{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T03:58:04Z","timestamp":1787025484055,"version":"build-2736575974"},"reference-count":50,"publisher":"IMR Press","issue":"4","license":[{"start":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T00:00:00Z","timestamp":1776988800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Health and Medical Research Council of Australia","award":["APP2011845"],"award-info":[{"award-number":["APP2011845"]}]},{"name":"Australian Government Research Training Program (RTP) Scholarship"},{"name":"Supplementary scholarships from The Hospital Research Foundation Group"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Front. Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background: Cellular stress can arise from external and internal stimuli and induce mechanisms that either maintain survival or activate cell death pathways. Nuclear factor kappa B (NF-\u03baB) is a key transcription factor that mediates a range of responses to cellular stress, including senescence, DNA repair, and inflammation in breast cancer; however, there are currently limited biomarkers that identify its activation. CalRexinTM, a monoclonal antibody that targets the human calcitonin receptor, was investigated as a novel biomarker for cell stress associated with NF-\u03baB activation. Methods: Triple-negative breast cancer cell lines MDA-MB-231 and MDA-MB-468 were treated with lipopolysaccharide (LPS) to induce NF-\u03baB-mediated cellular stress or treated with the cytotoxin staurosporine to induce apoptosis as a positive control. CalRexinTM and Annexin V positivity were investigated by flow cytometry and immunofluorescence. Results: Flow cytometry analysis revealed a dose- and time-dependent increase in the mean fluorescence intensity of CalRexinTM following LPS treatment of breast cancer cells. The mean fluorescence intensity of Annexin V, as a marker of apoptosis was not increased by LPS treatment and the cells remained viable. Immunofluorescence imaging demonstrated CalRexinTM positivity in LPS-stimulated cells accompanied by nuclear translocation of NF-\u03baB. Conclusion: Increased expression of calcitonin receptor, identified using the novel CalRexinTM antibody, has potential use as a biomarker for NF-\u03baB-mediated cellular stress in breast cancer cells.<\/jats:p>","DOI":"10.31083\/fbl50148","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T09:29:27Z","timestamp":1777454967000},"source":"Crossref","is-referenced-by-count":1,"title":["Calcitonin Receptor is a Novel Biomarker for NF-\u03baB-Mediated Cellular Stress in Triple-Negative Breast Cancer Cells"],"prefix":"10.31083","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9103-2926","authenticated-orcid":true,"given":"Hanieh","family":"Heydarlou","sequence":"first","affiliation":[{"name":"School of Medicine, College of Health, Adelaide University, The Queen Elizabeth Hospital, Woodville South, SA 5011, Australia"},{"name":"Robinson Research Institute, Adelaide University, Adelaide, SA 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6840-2653","authenticated-orcid":true,"given":"Reza","family":"Maleki","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Health, Adelaide University, The Queen Elizabeth Hospital, Woodville South, SA 5011, Australia"},{"name":"Robinson Research Institute, Adelaide University, Adelaide, SA 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3937-1621","authenticated-orcid":true,"given":"Peter J","family":"Wookey","sequence":"additional","affiliation":[{"name":"Department of Medicine-Austin, University of Melbourne, Austin Health, Heidelberg, VIC 3084, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2958-3492","authenticated-orcid":true,"given":"Eric","family":"Smith","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Health, Adelaide University, The Queen Elizabeth Hospital, Woodville South, SA 5011, Australia"},{"name":"Medical Oncology, Basil Hetzel Institute, The Queen Elizabeth Hospital, Woodville South, SA 5011, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3116-2902","authenticated-orcid":true,"given":"Wendy V","family":"Ingman","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Health, Adelaide University, The Queen Elizabeth Hospital, Woodville South, SA 5011, Australia"},{"name":"Robinson Research Institute, Adelaide University, Adelaide, SA 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,4,24]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Twayana KS, Ravanan P. Eukaryotic cell survival mechanisms: Disease relevance and therapeutic intervention. Life Sciences. 2018; 205: 73\u201390.","DOI":"10.1016\/j.lfs.2018.05.002"},{"key":"ref2","doi-asserted-by":"publisher","unstructured":"Samali A, Orrenius S. Heat shock proteins: regulators of stress response and apoptosis. Cell Stress & Chaperones. 1998; 3: 228\u2013236. <0228:hspros>2.3.co;2.","DOI":"10.1379\/1466-1268(1998)003"},{"key":"ref3","doi-asserted-by":"publisher","unstructured":"Fulda S, Gorman AM, Hori O, Samali A. Cellular stress responses: cell survival and cell death. International Journal of Cell Biology. 2010; 2010: 214074.","DOI":"10.1155\/2010\/214074"},{"key":"ref4","doi-asserted-by":"publisher","unstructured":"Fulda S. Evasion of apoptosis as a cellular stress response in cancer. International Journal of Cell Biology. 2010; 2010: 370835.","DOI":"10.1155\/2010\/370835"},{"key":"ref5","doi-asserted-by":"publisher","unstructured":"Perkins ND. The diverse and complex roles of NF-\u03baB subunits in cancer. Nature Reviews. Cancer. 2012; 12: 121\u2013132.","DOI":"10.1038\/nrc3204"},{"key":"ref6","doi-asserted-by":"publisher","unstructured":"Lingappan K. NF-\u03baB in Oxidative Stress. Current Opinion in Toxicology. 2018; 7: 81\u201386.","DOI":"10.1016\/j.cotox.2017.11.002"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"Clarkson RW, Heeley JL, Chapman R, Aillet F, Hay RT, Wyllie A, et al. NF-kappaB inhibits apoptosis in murine mammary epithelia. The Journal of Biological Chemistry. 2000; 275: 12737\u201312742.","DOI":"10.1074\/jbc.275.17.12737"},{"key":"ref8","doi-asserted-by":"publisher","unstructured":"Wang W, Nag SA, Zhang R. Targeting the NF\u03baB signaling pathways for breast cancer prevention and therapy. Current Medicinal Chemistry. 2015; 22: 264\u2013289.","DOI":"10.2174\/0929867321666141106124315"},{"key":"ref9","doi-asserted-by":"publisher","unstructured":"Sharif O, Bolshakov VN, Raines S, Newham P, Perkins ND. Transcriptional profiling of the LPS induced NF-kappaB response in macrophages. BMC Immunology. 2007; 8: 1.","DOI":"10.1186\/1471-2172-8-1"},{"key":"ref10","doi-asserted-by":"publisher","unstructured":"Xu Y, Jagannath C, Liu XD, Sharafkhaneh A, Kolodziejska KE, Eissa NT. Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity. 2007; 27: 135\u2013144.","DOI":"10.1016\/j.immuni.2007.05.022"},{"key":"ref11","doi-asserted-by":"publisher","unstructured":"Park HS, Jung HY, Park EY, Kim J, Lee WJ, Bae YS. Cutting edge: direct interaction of TLR4 with NAD(P)H oxidase 4 isozyme is essential for lipopolysaccharide-induced production of reactive oxygen species and activation of NF-kappa B. Journal of Immunology (Baltimore, Md.: 1950). 2004; 173: 3589\u20133593.","DOI":"10.4049\/jimmunol.173.6.3589"},{"key":"ref12","doi-asserted-by":"publisher","unstructured":"Page MJ, Kell DB, Pretorius E. The Role of Lipopolysaccharide-Induced Cell Signalling in Chronic Inflammation. Chronic Stress (Thousand Oaks, Calif.). 2022; 6: 24705470221076390.","DOI":"10.1177\/24705470221076390"},{"key":"ref13","doi-asserted-by":"publisher","unstructured":"Meier-Soelch J, Mayr-Buro C, Juli J, Leib L, Linne U, Dreute J, et al. Monitoring the Levels of Cellular NF-\u03baB Activation States. Cancers. 2021; 13: 5351.","DOI":"10.3390\/cancers13215351"},{"key":"ref14","doi-asserted-by":"crossref","unstructured":"Wookey PJ, Zulli A, Lo C, Hare DL, Schwarer AP, Darby IA, et al. Calcitonin receptor expression in embryonic, foetal and adult tissues: developmental and pathophysiological implications. In The calcitonin gene-related peptide family: form, function and future perspectives (pp. 199\u2013233). Springer: Berlin, Germany. 2009.","DOI":"10.1007\/978-90-481-2909-6_13"},{"key":"ref15","doi-asserted-by":"publisher","unstructured":"Pondel M. Calcitonin and calcitonin receptors: bone and beyond. International Journal of Experimental Pathology. 2000; 81: 405\u2013422.","DOI":"10.1046\/j.1365-2613.2000.00176.x"},{"key":"ref16","doi-asserted-by":"publisher","unstructured":"Furness S, Hare DL, Kourakis A, Turnley AM, Wookey PJ. A novel ligand of calcitonin receptor reveals a potential new sensor that modulates programmed cell death. Cell Death Discovery. 2016; 2: 16062.","DOI":"10.1038\/cddiscovery.2016.16062"},{"key":"ref17","doi-asserted-by":"publisher","unstructured":"Plotkin LI, Weinstein RS, Parfitt AM, Roberson PK, Manolagas SC, Bellido T. Prevention of osteocyte and osteoblast apoptosis by bisphosphonates and calcitonin. The Journal of Clinical Investigation. 1999; 104: 1363\u20131374.","DOI":"10.1172\/JCI6800"},{"key":"ref18","doi-asserted-by":"publisher","unstructured":"Findlay DM, Raggatt LJ, Bouralexis S, Hay S, Atkins GJ, Evdokiou A. Calcitonin decreases the adherence and survival of HEK-293 cells by a caspase-independent mechanism. The Journal of Endocrinology. 2002; 175: 715\u2013725.","DOI":"10.1677\/joe.0.1750715"},{"key":"ref19","doi-asserted-by":"publisher","unstructured":"Selander KS, H\u00e4rk\u00f6nen PL, Valve E, M\u00f6nkk\u00f6nen J, Hannuniemi R, V\u00e4\u00e4n\u00e4nen HK. Calcitonin promotes osteoclast survival in vitro. Molecular and Cellular Endocrinology. 1996; 122: 119\u2013129.","DOI":"10.1016\/0303-7207(96)03870-1"},{"key":"ref20","doi-asserted-by":"crossref","unstructured":"Han B, Nakamura M, Zhou G, Ishii A, Nakamura A, Bai Y, et al. Calcitonin inhibits invasion of breast cancer cells: involvement of urokinase-type plasminogen activator (uPA) and uPA receptor. International Journal of Oncology. 2006; 28: 807\u2013814.","DOI":"10.3892\/ijo.28.4.807"},{"key":"ref21","doi-asserted-by":"crossref","unstructured":"Thomas S, Muralidharan A, Shah GV. Knock-down of calcitonin receptor expression induces apoptosis and growth arrest of prostate cancer cells. International Journal of Oncology. 2007; 31: 1425\u20131437.","DOI":"10.3892\/ijo.31.6.1425"},{"key":"ref22","doi-asserted-by":"publisher","unstructured":"He T, Ling F. CALCR knockdown inhibits the development and progression of non-small-cell lung cancer. Carcinogenesis. 2021; 42: 1390\u20131398.","DOI":"10.1093\/carcin\/bgab076"},{"key":"ref23","doi-asserted-by":"publisher","unstructured":"Wang X, Nakamura M, Mori I, Takeda K, Nakamura Y, Utsunomiya H, et al. Calcitonin receptor gene and breast cancer: quantitative analysis with laser capture microdissection. Breast Cancer Research and Treatment. 2004; 83: 109\u2013117.","DOI":"10.1023\/B:BREA.0000010703.59483.c0"},{"key":"ref24","doi-asserted-by":"publisher","unstructured":"Gillespie MT, Thomas RJ, Pu ZY, Zhou H, Martin TJ, Findlay DM. Calcitonin receptors, bone sialoprotein and osteopontin are expressed in primary breast cancers. International Journal of Cancer. 1997; 73: 812\u2013815. <812::aid-ijc7>3.0.co;2-5.","DOI":"10.1002\/(sici)1097-0215(19971210)73:6"},{"key":"ref25","doi-asserted-by":"publisher","unstructured":"Siegl D, Kruchem M, Jansky S, Eichler E, Thies D, Hartwig U, et al. A PCR protocol to establish standards for routine mycoplasma testing that by design detects over ninety percent of all known mycoplasma species. iScience. 2023; 26: 106724.","DOI":"10.1016\/j.isci.2023.106724"},{"key":"ref26","doi-asserted-by":"publisher","unstructured":"Liu H, Wang L, Wang X, Cao Z, Yang Q, Zhang K. S100A7 enhances invasion of human breast cancer MDA-MB-468 cells through activation of nuclear factor-\u03baB signaling. World Journal of Surgical Oncology. 2013; 11: 93.","DOI":"10.1186\/1477-7819-11-93"},{"key":"ref27","doi-asserted-by":"publisher","unstructured":"Koh JY, Wie MB, Gwag BJ, Sensi SL, Canzoniero LM, Demaro J, et al. Staurosporine-induced neuronal apoptosis. Experimental Neurology. 1995; 135: 153\u2013159.","DOI":"10.1006\/exnr.1995.1074"},{"key":"ref28","doi-asserted-by":"publisher","unstructured":"Qin H, Wilson CA, Lee SJ, Zhao X, Benveniste EN. LPS induces CD40 gene expression through the activation of NF-kappaB and STAT-1alpha in macrophages and microglia. Blood. 2005; 106: 3114\u20133122.","DOI":"10.1182\/blood-2005-02-0759"},{"key":"ref29","unstructured":"Cordelieres FP, Bolte S. JACoP v2. 0: improving the user experience with co-localization studies. In ImageJ User & Developer Conference. 2008."},{"key":"ref30","doi-asserted-by":"crossref","unstructured":"Ziegler-Heitbrock HW, Wedel A, Schraut W, Str\u00f6bel M, Wendelgass P, Sternsdorf T, et al. Tolerance to lipopolysaccharide involves mobilization of nuclear factor kappa B with predominance of p50 homodimers. The Journal of Biological Chemistry. 1994; 269: 17001\u201317004.","DOI":"10.1016\/S0021-9258(17)32510-3"},{"key":"ref31","doi-asserted-by":"publisher","unstructured":"Skurski J, Dixit G, Blobel CP, Issuree PD, Maretzky T. The Threshold Effect: Lipopolysaccharide-Induced Inflammatory Responses in Primary Macrophages Are Differentially Regulated in an iRhom2-Dependent Manner. Frontiers in Cellular and Infection Microbiology. 2021; 10: 620392.","DOI":"10.3389\/fcimb.2020.620392"},{"key":"ref32","doi-asserted-by":"publisher","unstructured":"Ciccotosto GD, Jana M, Galluzzi L, Mitchell L, Gupta P, Furness SG, et al. A novel pH-sensitive probe to quantify autophagy on high throughput\/content imaging platforms. BioRxiv. 2025.  (preprint)","DOI":"10.1101\/2025.05.13.653902"},{"key":"ref33","doi-asserted-by":"publisher","unstructured":"Liu Y, He J, Chen J, Chen T, Li W, Yang Z, et al. Programmed cell death in triple-negative breast cancer. Cellular & Molecular Biology Letters. 2025; 30: 111.","DOI":"10.1186\/s11658-025-00789-5"},{"key":"ref34","doi-asserted-by":"publisher","unstructured":"Beaudreuil J, Balasubramanian S, Chenais J, Taboulet J, Frenkian M, Orcel P, et al. Molecular characterization of two novel isoforms of the human calcitonin receptor. Gene. 2004; 343: 143\u2013151.","DOI":"10.1016\/j.gene.2004.08.019"},{"key":"ref35","doi-asserted-by":"publisher","unstructured":"Albrandt K, Brady EM, Moore CX, Mull E, Sierzega ME, Beaumont K. Molecular cloning and functional expression of a third isoform of the human calcitonin receptor and partial characterization of the calcitonin receptor gene. Endocrinology. 1995; 136: 5377\u20135384.","DOI":"10.1210\/endo.136.12.7588285"},{"key":"ref36","doi-asserted-by":"publisher","unstructured":"K\u00f6rner M, Miller LJ. Alternative splicing of pre-mRNA in cancer: focus on G protein-coupled peptide hormone receptors. The American Journal of Pathology. 2009; 175: 461\u2013472.","DOI":"10.2353\/ajpath.2009.081135"},{"key":"ref37","doi-asserted-by":"publisher","unstructured":"Bar-Shavit R, Maoz M, Kancharla A, Nag JK, Agranovich D, Grisaru-Granovsky S, et al. G Protein-Coupled Receptors in Cancer. International Journal of Molecular Sciences. 2016; 17: 1320.","DOI":"10.3390\/ijms17081320"},{"key":"ref38","doi-asserted-by":"crossref","unstructured":"Wookey PJ, Gupta P, Hare DL, Ciccotosto GD, Hutchings CJ. Splice Variants of G Protein-Coupled Receptors Expressed in Cancers: Effective Targeting with Monoclonal Antibodies and Antibody-Like Scaffolds As Ligands Irrespective of the Pharmacological Status of Isoforms. In Handbook of Cancer and Immunology (pp. 1\u201345). Springer: Cham. 2022.","DOI":"10.1007\/978-3-030-80962-1_261-1"},{"key":"ref39","doi-asserted-by":"publisher","unstructured":"Moore EE, Kuestner RE, Stroop SD, Grant FJ, Matthewes SL, Brady CL, et al. Functionally different isoforms of the human calcitonin receptor result from alternative splicing of the gene transcript. Molecular Endocrinology (Baltimore, Md.). 1995; 9: 959\u2013968.","DOI":"10.1210\/mend.9.8.7476993"},{"key":"ref40","doi-asserted-by":"publisher","unstructured":"Ahn KS, Aggarwal BB. Transcription factor NF-kappaB: a sensor for smoke and stress signals. Annals of the New York Academy of Sciences. 2005; 1056: 218\u2013233.","DOI":"10.1196\/annals.1352.026"},{"key":"ref41","doi-asserted-by":"publisher","unstructured":"Kassem A, Henning P, Kindlund B, Lindholm C, Lerner UH. TLR5, a novel mediator of innate immunity-induced osteoclastogenesis and bone loss. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology. 2015; 29: 4449\u20134460.","DOI":"10.1096\/fj.15-272559"},{"key":"ref42","doi-asserted-by":"publisher","unstructured":"Miller SC, Huang R, Sakamuru S, Shukla SJ, Attene-Ramos MS, Shinn P, et al. Identification of known drugs that act as inhibitors of NF-kappaB signaling and their mechanism of action. Biochemical Pharmacology. 2010; 79: 1272\u20131280.","DOI":"10.1016\/j.bcp.2009.12.021"},{"key":"ref43","doi-asserted-by":"publisher","unstructured":"Tsao P, Cao T, von Zastrow M. Role of endocytosis in mediating downregulation of G-protein-coupled receptors. Trends in Pharmacological Sciences. 2001; 22: 91\u201396.","DOI":"10.1016\/s0165-6147(00)01620-5"},{"key":"ref44","doi-asserted-by":"publisher","unstructured":"Seck T, Baron R, Horne WC. Binding of filamin to the C-terminal tail of the calcitonin receptor controls recycling. The Journal of Biological Chemistry. 2003; 278: 10408\u201310416.","DOI":"10.1074\/jbc.M209655200"},{"key":"ref45","doi-asserted-by":"publisher","unstructured":"Chambers TJ. Osteoblasts release osteoclasts from calcitonin-induced quiescence. Journal of Cell Science. 1982; 57: 247\u2013260.","DOI":"10.1242\/jcs.57.1.247"},{"key":"ref46","unstructured":"Ng KW, Livesey SA, Larkins RG, Martin TJ. Calcitonin effects on growth and on selective activation of type II isoenzyme of cyclic adenosine 3\u2019:5\u2019-monophosphate-dependent protein kinase in T 47D human breast cancer cells. Cancer Research. 1983; 43: 794\u2013800."},{"key":"ref47","doi-asserted-by":"publisher","unstructured":"Lacroix M, Siwek B, Body JJ. Breast cancer cell response to calcitonin: modulation by growth-regulating agents. European Journal of Pharmacology. 1998; 344: 279\u2013286.","DOI":"10.1016\/s0014-2999(97)01578-1"},{"key":"ref48","doi-asserted-by":"publisher","unstructured":"Wang S, Wang W, Zeng J. Role of CALCR expression in liver cancer: Implications for the immunotherapy response. Molecular Medicine Reports. 2025; 31: 41.","DOI":"10.3892\/mmr.2024.13406"},{"key":"ref49","doi-asserted-by":"publisher","unstructured":"Alberti G, S\u00e1nchez-L\u00f3pez CM, Marcilla A, Barone R, Caruso Bavisotto C, Graziano F, et al. Hsp70 and Calcitonin Receptor Protein in Extracellular Vesicles from Glioblastoma Multiforme: Biomarkers with Putative Roles in Carcinogenesis and Potential for Differentiating Tumor Types. International Journal of Molecular Sciences. 2024; 25: 3415.","DOI":"10.3390\/ijms25063415"},{"key":"ref50","doi-asserted-by":"publisher","unstructured":"Hay DL, Garelja ML, Poyner DR, Walker CS. Update on the pharmacology of calcitonin\/CGRP family of peptides: IUPHAR Review 25. British Journal of Pharmacology. 2018; 175: 3\u201317.","DOI":"10.1111\/bph.14075"}],"container-title":["Frontiers in Bioscience-Landmark"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.imrpress.com\/journal\/FBL\/31\/4\/10.31083\/FBL50148","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T09:29:47Z","timestamp":1777454987000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.imrpress.com\/journal\/FBL\/31\/4\/10.31083\/FBL50148"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,24]]},"references-count":50,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2026]]}},"alternative-id":["S2768-6701(26)02071-X"],"URL":"https:\/\/doi.org\/10.31083\/fbl50148","relation":{},"ISSN":["2768-6701","2768-6698"],"issn-type":[{"value":"2768-6701","type":"print"},{"value":"2768-6698","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,24]]},"article-number":"50148"}}