{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T07:28:56Z","timestamp":1787902136213,"version":"build-2784847793"},"reference-count":34,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,3,31]],"date-time":"2021-03-31T00:00:00Z","timestamp":1617148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Antitumor effects of shikonins on chronic lymphocytic leukemia (CLL) and B-cell prolymphocytic leukemia (B-PLL) are mostly unexplored. The antitumor activity of shikonins, isolated from Onosma visianii Clem (Boraginaceae), in BCL1, mouse CLL cells and JVM-13, human B-PLL cells was explored in this study. The cytotoxicity of shikonin derivatives was measured by an MTT test. Cell death, proliferation, cell cycle, and expression of molecules that control these processes were analyzed by flow cytometry. Expression of STAT3-regulated genes was analyzed by real-time q-RT-PCR (Quantitative Real-Time Polymerase Chain Reaction). The antitumor effects of shikonin derivatives in vivo were analyzed, using flow cytometry, by detection of leukemia cells in the peripheral blood and spleens of mice intravenously injected with BCL1 cells. The two most potent derivatives, isobutyrylshikonin (IBS) and \u03b1-methylbutyrylshikonin (MBS), induced cell cycle disturbances and apoptosis, inhibited proliferation, and decreased expression of phospho-STAT3 and downstream-regulated molecules in BCL1 and JVM-13 cells. IBS and MBS decreased the percentage of leukemia cells in vivo. The link between the decrease in phosphorylated STAT3 by MBS and IBS and BCL1 cell death was confirmed by detection of enhanced cell death after addition of AG490, an inhibitor of Jak2 kinase. It seems that IBS and MBS, by decreasing STAT3 phosphorylation, trigger apoptosis, inhibit cell proliferation, and attenuate leukemia cell stemness.<\/jats:p>","DOI":"10.3390\/nu13041147","type":"journal-article","created":{"date-parts":[[2021,3,31]],"date-time":"2021-03-31T05:57:18Z","timestamp":1617170238000},"page":"1147","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Shikonin Derivatives from Onsoma visianii Decrease Expression of Phosphorylated STAT3 in Leukemia Cells and Exert Antitumor Activity"],"prefix":"10.3390","volume":"13","author":[{"given":"Zeljko","family":"Todorovic","sequence":"first","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jelena","family":"Milovanovic","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine and Stem Cell Research, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"},{"name":"Department of Histology and Embriology, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5357-2633","authenticated-orcid":false,"given":"Dragana","family":"Arsenijevic","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine and Stem Cell Research, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"},{"name":"Department of Pharmacy, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nenad","family":"Vukovic","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Science, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7222-7245","authenticated-orcid":false,"given":"Milena","family":"Vukic","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Science, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandar","family":"Arsenijevic","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine and Stem Cell Research, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Predrag","family":"Djurdjevic","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marija","family":"Milovanovic","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine and Stem Cell Research, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nebojsa","family":"Arsenijevic","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine and Stem Cell Research, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1685","DOI":"10.1055\/s-0033-1350934","article-title":"Pharmacological properties of shikonin\u2014A review of literature since 2002","volume":"79","author":"Giner","year":"2013","journal-title":"Planta Med."},{"key":"ref_2","first-page":"73","article-title":"Antibacterial and cytotoxic activities of naphthoquinone pigments from Onosma visianii Clem","volume":"16","author":"Vukic","year":"2017","journal-title":"EXCLI J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"818709","DOI":"10.1155\/2013\/818709","article-title":"Integration of Different \u201c-omics\u201d Technologies Identifies Inhibition of the IGF1R-Akt-mTOR Signaling Cascade Involved in the Cytotoxic Effect of Shikonin against Leukemia Cells","volume":"2013","author":"Wiench","year":"2013","journal-title":"Evid. Based Complementary Altern. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1124\/mi.11.1.4","article-title":"STAT3 signaling: Anticancer strategies and challenges","volume":"11","author":"Johnston","year":"2011","journal-title":"Mol. Interv."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2852","DOI":"10.1182\/blood-2009-10-230060","article-title":"STAT3 is constitutively phosphorylated on serine 727 residues, binds DNA, and activates transcription in CLL cells","volume":"115","author":"Harris","year":"2010","journal-title":"Blood"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3853409","DOI":"10.1155\/2018\/3853409","article-title":"Proscillaridin A Promotes Oxidative Stress and ER Stress, Inhibits STAT3 Activation, and Induces Apoptosis in A549 Lung Adenocarcinoma Cells","volume":"2018","author":"Maryam","year":"2018","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.5115\/acb.2014.47.1.1","article-title":"The expressions of stem cell markers: Oct4, Nanog, Sox2, nucleostemin, Bmi, Zfx, Tcl1, Tbx3, Dppa4, and Esrrb in bladder, colon, and prostate cancer, and certain cancer cell lines","volume":"47","author":"Amini","year":"2014","journal-title":"Anat. Cell Biol."},{"key":"ref_8","unstructured":"Noone, A.M., Howlader, N., Krapcho, M., Miller, D., Brest, A., Yu, M., Ruhl, J., Tatalovich, Z., Mariotto, A., and Lewis, D.R. (2018). SEER Cancer Statistics Review, 1975\u20132015, National Cancer Institute. Based on November 2017 SEER Data Submission, Posted to the SEER Web Site."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1056\/NEJMoa1513257","article-title":"Targeting BCL2 with venetoclax inrelapsed chronic lymphocytic leukemia","volume":"374","author":"Roberts","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2076","DOI":"10.1002\/ijc.30892","article-title":"Activation of the B-cell receptor successively activates NF-\u03baB and STAT3 in chronic lymphocytic leukemia cells","volume":"141","author":"Rozovski","year":"2017","journal-title":"Int. J. Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5019","DOI":"10.1182\/blood-2011-01-293050","article-title":"The 2008 WHO classification of lymphoid neoplasms and beyond: Evolving concepts and practical applications","volume":"117","author":"Campo","year":"2011","journal-title":"Blood"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1111\/j.1365-2141.1974.tb06769.x","article-title":"Prolymphocytic leukaemia","volume":"27","author":"Galton","year":"1974","journal-title":"Br. J. Haematol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1182\/blood-2012-01-380139","article-title":"How I treat prolymphocytic leukemia","volume":"120","author":"Dearden","year":"2012","journal-title":"Blood"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1007\/s10565-010-9167-1","article-title":"Haemotoxicity of busulphan, doxorubicin, cisplatin and cyclophosphamide in the female BALB\/c mouse using a brief regimen of drug administration","volume":"27","author":"Molyneux","year":"2011","journal-title":"Cell Biol. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Severin, F., Frezzato, F., Visentin, A., Martini, V., Trimarco, V., Carraro, S., Tibaldi, E., Brunati, A.M., Piazza, F., and Semenzato, G. (2019). In Chronic Lymphocytic Leukemia the JAK2\/STAT3 Pathway Is Constitutively Activated and Its Inhibition Leads to CLL Cell Death Unaffected by the Protective Bone Marrow Microenvironment. Cancers, 11.","DOI":"10.3390\/cancers11121939"},{"key":"ref_16","first-page":"339","article-title":"Stat3\/Oct-4\/c-Myc signal circuit for regulating stemness-mediated doxorubicin resistance of triple-negative breast cancer cells and inhibitory effects of WP1066","volume":"53","author":"Cheng","year":"2018","journal-title":"Int. J. Oncol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"10194","DOI":"10.1038\/srep10194","article-title":"Inhibition of STAT3, FAK and Src mediated signaling reduces cancer stem cell load, tumorigenic potential and metastasis in breast cancer","volume":"5","author":"Thakur","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1172\/JCI23409","article-title":"In vivo measurements document the dynamic cellular kinetics of chronic lymphocytic leukemia B cells","volume":"115","author":"Messmer","year":"2005","journal-title":"J. Clin. Investig."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1038\/cr.2008.86","article-title":"XInduction of apoptosis by shikonin through a ROS\/JNK-mediated process in Bcr\/Ablpositive chronic myelogenous leukemia (CML) cells","volume":"18","author":"Mao","year":"2008","journal-title":"Cell Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2712","DOI":"10.1080\/14786419.2017.1374271","article-title":"Naphthoquinone rich Onosma visianii Clem (Boraginaceae) root extracts induce apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines","volume":"32","author":"Vukic","year":"2018","journal-title":"Nat. Prod. Res."},{"key":"ref_21","first-page":"48","article-title":"Redox status, DNA and HSA binding study of naturally occurring naphthoquinone derivatives","volume":"19","author":"Vukic","year":"2020","journal-title":"EXCLI J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1159\/000490142","article-title":"The Critical Role of PTEN\/PI3K\/AKT Signaling Pathway in Shikonin-Induced Apoptosis and Proliferation Inhibition of Chronic Myeloid Leukemia","volume":"47","author":"Chen","year":"2018","journal-title":"Cell Physiol. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3055","DOI":"10.3892\/mmr.2017.6965","article-title":"Shikonin suppresses proliferation and induces apoptosis in human leukemia NB4 cells through modulation of MAPKs and c-Myc","volume":"16","author":"Shan","year":"2017","journal-title":"Mol. Med. Rep."},{"key":"ref_24","first-page":"3077","article-title":"Cisplatin-induced Apoptosis Proceeds by Caspase-3-dependent and -independent Pathways in Cisplatin-resistant and -sensitive Human Ovarian Cancer Cell Lines","volume":"59","author":"Henkels","year":"1999","journal-title":"Cancer Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1016\/j.immuni.2006.03.018","article-title":"The Noxa\/Mcl-1 axis regulates susceptibility to apoptosis under glucose limitation in dividing T cells","volume":"24","author":"Alves","year":"2006","journal-title":"Immunity"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s12929-015-0127-1","article-title":"Shikonin selectively induces apoptosis in human prostate cancer cells through the endoplasmic reticulum stress and mitochondrial apoptotic pathway","volume":"22","author":"Gara","year":"2015","journal-title":"J. Biomed. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2662","DOI":"10.1080\/21691401.2019.1632870","article-title":"Shikonin inhibits proliferation, migration, invasion and promotes apoptosis in NCI-N87 cells via inhibition of PI3K\/AKT signal pathway","volume":"47","author":"Jia","year":"2019","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2909","DOI":"10.1038\/sj.onc.1208618","article-title":"Cyclins and cdks in development and cancer: A perspective","volume":"24","author":"Deshpande","year":"2005","journal-title":"Oncogene"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.1101\/gad.13.12.1501","article-title":"CDK inhibitors: Positive and negative regulators of G1-phase progression","volume":"13","author":"Sherr","year":"1999","journal-title":"Genes Dev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1038\/366701a0","article-title":"p21 is a universal inhibitor of cyclin kinases","volume":"366","author":"Xiong","year":"1993","journal-title":"Nature"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1080\/13880209.2019.1626447","article-title":"Deoxyshikonin isolated from Arnebia euchroma inhibits colorectal cancer by down-regulating the PI3K\/Akt\/mTOR pathway","volume":"57","author":"Zhu","year":"2019","journal-title":"Pharm. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"13961","DOI":"10.1074\/jbc.274.20.13961","article-title":"A cyclin D-Cdk4 activity required for G2 phase cell cycle progression is inhibited in ultraviolet radiation-induced G2 phase delay","volume":"274","author":"Gabrielli","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1186\/s12935-018-0549-4","article-title":"Down-regulation of STAT3 induces the apoptosis and G1 cell cycle arrest in esophageal carcinoma ECA109 cells","volume":"18","author":"Zhou","year":"2018","journal-title":"Cancer Cell Int."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2131","DOI":"10.3892\/ol.2017.6454","article-title":"HSTAT3 influences the characteristics of stem cells in cervical carcinoma","volume":"14","author":"Wang","year":"2017","journal-title":"Oncol. Lett."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/4\/1147\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:55:45Z","timestamp":1760363745000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/4\/1147"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,31]]},"references-count":34,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["nu13041147"],"URL":"https:\/\/doi.org\/10.3390\/nu13041147","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,31]]}}}