{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T03:58:34Z","timestamp":1762055914419,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,6,22]],"date-time":"2022-06-22T00:00:00Z","timestamp":1655856000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Owens-OSF Professorship","award":["629859","337788","631222"],"award-info":[{"award-number":["629859","337788","631222"]}]},{"name":"Children\u2019s Hospital of Illinois","award":["629859","337788","631222"],"award-info":[{"award-number":["629859","337788","631222"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background: Gankyrin, a member of the 26S proteasome, is an overexpressed oncoprotein in hepatoblastoma (HBL) and hepatocellular carcinoma (HCC). Cjoc42 was the first small molecule inhibitor of Gankyrin developed; however, the IC50 values of &gt;50 \u03bcM made them unattractive for clinical use. Second-generation inhibitors demonstrate a stronger affinity toward Gankyrin and increased cytotoxicity. The aim of this study was to characterize the in vitro effects of three cjoc42 derivatives. Methods: Experiments were performed on the HepG2 (HBL) and Hep3B (pediatric HCC) cell lines. We evaluated the expression of TSPs, cell cycle markers, and stem cell markers by Western blotting and\/or real-time quantitative reverse transcription PCR. We also performed apoptotic, synergy, and methylation assays. Results: The treatment with cjoc42 derivatives led to an increase in TSPs and a dose-dependent decrease in the stem cell phenotype in both cell lines. An increase in apoptosis was only seen with AFM-1 and -2 in Hep3B cells. Drug synergy was seen with doxorubicin, and antagonism was seen with cisplatin. In the presence of cjoc42 derivatives, the 20S subunit of the 26S proteasome was more available to transport doxorubicin to the nucleus, leading to synergy. Conclusion: Small-molecule inhibitors for Gankyrin are a promising therapeutic strategy, especially in combination with doxorubicin.<\/jats:p>","DOI":"10.3390\/cancers14133068","type":"journal-article","created":{"date-parts":[[2022,6,22]],"date-time":"2022-06-22T21:31:06Z","timestamp":1655933466000},"page":"3068","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Second Generation Small Molecule Inhibitors of Gankyrin for the Treatment of Pediatric Liver Cancer"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0552-6947","authenticated-orcid":false,"given":"Amber M.","family":"D\u2019Souza","sequence":"first","affiliation":[{"name":"Department of Pediatrics, University of Illinois College of Medicine Peoria, 1 Illini Drive, Peoria, IL 61605, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manu","family":"Gnanamony","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of Illinois College of Medicine Peoria, 1 Illini Drive, Peoria, IL 61605, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria","family":"Thomas","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of Illinois College of Medicine Peoria, 1 Illini Drive, Peoria, IL 61605, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Hanley","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of Illinois College of Medicine Peoria, 1 Illini Drive, Peoria, IL 61605, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dipti","family":"Kanabar","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, St. John\u2019s University, 8000 Utopia Pkwy, Jamaica, NY 11439, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pedro","family":"de Alarcon","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of Illinois College of Medicine Peoria, 1 Illini Drive, Peoria, IL 61605, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0646-9964","authenticated-orcid":false,"given":"Aaron","family":"Muth","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, St. John\u2019s University, 8000 Utopia Pkwy, Jamaica, NY 11439, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolai","family":"Timchenko","sequence":"additional","affiliation":[{"name":"Division of Pediatric Surgery, Cincinnati Children\u2019s Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,22]]},"reference":[{"key":"ref_1","unstructured":"Howlader, N., Noone, A., Krapcho, M., Miller, D., Brest, A., Yu, M., Ruhl, J., Tatalovich, Z., Mariotto, A., and Lewis, D. (2022, June 01). SEER Cancer Statistics Review, 1975\u20132018, National Cancer Institute. Bethesda, MD, Based on November 2020 SEER Data Submission, Posted to the SEER Web Site, April 2021, Available online: https:\/\/seer.cancer.gov\/csr\/1975_2018\/."},{"key":"ref_2","unstructured":"Katzenstein, H.M. (2016, January 25\u201326). COG Clinical Trials for Children with Liver Cancer. Proceedings of the International Pediatric Liver Tumor Symposium, Houston, TX, USA."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1053\/j.sempedsurg.2016.09.002","article-title":"Malignant tumors of the liver in children","volume":"25","author":"Aronson","year":"2016","journal-title":"Semin. Pediatr. Surg."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2789","DOI":"10.1200\/JCO.2002.06.155","article-title":"Hepatocellular Carcinoma in Children and Adolescents: Results From the Pediatric Oncology Group and the Children\u2019s Cancer Group Intergroup Study","volume":"20","author":"Katzenstein","year":"2002","journal-title":"J. Clin. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2798","DOI":"10.1200\/JCO.2002.06.102","article-title":"Hepatocellular Carcinoma in Children: Results of the First Prospective Study of the International Society of Pediatric Oncology Group","volume":"20","author":"Czauderna","year":"2002","journal-title":"J. Clin. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"53","DOI":"10.12688\/f1000research.12239.1","article-title":"Hepatoblastoma: Current understanding, recent advances, and controversies","volume":"7","author":"Czauderna","year":"2018","journal-title":"F1000Research"},{"key":"ref_7","first-page":"36","article-title":"Present and Future Prospect of Small Molecule & Related Targeted Therapy Against Human Cancer","volume":"9","author":"Pathak","year":"2018","journal-title":"Vivechan Int. J. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/j.jcmgh.2018.05.007","article-title":"Gankyrin Promotes Tumor-Suppressor Protein Degradation to Drive Hepatocyte Proliferation","volume":"6","author":"Jiang","year":"2018","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1038\/s42003-018-0077-8","article-title":"PARP1 activation increases expression of modified tumor suppressors and pathways underlying development of aggressive hepatoblastoma","volume":"1","author":"Valanejad","year":"2018","journal-title":"Commun. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1002\/hep.29677","article-title":"C\/EBP\u03b1-dependent preneoplastic tumor foci are the origin of hepatocellular carcinoma and aggressive pediatric liver cancer","volume":"67","author":"Cast","year":"2017","journal-title":"Hepatology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1038\/71600","article-title":"Reduced stability of retinoblastoma protein by gankyrin, an oncogenic ankyrin-repeat protein overexpressed in hepatomas","volume":"6","author":"Higashitsuji","year":"2000","journal-title":"Nat. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1093\/carcin\/bgx050","article-title":"FXR-Gankyrin axis is involved in development of pediatric liver cancer","volume":"38","author":"Valanejad","year":"2017","journal-title":"Carcinogenesis"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"638","DOI":"10.3748\/wjg.v8.i4.638","article-title":"Overexpression of P28\/Gankyrin in Human Hepatocellular Carcinoma and Its Clinical Significance","volume":"8","author":"Fu","year":"2002","journal-title":"World J. Gastroenterol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1159\/000494969","article-title":"Gankyrin Contributes to Tumorigenesis and Chemoresistance in Sporadic Colorectal Cancer","volume":"100","author":"Sakurai","year":"2018","journal-title":"Digestion"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"81156","DOI":"10.18632\/oncotarget.13215","article-title":"Gankyrin sustains PI3K\/GSK-3\u03b2\/\u03b2-catenin signal activation and promotes colorectal cancer aggressiveness and progression","volume":"7","author":"He","year":"2016","journal-title":"Oncotarget"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1016\/j.prp.2015.09.010","article-title":"Clinicopathologic features and prognostic implications of Gankyrin protein expression in non-small cell lung cancer","volume":"211","author":"Wang","year":"2015","journal-title":"Pathol. Res. Pract."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"23732","DOI":"10.1038\/srep23732","article-title":"Discovery of a small-molecule binder of the oncoprotein gankyrin that modulates gankyrin activity in the cell","volume":"6","author":"Chattopadhyay","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"126889","DOI":"10.1016\/j.bmcl.2019.126889","article-title":"Structural modification of the aryl sulfonate ester of cjoc42 for enhanced gankyrin binding and anti-cancer activity","volume":"30","author":"Kanabar","year":"2019","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"127372","DOI":"10.1016\/j.bmcl.2020.127372","article-title":"Optimizing the aryl-triazole of cjoc42 for enhanced gankyrin binding and anti-cancer activity","volume":"30","author":"Kanabar","year":"2020","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2549","DOI":"10.1172\/JCI41933","article-title":"Elimination of C\/EBP\u03b1 through the ubiquitin-proteasome system promotes the development of liver cancer in mice","volume":"120","author":"Wang","year":"2010","journal-title":"J. Clin. Investig."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1002\/hep.26146","article-title":"Farnesoid X receptor inhibits gankyrin in mouse livers and prevents development of liver cancer","volume":"57","author":"Jiang","year":"2012","journal-title":"Hepatology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Suarez-Arnedo, A., Figueroa, F.T., Clavijo, C., Arbel\u00e1ez, P., Cruz, J.C., and Mu\u00f1oz-Camargo, C. (2020). An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays. PLoS ONE, 15.","DOI":"10.1101\/2020.04.20.050831"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/978-1-60327-017-5_2","article-title":"Flow Cytometry-Based Apoptosis Detection","volume":"559","author":"Wlodkowic","year":"2009","journal-title":"Apoptosis"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"12.8.1","DOI":"10.1002\/0471141755.ph1208s25","article-title":"Measurement of Cell Death in Mammalian Cells","volume":"25","author":"Cummings","year":"2004","journal-title":"Curr. Protoc. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2866","DOI":"10.1093\/bioinformatics\/btw230","article-title":"Combenefit: An interactive platform for the analysis and visualization of drug combinations","volume":"32","author":"Fornari","year":"2016","journal-title":"Bioinformatics"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1056\/NEJMoa0708857","article-title":"Sorafenib in Advanced Hepatocellular Carcinoma","volume":"359","author":"Llovet","year":"2008","journal-title":"N. Engl. J. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1080\/21691401.2017.1388250","article-title":"Gankyrin: A novel promising therapeutic target for hepatocellular carcinoma","volume":"46","author":"Zamani","year":"2018","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"15","DOI":"10.2165\/00003088-198815010-00002","article-title":"Clinical Pharmacokinetics of Doxorubicin","volume":"15","author":"Speth","year":"1988","journal-title":"Clin. Pharmacokinet."},{"key":"ref_30","first-page":"2467","article-title":"Mechanism of specific nuclear transport of adriamycin: The mode of nuclear translocation of adriamycin-proteasome complex","volume":"61","author":"Kiyomiya","year":"2001","journal-title":"Cancer Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1038\/s41598-017-01624-4","article-title":"DNA methylation and Transcriptome Changes Associated with Cisplatin Resistance in Ovarian Cancer","volume":"7","author":"Lund","year":"2017","journal-title":"Sci. Rep."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/14\/13\/3068\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:37:52Z","timestamp":1760139472000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/14\/13\/3068"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,22]]},"references-count":31,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["cancers14133068"],"URL":"https:\/\/doi.org\/10.3390\/cancers14133068","relation":{},"ISSN":["2072-6694"],"issn-type":[{"type":"electronic","value":"2072-6694"}],"subject":[],"published":{"date-parts":[[2022,6,22]]}}}