Pathol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1007\/s10238-016-0408-3","article-title":"Alcohol dehydrogenase and aldehyde dehydrogenase in malignant neoplasms","volume":"17","author":"Orywal","year":"2016","journal-title":"Clin. Exp. Med."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.semnephrol.2018.01.006","article-title":"Metabolomics and Metabolic Reprogramming in Kidney Cancer","volume":"38","author":"Weiss","year":"2018","journal-title":"Semin. Nephrol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1016\/j.euf.2018.01.016","article-title":"Metabolic and Lipidomic Reprogramming in Renal Cell Carcinoma Subtypes Reflects Regions of Tumor Origin","volume":"5","author":"Schaeffeler","year":"2019","journal-title":"Eur. Urol. Focus"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"15635","DOI":"10.1038\/s41598-019-52059-y","article-title":"LC-MS based metabolomic profiling for renal cell carcinoma histologic subtypes","volume":"9","author":"Jing","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.ccell.2015.12.004","article-title":"An Integrated Metabolic Atlas of Clear Cell Renal Cell Carcinoma","volume":"29","author":"Hakimi","year":"2016","journal-title":"Cancer Cell"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1016\/j.cell.2011.02.013","article-title":"Hallmarks of Cancer: The Next Generation","volume":"144","author":"Hanahan","year":"2011","journal-title":"Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/s12943-016-0565-8","article-title":"Choosing the right cell line for renal cell cancer research","volume":"15","author":"Brodaczewska","year":"2016","journal-title":"Mol. Cancer"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1038\/ki.1994.6","article-title":"HK-2: An immortalized proximal tubule epithelial cell line from normal adult human kidney","volume":"45","author":"Ryan","year":"1994","journal-title":"Kidney Int."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.toxlet.2018.05.028","article-title":"Analysis of extracellular metabolome by HS-SPME\/GC\u2013MS: Optimization and application in a pilot study to evaluate galactosamine-induced hepatotoxicity","volume":"295","author":"Araujo","year":"2018","journal-title":"Toxicol. Lett."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Pluskal, T., Castillo, S., Villar-Briones, A., and Ore\u0161i\u010d, M. (2010). MZmine 2: Modular framework for processing, visualizing, and analyzing mass spectrometry-based molecular profile data. BMC Bioinform., 11.","DOI":"10.1186\/1471-2105-11-395"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"W486","DOI":"10.1093\/nar\/gky310","article-title":"MetaboAnalyst 4.0: Towards more transparent and integrative metabolomics analysis","volume":"46","author":"Chong","year":"2018","journal-title":"Nucleic Acids Res."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/10\/5\/174\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:51:28Z","timestamp":1760363488000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/10\/5\/174"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,27]]},"references-count":50,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["metabo10050174"],"URL":"https:\/\/doi.org\/10.3390\/metabo10050174","relation":{},"ISSN":["2218-1989"],"issn-type":[{"type":"electronic","value":"2218-1989"}],"subject":[],"published":{"date-parts":[[2020,4,27]]}}}