{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T19:01:28Z","timestamp":1771268488920,"version":"3.50.1"},"reference-count":58,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,3,19]],"date-time":"2022-03-19T00:00:00Z","timestamp":1647648000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Lung cancer (LC) is the leading cause of cancer-related death worldwide. Although the diagnosis and treatment of non-small cell lung cancer (NSCLC), which accounts for approximately 80% of LC cases, have greatly improved in the past decade, there is still an urgent need to find more sensitive and specific screening methods. Recently, new molecular biomarkers are emerging as potential non-invasive diagnostic agents to screen NSCLC, including multiple microRNAs (miRNAs) that show an unusual expression profile. Moreover, peripheral blood mononuclear cells\u2019 (PBMCs) miRNA profile could be linked with NSCLC and used for diagnosis. We developed a molecular beacon (MB)-based miRNA detection strategy for NSCLC. Following PBMCs isolation and screening of the expression profile of a panel of miRNA by RT-qPCR, we designed a MB targeting of up-regulated miR-21-5p. This MB 21-5p was characterized by FRET-melting, CD, NMR and native PAGE, allowing the optimization of an in-situ approach involving miR-21-5p detection in PBMCs via MB. Data show the developed MB approach potential for miR-21-5p detection in PBMCs from clinical samples towards NSCLC.<\/jats:p>","DOI":"10.3390\/ijms23063330","type":"journal-article","created":{"date-parts":[[2022,3,20]],"date-time":"2022-03-20T21:30:14Z","timestamp":1647811814000},"page":"3330","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Molecular Beacon for Detection miRNA-21 as a Biomarker of Lung Cancer"],"prefix":"10.3390","volume":"23","author":[{"given":"Daniela","family":"Alexandre","sequence":"first","affiliation":[{"name":"CICS-UBI\u2014Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Universidade da Beira Interior, Av. Infante D. 5 Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"given":"Bernardo","family":"Teixeira","sequence":"additional","affiliation":[{"name":"CICS-UBI\u2014Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Universidade da Beira Interior, Av. Infante D. 5 Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4196-006X","authenticated-orcid":false,"given":"Andr\u00e9","family":"Rico","sequence":"additional","affiliation":[{"name":"CICS-UBI\u2014Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Universidade da Beira Interior, Av. Infante D. 5 Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9281-2003","authenticated-orcid":false,"given":"Salete","family":"Valente","sequence":"additional","affiliation":[{"name":"Servi\u00e7o de Pneumologia do Centro Hospitalar Universit\u00e1rio Cova da Beira (CHUCB), 6200-506 Covilh\u00e3, Portugal"}]},{"given":"Ana","family":"Craveiro","sequence":"additional","affiliation":[{"name":"Servi\u00e7o de Pneumologia do Centro Hospitalar Universit\u00e1rio Cova da Beira (CHUCB), 6200-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5255-7095","authenticated-orcid":false,"given":"Pedro V.","family":"Baptista","sequence":"additional","affiliation":[{"name":"UCIBIO, Department of Life Sciences, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"},{"name":"i4HB, Associate Laboratory\u2013Institute for Health and Bioeconomy, FCT-NOVA, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6630-1242","authenticated-orcid":false,"given":"Carla","family":"Cruz","sequence":"additional","affiliation":[{"name":"CICS-UBI\u2014Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Universidade da Beira Interior, Av. Infante D. 5 Henrique, 6200-506 Covilh\u00e3, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6861","DOI":"10.1158\/0008-5472.CAN-08-2827","article-title":"The History of Cancer Research: Introducing an AACR Centennial Series","volume":"68","author":"Weinstein","year":"2008","journal-title":"Cancer Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ccm.2019.10.001","article-title":"Lung Cancer 2020: Epidemiology, Etiology, and Prevention","volume":"41","author":"Bade","year":"2020","journal-title":"Clin. Chest Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"8","DOI":"10.5334\/aogh.2419","article-title":"Global Epidemiology of Lung Cancer","volume":"85","author":"Barta","year":"2019","journal-title":"Ann. Glob. Health"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1146\/annurev.med.54.101601.152202","article-title":"Molecular Genetics of Lung Cancer","volume":"54","author":"Sekido","year":"2003","journal-title":"Annu. Rev. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1767","DOI":"10.1016\/j.rmed.2010.08.005","article-title":"Staging of non-small cell lung cancer (NSCLC): A review","volume":"104","author":"Tsim","year":"2010","journal-title":"Respir. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.semcancer.2017.09.005","article-title":"Lung cancer epigenetics: From knowledge to applications","volume":"51","author":"Duruisseaux","year":"2018","journal-title":"Semin. Cancer Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.soc.2016.02.003","article-title":"Classification and Pathology of Lung Cancer","volume":"25","author":"Zheng","year":"2016","journal-title":"Surg. Oncol. Clin. N. Am."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1623","DOI":"10.1016\/j.mayocp.2019.01.013","article-title":"Non-Small Cell Lung Cancer: Epidemiology, Screening, Diagnosis, and Treatment","volume":"94","author":"Duma","year":"2019","journal-title":"Mayo Clin. Proc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1136\/thoraxjnl-2011-200714","article-title":"Early diagnosis of lung cancer: Evaluation of a community-based social marketing intervention","volume":"67","author":"Athey","year":"2011","journal-title":"Thorax"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"170070","DOI":"10.1098\/rsob.170070","article-title":"Progress and prospects of early detection in lung cancer","volume":"7","author":"Knight","year":"2017","journal-title":"Open Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1248","DOI":"10.1016\/j.jtho.2018.05.030","article-title":"Liquid Biopsy for Advanced Non-Small Cell Lung Cancer (NSCLC): A Statement Paper from the IASLC","volume":"13","author":"Rolfo","year":"2018","journal-title":"J. Thorac. Oncol."},{"key":"ref_13","first-page":"S540","article-title":"Exhaled breath analysis for lung cancer","volume":"5","author":"Dent","year":"2013","journal-title":"J. Thorac. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.hoc.2016.08.006","article-title":"Lung Cancer Biomarkers","volume":"31","author":"Villalobos","year":"2017","journal-title":"Hematol. Clin. N. Am."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1111\/1759-7714.13337","article-title":"MicroRNA-based biomarkers for diagnosis of non-small cell lung cancer (NSCLC)","volume":"11","author":"Liao","year":"2020","journal-title":"Thorac. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3119","DOI":"10.21037\/jtd.2018.05.32","article-title":"miRNAs as biomarkers and for the early detection of non-small cell lung cancer (NSCLC)","volume":"10","author":"Han","year":"2018","journal-title":"J. Thorac. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"R17","DOI":"10.1093\/hmg\/ddl046","article-title":"Non-coding RNA","volume":"15","author":"Mattick","year":"2006","journal-title":"Hum. Mol. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"De Sousa, M.C., Gjorgjieva, M., Dolicka, D., Sobolewski, C., and Foti, M. (2019). Deciphering miRNAs\u2019 Action through miRNA Editing. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20246249"},{"key":"ref_19","first-page":"175","article-title":"Extracellular\/Circulating MicroRNAs: Release Mechanisms, Functions and Challenges","volume":"10","author":"Sohel","year":"2016","journal-title":"Achieve Life Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"5451","DOI":"10.1002\/jcp.27486","article-title":"An overview of microRNAs: Biology, functions, therapeutics, and analysis methods","volume":"234","author":"Saliminejad","year":"2019","journal-title":"J. Cell. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Wu, K.-L., Tsai, Y.-M., Lien, C.-T., Kuo, P.-L., and Hung, A.J.-Y. (2019). The Roles of MicroRNA in Lung Cancer. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20071611"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"643942","DOI":"10.3389\/fcell.2021.643942","article-title":"Frontiers of MicroRNA Signature in Non-small Cell Lung Cancer","volume":"9","author":"Zhu","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"193","DOI":"10.3389\/fgene.2016.00193","article-title":"Circulating Plasma MicroRNAs As Diagnostic Markers for NSCLC","volume":"7","author":"Hou","year":"2016","journal-title":"Front. Genet."},{"key":"ref_24","first-page":"321","article-title":"Serum MiRNA as Predictive and Prognosis Biomarker in Advanced Stage Non-small Cell Lung Cancer in Indonesia","volume":"23","author":"Hanafi","year":"2020","journal-title":"Zhongguo Fei Ai Za Zhi = Chin. J. Lung Cancer"},{"key":"ref_25","first-page":"19","article-title":"Altered expression of miRNA-155 and miRNA-21 and higher numbers of T cells in the peripheral blood of early diagnosed lung cancer patients","volume":"4","author":"Salem","year":"2020","journal-title":"Int. J. Cancer Biomed. Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Mosallaei, M., Ehtesham, N., Rahimirad, S., Saghi, M., Vatandoost, N., and Khosravi, S. (2020). PBMCs: A new source of diagnostic and prognostic biomarkers. Arch. Physiol. Biochem., 1\u20137.","DOI":"10.1080\/13813455.2020.1752257"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"367157","DOI":"10.1155\/2014\/367157","article-title":"MicroRNA Expression Profiling in PBMCs: A Potential Diagnostic Biomarker of Chronic Hepatitis C","volume":"2014","author":"Chang","year":"2014","journal-title":"Dis. Markers"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wang, C., Yang, S., Sun, G., Tang, X., Lu, S., Neyrolles, O., and Gao, Q. (2011). Comparative miRNA Expression Profiles in Individuals with Latent and Active Tuberculosis. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0025832"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1080\/15592294.2019.1676613","article-title":"Identification of PBMC-expressed miRNAs for rheumatoid arthritis","volume":"15","author":"Zhu","year":"2020","journal-title":"Epigenetics"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1038\/labinvest.2015.88","article-title":"Differential miRNA expressions in peripheral blood mononuclear cells for diagnosis of lung cancer","volume":"95","author":"Ma","year":"2015","journal-title":"Lab. Investig."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1016\/j.cbpa.2004.08.010","article-title":"Molecular beacons","volume":"8","author":"Tan","year":"2004","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1002\/anie.200800370","article-title":"Molecular Engineering of DNA: Molecular Beacons","volume":"48","author":"Wang","year":"2009","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"122037","DOI":"10.1016\/j.talanta.2020.122037","article-title":"Aptamer-based approaches to detect nucleolin in prostate cancer","volume":"226","author":"Miranda","year":"2021","journal-title":"Talanta"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Carvalho, J., Lopes-Nunes, J., Figueiredo, J., Santos, T., Miranda, A., Riscado, M., Sousa, F., Duarte, A.P., Socorro, S., and Tomaz, C.T. (2021). Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection. Sensors, 21.","DOI":"10.22541\/au.161652048.85831942\/v1"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.biomaterials.2015.03.014","article-title":"In situ single step detection of exosome microRNA using molecular beacon","volume":"54","author":"Lee","year":"2015","journal-title":"Biomaterials"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.mcp.2012.07.001","article-title":"Novel molecular beacons to monitor microRNAs in non-small-cell lung cancer","volume":"26","author":"Yao","year":"2012","journal-title":"Mol. Cell. Probes"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3625823","DOI":"10.1155\/2018\/3625823","article-title":"Quantitative Detection of miRNA-21 Expression in Tumor Cells and Tissues Based on Molecular Beacon","volume":"2018","author":"Liu","year":"2018","journal-title":"Int. J. Anal. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"105277","DOI":"10.1016\/j.microc.2020.105277","article-title":"A fluorescence-readout method for miRNA-155 detection with double-hairpin molecular beacon based on quadruplex DNA structure","volume":"158","author":"Shahsavar","year":"2020","journal-title":"Microchem. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3539","DOI":"10.1007\/s00018-018-2877-x","article-title":"Overview upon miR-21 in lung cancer: Focus on NSCLC","volume":"75","author":"Magdo","year":"2018","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1016\/j.cca.2010.02.074","article-title":"MicroRNA-21 (miR-21) represses tumor suppressor PTEN and promotes growth and invasion in non-small cell lung cancer (NSCLC)","volume":"411","author":"Zhang","year":"2010","journal-title":"Clin. Chim. Acta"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/s11010-012-1443-3","article-title":"MicroRNA-21 (miR-21) expression promotes growth, metastasis, and chemo- or radioresistance in non-small cell lung cancer cells by targeting PTEN","volume":"372","author":"Liu","year":"2012","journal-title":"Mol. Cell. Biochem."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Shen, H., Zhu, F., Liu, J., Xu, T., Pei, D., Wang, R., Qian, Y., Li, Q., Wang, L., and Shi, Z. (2014). Alteration in Mir-21\/PTEN Expression Modulates Gefitinib Resistance in Non-Small Cell Lung Cancer. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0103305"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1159\/000488831","article-title":"Effect of miR-21 on Apoptosis in Lung Cancer Cell Through Inhibiting the PI3K\/ Akt\/NF-\u03baB Signaling Pathway in Vitro and in Vivo","volume":"46","author":"Zhou","year":"2018","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_44","first-page":"e85456","article-title":"Ectopic Expression of miRNA-21 and miRNA-205 in Non-Small Cell Lung Cancer","volume":"12","author":"Mohamadnia","year":"2019","journal-title":"Int. J. Cancer Manag."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1016\/j.bbrc.2013.09.111","article-title":"MiR-92b regulates the cell growth, cisplatin chemosensitivity of A549 non small cell lung cancer cell line and target PTEN","volume":"440","author":"Li","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"492","DOI":"10.3892\/or.2013.2453","article-title":"miR-150, p53 protein and relevant miRNAs consist of a regulatory network in NSCLC tumorigenesis","volume":"30","author":"Wang","year":"2013","journal-title":"Oncol. Rep."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"84508","DOI":"10.18632\/oncotarget.13022","article-title":"MiR-21 and MiR-155 promote non-small cell lung cancer progression by downregulating SOCS1, SOCS6, and PTEN","volume":"7","author":"Xue","year":"2016","journal-title":"Oncotarget"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1002\/1878-0261.12420","article-title":"miR-181a\/b therapy in lung cancer: Reality or myth?","volume":"13","author":"Braicu","year":"2019","journal-title":"Mol. Oncol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/j.trsl.2015.05.009","article-title":"Plasma circulating microRNA-944 and microRNA-3662 as potential histologic type-specific early lung cancer biomarkers","volume":"166","author":"Krawczyk","year":"2015","journal-title":"Transl. Res."},{"key":"ref_50","first-page":"4655","article-title":"miR-21-5p induces cell proliferation by targeting TGFBI in non-small cell lung cancer cells","volume":"16","author":"Yan","year":"2018","journal-title":"Exp. Ther. Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"4018","DOI":"10.1038\/s41598-020-60157-5","article-title":"Evaluation of Rationally Designed Label-free Stem-loop DNA Probe Opening in the Presence of miR-21 by Circular Dichroism and Fluorescence Techniques","volume":"10","author":"Farahani","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1093\/nar\/gkp026","article-title":"Circular dichroism and conformational polymorphism of DNA","volume":"37","author":"Kypr","year":"2009","journal-title":"Nucleic Acids Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"4840","DOI":"10.1021\/bi00365a018","article-title":"NMR study of a synthetic DNA hairpin","volume":"25","author":"Ikuta","year":"1986","journal-title":"Biochemistry"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"3938","DOI":"10.1093\/nar\/27.19.3938","article-title":"Structural characterisation of a uracil containing hairpin DNA by NMR and molecular dynamics","volume":"27","author":"Ghosh","year":"1999","journal-title":"Nucleic Acids Res."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1093\/nar\/21.3.537","article-title":"Structure of a small RNA hairpin","volume":"21","author":"Davis","year":"1993","journal-title":"Nucleic Acids Res."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Chirayil, S., Wu, Q., Amezcua, C., and Luebke, K.J. (2014). NMR Characterization of an Oligonucleotide Model of the MiR-21 Pre-Element. PLoS ONE, 9.","DOI":"10.2210\/pdb2mnc\/pdb"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"5485","DOI":"10.1093\/nar\/21.23.5485","article-title":"NMR evidence for the RNA stem-loop structure involved in the transcription attenuation of E.coli trp operon","volume":"21","author":"Ramesh","year":"1993","journal-title":"Nucleic Acids Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/1748-7188-6-26","article-title":"ViennaRNA Package 2.0","volume":"6","author":"Lorenz","year":"2011","journal-title":"Algorithms Mol. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3330\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:39:37Z","timestamp":1760135977000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3330"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,19]]},"references-count":58,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["ijms23063330"],"URL":"https:\/\/doi.org\/10.3390\/ijms23063330","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,19]]}}}