{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T06:53:50Z","timestamp":1770533630695,"version":"3.49.0"},"reference-count":43,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,5,30]],"date-time":"2024-05-30T00:00:00Z","timestamp":1717027200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Basic Science Research Program through the National Research Foundation of Korea (NRF)","award":["NRF-2018R1A6A1A03023718"],"award-info":[{"award-number":["NRF-2018R1A6A1A03023718"]}]},{"DOI":"10.13039\/501100003725","name":"Ministry of Education","doi-asserted-by":"publisher","award":["NRF-2018R1A6A1A03023718"],"award-info":[{"award-number":["NRF-2018R1A6A1A03023718"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Anoctamin1 (ANO1), a calcium-activated chloride channel, is overexpressed in a variety of cancer cells, including prostate cancer, and is involved in cancer cell proliferation, migration, and invasion. Inhibition of ANO1 in these cancer cells exhibits anticancer effects. In this study, we conducted a screening to identify novel ANO1 inhibitors with anticancer effects using PC-3 human prostate carcinoma cells. Screening of 2978 approved and investigational drugs revealed that hemin is a novel ANO1 inhibitor with an IC50 value of 0.45 \u03bcM. Notably, hemin had no significant effect on intracellular calcium signaling and cystic fibrosis transmembrane conductance regulator (CFTR), a cyclic AMP (cAMP)-regulated chloride channel, and it showed a weak inhibitory effect on ANO2 at 3 \u03bcM, a concentration that completely inhibits ANO1. Interestingly, hemin also significantly decreased ANO1 protein levels and strongly inhibited the cell proliferation and migration of PC-3 cells in an ANO1-dependent manner. Furthermore, it strongly induced caspase-3 activation, PARP degradation, and apoptosis in PC-3 cells. These findings suggest that hemin possesses anticancer properties via ANO1 inhibition and could be considered for development as a novel treatment for prostate cancer.<\/jats:p>","DOI":"10.3390\/ijms25116032","type":"journal-article","created":{"date-parts":[[2024,5,31]],"date-time":"2024-05-31T03:46:49Z","timestamp":1717127209000},"page":"6032","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Anticancer Effect of Hemin through ANO1 Inhibition in Human Prostate Cancer Cells"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0414-6245","authenticated-orcid":false,"given":"So-Hyeon","family":"Park","sequence":"first","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3224-6510","authenticated-orcid":false,"given":"Yechan","family":"Lee","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0711-0766","authenticated-orcid":false,"given":"Hyejin","family":"Jeon","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junghwan","family":"Park","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jieun","family":"Kim","sequence":"additional","affiliation":[{"name":"Graduate Program of Industrial Pharmaceutical Science, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mincheol","family":"Kang","sequence":"additional","affiliation":[{"name":"Graduate Program of Industrial Pharmaceutical Science, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1480-7433","authenticated-orcid":false,"given":"Wan","family":"Namkung","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"},{"name":"Graduate Program of Industrial Pharmaceutical Science, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1126\/science.1163518","article-title":"TMEM16A, a membrane protein associated with calcium-dependent chloride channel activity","volume":"322","author":"Caputo","year":"2008","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1016\/j.cell.2008.09.003","article-title":"Expression cloning of TMEM16A as a calcium-activated chloride channel subunit","volume":"134","author":"Schroeder","year":"2008","journal-title":"Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1210","DOI":"10.1038\/nature07313","article-title":"TMEM16A confers receptor-activated calcium-dependent chloride conductance","volume":"455","author":"Yang","year":"2008","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"21413","DOI":"10.1073\/pnas.0911935106","article-title":"Studies on expression and function of the TMEM16A calcium-activated chloride channel","volume":"106","author":"Huang","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4887","DOI":"10.1113\/jphysiol.2009.176198","article-title":"Expression of anoctamin 1\/TMEM16A by interstitial cells of Cajal is fundamental for slow wave activity in gastrointestinal muscles","volume":"587","author":"Hwang","year":"2009","journal-title":"J. Physiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1152\/physiol.00030.2010","article-title":"TMEM16A protein: A new identity for Ca2+-dependent Cl\u2212 channels","volume":"25","author":"Ferrera","year":"2010","journal-title":"Physiology"},{"key":"ref_7","first-page":"413","article-title":"Chromosome 11q13 abnormalities in human cancer","volume":"3","author":"Lammie","year":"1991","journal-title":"Cancer Cells"},{"key":"ref_8","first-page":"5229","article-title":"Amplification of genes within the chromosome 11q13 region is indicative of poor prognosis in patients with operable breast cancer","volume":"52","author":"Schuuring","year":"1992","journal-title":"Cancer Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1068","DOI":"10.3892\/mmr.2014.1888","article-title":"Transmembrane protein with unknown function 16A overexpression promotes glioma formation through the nuclear factor-\u03baB signaling pathway","volume":"9","author":"Liu","year":"2014","journal-title":"Mol. Med. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3270","DOI":"10.1158\/0008-5472.CAN-12-0475-T","article-title":"TMEM16A induces MAPK and contributes directly to tumorigenesis and cancer progression","volume":"72","author":"Duvvuri","year":"2012","journal-title":"Cancer Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/S0002-9440(10)63279-8","article-title":"The novel marker, DOG1, is expressed ubiquitously in gastrointestinal stromal tumors irrespective of KIT or PDGFRA mutation status","volume":"165","author":"West","year":"2004","journal-title":"Am. J. Pathol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3297","DOI":"10.1007\/s11033-023-08293-9","article-title":"High level of ANO1 promotes pancreatic cancer growth in concert with oncogenic KRAS","volume":"50","author":"Zhang","year":"2023","journal-title":"Mol. Biol. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/nature13176","article-title":"Identification of genomic alterations in oesophageal squamous cell cancer","volume":"509","author":"Song","year":"2014","journal-title":"Nature"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"E1026","DOI":"10.1073\/pnas.1217072110","article-title":"Calcium-activated chloride channel ANO1 promotes breast cancer progression by activating EGFR and CAMK signaling","volume":"110","author":"Britschgi","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"9722","DOI":"10.1073\/pnas.1423827112","article-title":"Anoctamin 1 (TMEM16A) is essential for testosterone-induced prostate hyperplasia","volume":"112","author":"Cha","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jia, L., Liu, W., Guan, L., Lu, M., and Wang, K. (2015). Inhibition of Calcium-Activated Chloride Channel ANO1\/TMEM16A Suppresses Tumor Growth and Invasion in Human Lung Cancer. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0136584"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.canlet.2012.07.015","article-title":"Inhibition of Ca2+-activated Cl\u2212 channel ANO1\/TMEM16A expression suppresses tumor growth and invasiveness in human prostate carcinoma","volume":"326","author":"Liu","year":"2012","journal-title":"Cancer Lett."},{"key":"ref_18","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_19","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.eururo.2019.08.005","article-title":"Recent Global Patterns in Prostate Cancer Incidence and Mortality Rates","volume":"77","author":"Culp","year":"2020","journal-title":"Eur. Urol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1124\/mol.113.087502","article-title":"MONNA, a potent and selective blocker for transmembrane protein with unknown function 16\/anoctamin-1","volume":"84","author":"Oh","year":"2013","journal-title":"Mol. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1124\/mol.107.043208","article-title":"Small-molecule screen identifies inhibitors of a human intestinal calcium-activated chloride channel","volume":"73","author":"Namkung","year":"2008","journal-title":"Mol. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Seo, Y., Park, J., Kim, M., Lee, H.K., Kim, J.H., Jeong, J.H., and Namkung, W. (2015). Inhibition of ANO1\/TMEM16A Chloride Channel by Idebenone and Its Cytotoxicity to Cancer Cell Lines. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0133656"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.ejmech.2018.10.002","article-title":"Synthesis and biological evaluation of novel Ani9 derivatives as potent and selective ANO1 inhibitors","volume":"160","author":"Seo","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Seo, Y., Lee, H.K., Park, J., Jeon, D.K., Jo, S., Jo, M., and Namkung, W. (2016). Ani9, A Novel Potent Small-Molecule ANO1 Inhibitor with Negligible Effect on ANO2. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0155771"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Seo, Y., Anh, N.H., Heo, Y., Park, S.H., Kiem, P.V., Lee, Y., Yen, D.T.H., Jo, S., Jeon, D., and Tai, B.H. (2020). Novel ANO1 Inhibitor from Mallotus apelta Extract Exerts Anticancer Activity through Downregulation of ANO1. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21186470"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Seo, Y., Ryu, K., Park, J., Jeon, D.K., Jo, S., Lee, H.K., and Namkung, W. (2017). Inhibition of ANO1 by luteolin and its cytotoxicity in human prostate cancer PC-3 cells. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0174935"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"439","DOI":"10.7326\/0003-4819-142-6-200503150-00010","article-title":"Recommendations for the diagnosis and treatment of the acute porphyrias","volume":"142","author":"Anderson","year":"2005","journal-title":"Ann. Intern. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"16354","DOI":"10.1073\/pnas.1214596109","article-title":"Calcium-activated chloride channel TMEM16A modulates mucin secretion and airway smooth muscle contraction","volume":"109","author":"Huang","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1038\/s41419-018-0735-2","article-title":"Inhibition of ANO1\/TMEM16A induces apoptosis in human prostate carcinoma cells by activating TNF-\u03b1 signaling","volume":"9","author":"Song","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_30","unstructured":"Dirckx, J.H. (2001). Stedman\u2019s Concise Medical Dictionary for the Health Professions, Lippincott Williams & Wilkins. [4th ed.]."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"17","DOI":"10.14218\/JCTH.2014.00039","article-title":"Acute Hepatic Porphyria","volume":"3","author":"Bissell","year":"2015","journal-title":"J. Clin. Transl. Hepatol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"801.e1","DOI":"10.1016\/j.amjmed.2006.05.026","article-title":"Open-label study of hemin for acute porphyria: Clinical practice implications","volume":"119","author":"Anderson","year":"2006","journal-title":"Am. J. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1821","DOI":"10.1038\/sj.onc.1209203","article-title":"Head and neck squamous cell carcinoma transcriptome analysis by comprehensive validated differential display","volume":"25","author":"Carles","year":"2006","journal-title":"Oncogene"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"10091","DOI":"10.1038\/s41598-023-35125-4","article-title":"Proteomic analysis of the effect of hemin in breast cancer","volume":"13","author":"Schweitzer","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5135","DOI":"10.1158\/1078-0432.CCR-17-0112","article-title":"In Vivo Hemin Conditioning Targets the Vascular and Immunologic Compartments and Restrains Prostate Tumor Development","volume":"23","author":"Jaworski","year":"2017","journal-title":"Clin. Cancer Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2727","DOI":"10.1002\/2211-5463.13271","article-title":"Hemin activation abrogates Mycoplasma hyorhinis replication in chronically infected prostate cancer cells via heme oxygenase-1 induction","volume":"11","author":"Huang","year":"2021","journal-title":"FEBS Open Bio"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"112946","DOI":"10.1016\/j.yexcr.2021.112946","article-title":"Hemin enhances radiosensitivity of lung cancer cells through ferroptosis","volume":"410","author":"Almahi","year":"2022","journal-title":"Exp. Cell Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"153530","DOI":"10.1016\/j.prp.2021.153530","article-title":"Hemin, a major heme molecule, induced cellular and genetic alterations in normal colonic and colon cancer cells","volume":"224","author":"Cheng","year":"2021","journal-title":"Pathol. Res. Pract."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2030","DOI":"10.1089\/ars.2018.7554","article-title":"Heme Oxygenase-1 Has an Antitumor Role in Breast Cancer","volume":"30","author":"Gandini","year":"2019","journal-title":"Antioxid. Redox Signal"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Nitti, M., Piras, S., Marinari, U.M., Moretta, L., Pronzato, M.A., and Furfaro, A.L. (2017). HO-1 Induction in Cancer Progression: A Matter of Cell Adaptation. Antioxidants, 6.","DOI":"10.3390\/antiox6020029"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1745","DOI":"10.1158\/1541-7786.MCR-08-0325","article-title":"Critical role of endogenous heme oxygenase 1 as a tuner of the invasive potential of prostate cancer cells","volume":"7","author":"Gueron","year":"2009","journal-title":"Mol. Cancer Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4301","DOI":"10.3390\/cimb45050273","article-title":"Pharmacological Significance of Heme Oxygenase 1 in Prostate Cancer","volume":"45","author":"Gayle","year":"2023","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_43","first-page":"e2597","article-title":"Modified annexin V\/propidium iodide apoptosis assay for accurate assessment of cell death","volume":"50","author":"Rieger","year":"2011","journal-title":"J. Vis. Exp."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/11\/6032\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:51:18Z","timestamp":1760107878000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/11\/6032"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,30]]},"references-count":43,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["ijms25116032"],"URL":"https:\/\/doi.org\/10.3390\/ijms25116032","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,30]]}}}