{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T12:44:44Z","timestamp":1784033084482,"version":"3.55.0"},"reference-count":57,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,12,13]],"date-time":"2024-12-13T00:00:00Z","timestamp":1734048000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82472721 82273107 32470571"],"award-info":[{"award-number":["82472721 82273107 32470571"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>\u03b2-caryophyllene (BCP) is a naturally occurring bicyclic sesquiterpene extracted from various plants, and widely used as a medicinal agent for various diseases. During hepatocellular carcinoma (HCC) development, cancer cells generally exhibit increased cell proliferation due to mutations or aberrant expression of key regulatory genes. The current study determines the cytotoxic effects of BCP alone or in combination with doxorubicin (DOX) and cisplatin (DDP) on HCC cells, and elucidates the underlying mechanism of BCP to exert its anticancer activities.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>HepG2, SMMC-7721 HCC cells, and HL-7702 normal liver cells were treated with BCP, DOX, and DDP individually or combinatorially. Cell proliferation assay, flow cytometric assay, and Western blot were employed to evaluate the cytotoxic effects of these treatments. Transwell assays were used to examine BCP\u2019s effects on HCC cell migration and invasion. RNA-seq analysis was used to determine BCP\u2019s primary target genes in HepG2 cells. Integrative analysis of differentially expressed genes (DEGs) of RNA-seq data with an HCC TCGA dataset identified BCP-targeted genes that were verified by RT-qPCR analysis. Ectopic gene expression, cell viability, and colony formation assay were performed to validate the primary targets of BCP.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>BCP selectively inhibited HCC cell proliferation while exhibited relatively low toxicity in normal liver cells; however, DOX and DDP showed higher toxicity in normal cells than that in HCC cells. In combinatorial treatments, BCP synergistically enhanced cytotoxicity of DOX and DDP in HCC cells but this effect was markedly reduced in HL-7702 cells. BCP treatment reduced migration and invasion of HCC cells. Furthermore, RNA-seq analyses of BCP-treated HepG2 cells identified 433 protein-coding DEGs. Integrative analyses revealed five BCP-targeted DEGs regulating the MAPK signaling pathway. Among these five genes, three displayed a significantly positive correlation of their expression with the overall survival of HCC patients. As a primary target, PGF was significantly downregulated by BCP treatment, and its exogenous expression desensitized HCC cells to BCP-mediated inhibition.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>BCP inhibits malignant properties of HCC and synergistically sensitizes the anticancer activity of DOX and DDP. In HCC cells, BCP primarily targets the PGF gene and MAPK signaling pathway.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fphar.2024.1492670","type":"journal-article","created":{"date-parts":[[2024,12,13]],"date-time":"2024-12-13T06:23:57Z","timestamp":1734071037000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":11,"title":["\u03b2-caryophyllene sensitizes hepatocellular carcinoma cells to chemotherapeutics and inhibits cell malignancy through targeting MAPK signaling pathway"],"prefix":"10.3389","volume":"15","author":[{"given":"Irum","family":"Basheer","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hai","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangyue","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shah","family":"Jehan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali","family":"Raza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chentao","family":"Du","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Najeeb","family":"Ullah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dangdang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangchao","family":"Sui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,12,13]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"8354","DOI":"10.3390\/molecules27238354","article-title":"Beta-caryophyllene enhances the anti-tumor activity of cisplatin in lung cancer cell lines through regulating cell cycle and apoptosis signaling molecules","volume":"27","author":"Ahmed","year":"2022","journal-title":"Molecules"},{"key":"B2","doi-asserted-by":"publisher","first-page":"872394","DOI":"10.1155\/2012\/872394","article-title":"\u03b2-Caryophyllene, a compound isolated from the biblical balm of gilead (Commiphora gileadensis), is a selective apoptosis inducer for tumor cell lines","volume":"2012","author":"Amiel","year":"2012","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1530","DOI":"10.2174\/1871520620666200227093216","article-title":"Beta-caryophyllene suppresses ovarian cancer proliferation by inducing cell cycle arrest and apoptosis","volume":"20","author":"Arul","year":"2020","journal-title":"Anti-Cancer Agents Med. Chem."},{"key":"B4","doi-asserted-by":"publisher","first-page":"976","DOI":"10.1158\/1078-0432.CCR-10-2687","article-title":"Placental growth factor upregulation is a host response to antiangiogenic therapy","volume":"17","author":"Bagley","year":"2011","journal-title":"Clin. Cancer Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"66","DOI":"10.4415\/ANN_10_01_09","article-title":"Molecular aspects of tumor cell migration and invasion","volume":"46","author":"Bozzuto","year":"2010","journal-title":"Ann. Ist. Super. Sanita"},{"key":"B6","doi-asserted-by":"publisher","first-page":"566067","DOI":"10.3389\/fneur.2020.566067","article-title":"PKA-dependent membrane surface recruitment of CI-AMPARs is crucial for BCP-mediated protection against post-acute ischemic stroke cognitive impairment","volume":"11","author":"Chen","year":"2020","journal-title":"Front. Neurol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"259","DOI":"10.1007\/bf00685668","article-title":"Quantitation of the synergistic interaction of edatrexate and cisplatin in vitro","volume":"31","author":"Chou","year":"1993","journal-title":"Cancer Chemother. Pharmacol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"413","DOI":"10.3892\/or.2012.2178","article-title":"Increased expression of placental growth factor in high-grade endometrial carcinoma","volume":"29","author":"Coenegrachts","year":"2013","journal-title":"Oncol. Rep."},{"key":"B9","doi-asserted-by":"publisher","first-page":"57","DOI":"10.2217\/hep.15.41","article-title":"Mechanisms of doxorubicin resistance in hepatocellular carcinoma","volume":"3","author":"Cox","year":"2016","journal-title":"Hepat. Oncol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"10550","DOI":"10.3390\/ijms221910550","article-title":"\u03b2-Caryophyllene induces apoptosis and inhibits angiogenesis in colorectal cancer models","volume":"22","author":"Dahham","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1532","DOI":"10.3390\/ijms23031532","article-title":"Pharmacological effects of cisplatin combination with natural products in cancer chemotherapy","volume":"23","author":"Dasari","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1339","DOI":"10.1517\/14728222.2014.948420","article-title":"Placental growth factor in cancer","volume":"18","author":"Dewerchin","year":"2014","journal-title":"Expert Opin. Ther. Targets"},{"key":"B13","doi-asserted-by":"publisher","first-page":"858","DOI":"10.3390\/cells9040858","article-title":"Modulation of STAT3 signaling, cell redox defenses and cell cycle checkpoints by \u03b2-caryophyllene in cholangiocarcinoma cells: possible mechanisms accounting for doxorubicin chemosensitization and chemoprevention","volume":"9","author":"Di Sotto","year":"2020","journal-title":"Cells"},{"key":"B14","doi-asserted-by":"publisher","first-page":"222","DOI":"10.3390\/cells12020222","article-title":"Polydatin inhibits hepatocellular carcinoma cell proliferation and sensitizes doxorubicin and cisplatin through targeting cell mitotic machinery","volume":"12","author":"Farooq","year":"2023","journal-title":"Cells"},{"key":"B15","doi-asserted-by":"publisher","first-page":"3007","DOI":"10.1002\/cam4.816","article-title":"\u03b2-caryophyllene and \u03b2-caryophyllene oxide-natural compounds of anticancer and analgesic properties","volume":"5","author":"Fidyt","year":"2016","journal-title":"Cancer Med."},{"key":"B16","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1016\/j.cell.2007.08.038","article-title":"Anti-PlGF inhibits growth of VEGF (R)-inhibitor-resistant tumors without affecting healthy vessels","volume":"131","author":"Fischer","year":"2007","journal-title":"Cell"},{"key":"B17","doi-asserted-by":"publisher","first-page":"5420","DOI":"10.3390\/app9245420","article-title":"\u03b2-Caryophyllene: a sesquiterpene with countless biological properties","volume":"9","author":"Francomano","year":"2019","journal-title":"Appl. Sci."},{"key":"B18","doi-asserted-by":"publisher","first-page":"102925","DOI":"10.1016\/j.bioorg.2019.102925","article-title":"Cisplatin: the first metal based anticancer drug","volume":"88","author":"Ghosh","year":"2019","journal-title":"Bioorg. Chem."},{"key":"B19","doi-asserted-by":"publisher","first-page":"58","DOI":"10.4103\/pm.pm_550_19","article-title":"\u03b2-caryophyllene, a natural bicyclic sesquiterpene, induces apoptosis by inhibiting inflammation-associated proliferation in MOLT-4 leukemia cells","volume":"17","author":"Gu","year":"2021","journal-title":"Pharmacogn. Mag."},{"key":"B20","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/j.retram.2017.11.002","article-title":"High MYBL2 expression and transcription regulatory activity is associated with poor overall survival in patients with hepatocellular carcinoma","volume":"66","author":"Guan","year":"2018","journal-title":"Curr. Res. Transl. Med."},{"key":"B21","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1016\/B978-0-12-802147-7.00053-X","article-title":"Cannabis sativa and hemp","volume-title":"Nutraceuticals","author":"Hartsel","year":"2016"},{"key":"B22","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/j.jhep.2012.09.032","article-title":"The placental growth factor as a target against hepatocellular carcinoma in a diethylnitrosamine-induced mouse model","volume":"58","author":"Heindryckx","year":"2013","journal-title":"J. Hepatol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"139","DOI":"10.32604\/or.2023.030190","article-title":"Cisplatin-induced activation of TGF-\u03b2 signaling contributes to drug resistance","volume":"32","author":"Imatsuji","year":"2024","journal-title":"Oncol. Res."},{"key":"B24","doi-asserted-by":"publisher","first-page":"555283","DOI":"10.3389\/fphar.2020.555283","article-title":"Thymoquinone selectively induces hepatocellular carcinoma cell apoptosis in synergism with clinical therapeutics and dependence of p53 status","volume":"11","author":"Jehan","year":"2020","journal-title":"Front. Pharmacol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1038\/nmeth.3317","article-title":"HISAT: a fast spliced aligner with low memory requirements","volume":"12","author":"Kim","year":"2015","journal-title":"Nat. methods"},{"key":"B26","doi-asserted-by":"publisher","first-page":"396","DOI":"10.1016\/j.bbadis.2009.12.009","article-title":"Pathological roles of MAPK signaling pathways in human diseases","volume":"1802","author":"Kim","year":"2010","journal-title":"Biochim. Biophys. Acta"},{"key":"B27","doi-asserted-by":"publisher","first-page":"1295","DOI":"10.1080\/17474124.2021.1991792","article-title":"Current epidemiology in hepatocellular carcinoma","volume":"15","author":"Konyn","year":"2021","journal-title":"Expert Rev. Gastroenterol. Hepatol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1038\/nmeth.1923","article-title":"Fast gapped-read alignment with Bowtie 2","volume":"9","author":"Langmead","year":"2012","journal-title":"Nat. Methods."},{"key":"B29","doi-asserted-by":"publisher","first-page":"323","DOI":"10.1186\/1471-2105-12-323","article-title":"RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome","volume":"12","author":"Li","year":"2011","journal-title":"BMC Bioinform"},{"key":"B30","doi-asserted-by":"publisher","first-page":"16018","DOI":"10.1038\/nrdp.2016.18","article-title":"Hepatocellular carcinoma","volume":"2","author":"Llovet","year":"2016","journal-title":"Nat. Rev. Dis. Prim."},{"key":"B31","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/j.bbcan.2003.09.002","article-title":"Genetic dissection of tumor angiogenesis: are PlGF and VEGFR-1 novel anti-cancer targets?","volume":"1654","author":"Luttun","year":"2004","journal-title":"BBA - Rev. Cancer"},{"key":"B32","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1002\/bimj.201400023","article-title":"On the use of partial area under the ROC curve for comparison of two diagnostic tests","volume":"57","author":"Ma","year":"2015","journal-title":"Biom. J."},{"key":"B33","first-page":"130","article-title":"\u03b2-caryophyllene and low-doses of doxorubicin against liver cancer cells: a \u201cmetronomic chemotherapy\u201d","volume":"122","author":"Mancinelli","year":"2017","journal-title":"Ital. J. Anat. Embryol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"723","DOI":"10.1002\/hep.29913","article-title":"Diagnosis, staging, and management of hepatocellular carcinoma: 2018 practice guidance by the American Association for the Study of Liver Diseases","volume":"68","author":"Marrero","year":"2018","journal-title":"Hepatology"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.31300\/ctbr.20.2019.1-16","article-title":"\u03b2-caryophyllene enhances the transcriptional upregulation of cholesterol biosynthesis in breast cancer cells","volume":"20","author":"Mboge","year":"2019","journal-title":"Curr. Top. Biochem. Res."},{"key":"B36","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1016\/j.cbi.2019.02.028","article-title":"\u03b2-Caryophyllene, a natural bicyclic sesquiterpene attenuates doxorubicin-induced chronic cardiotoxicity via activation of myocardial cannabinoid type-2 (CB2) receptors in rats","volume":"304","author":"Meeran","year":"2019","journal-title":"Chem. Biol. Interact."},{"key":"B37","doi-asserted-by":"publisher","first-page":"3026","DOI":"10.3390\/cancers13123026","article-title":"MAPK\/ERK signaling pathway in hepatocellular carcinoma","volume":"13","author":"Moon","year":"2021","journal-title":"Cancers (Basel)"},{"key":"B38","doi-asserted-by":"publisher","first-page":"6675762","DOI":"10.1155\/2021\/6675762","article-title":"Emerging roles of impaired autophagy in fatty liver disease and hepatocellular carcinoma","volume":"2021","author":"Niture","year":"2021","journal-title":"Int. J. Hepatol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"2819","DOI":"10.1016\/j.ejca.2005.07.022","article-title":"Placenta growth factor is over-expressed and has prognostic value in human breast cancer","volume":"41","author":"Parr","year":"2005","journal-title":"Eur. J. Cancer"},{"key":"B40","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1016\/j.phymed.2018.05.001","article-title":"Synergistic interaction of \u03b2-caryophyllene with aromadendrene oxide 2 and phytol induces apoptosis on skin epidermoid cancer cells","volume":"47","author":"Pavithra","year":"2018","journal-title":"Phytomedicine"},{"key":"B41","doi-asserted-by":"publisher","first-page":"148","DOI":"10.1080\/13813455.2019.1669057","article-title":"\u03b2-Caryophyllene promotes oxidative stress and apoptosis in KB cells through activation of mitochondrial-mediated pathway - an in-vitro and in-silico study","volume":"128","author":"Ramachandhiran","year":"2022","journal-title":"Arch. Physiol. Biochem."},{"key":"B42","doi-asserted-by":"publisher","first-page":"249","DOI":"10.5114\/ceh.2021.109312","article-title":"Overview of lenvatinib as a targeted therapy for advanced hepatocellular carcinoma","volume":"7","author":"Rehman","year":"2021","journal-title":"Clin. Exp. Hepatol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"203","DOI":"10.32074\/1591-951x-295","article-title":"Hepatocellular carcinoma: a clinical and pathological overview","volume":"113","author":"Renne","year":"2021","journal-title":"Pathologica"},{"key":"B44","doi-asserted-by":"publisher","first-page":"427","DOI":"10.1159\/000499765","article-title":"Systemic treatment options in hepatocellular carcinoma","volume":"8","author":"Rimassa","year":"2019","journal-title":"Liver cancer"},{"key":"B45","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1038\/s41590-018-0044-z","article-title":"The immunology of hepatocellular carcinoma","volume":"19","author":"Ringelhan","year":"2018","journal-title":"Nat. Immunol."},{"key":"B46","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1093\/bioinformatics\/btp616","article-title":"edgeR: a Bioconductor package for differential expression analysis of digital gene expression data","volume":"26","author":"Robinson","year":"2010","journal-title":"Bioinformatics"},{"key":"B47","doi-asserted-by":"publisher","first-page":"1089","DOI":"10.1007\/s12325-008-0113-z","article-title":"Systemic therapy of hepatocellular carcinoma: are we making progress?","volume":"25","author":"Roxburgh","year":"2008","journal-title":"Adv. Ther."},{"key":"B48","doi-asserted-by":"publisher","first-page":"292","DOI":"10.1007\/s12012-022-09721-1","article-title":"Doxorubicin-induced cardiotoxicity: an overview on pre-clinical therapeutic approaches","volume":"22","author":"Sheibani","year":"2022","journal-title":"Cardiovasc. Toxicol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"119527","DOI":"10.1016\/j.lfs.2021.119527","article-title":"A comprehensive review on time-tested anticancer drug doxorubicin","volume":"278","author":"Sritharan","year":"2021","journal-title":"Life Sci."},{"key":"B50","first-page":"865","article-title":"Drug synergism: its detection and applications","volume":"298","author":"Tallarida","year":"2001","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"B51","doi-asserted-by":"publisher","first-page":"2120","DOI":"10.1016\/j.ajpath.2012.01.037","article-title":"Yin Yang 1 plays an essential role in breast cancer and negatively regulates p27","volume":"180","author":"Wan","year":"2012","journal-title":"Am. J. Pathol."},{"key":"B52","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1136\/gut.2004.050831","article-title":"Placenta growth factor expression is correlated with survival of patients with colorectal cancer","volume":"54","author":"Wei","year":"2005","journal-title":"Gut"},{"key":"B53","doi-asserted-by":"publisher","first-page":"1912","DOI":"10.3390\/ijms23031912","article-title":"Mitochondrial-targeted therapy for doxorubicin-induced cardiotoxicity","volume":"23","author":"Wu","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B54","doi-asserted-by":"publisher","first-page":"100141","DOI":"10.1016\/j.xinn.2021.100141","article-title":"clusterProfiler 4.0: a universal enrichment tool for interpreting omics data","volume":"2","author":"Wu","year":"2021","journal-title":"innovation"},{"key":"B55","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.archoralbio.2018.01.009","article-title":"Inhibitory effects of \u03b2-caryophyllene on Streptococcus mutans biofilm","volume":"88","author":"Yoo","year":"2018","journal-title":"Arch. Oral Biol."},{"key":"B56","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1016\/bs.acr.2021.03.008","article-title":"Cisplatin chemotherapy and renal function","volume":"152","author":"Zhang","year":"2021","journal-title":"Adv. Cancer Res."},{"key":"B57","doi-asserted-by":"publisher","first-page":"152921","DOI":"10.1016\/j.phymed.2019.152921","article-title":"Ellagic acid synergistically potentiates inhibitory activities of chemotherapeutic agents to human hepatocellular carcinoma","volume":"59","author":"Zhong","year":"2019","journal-title":"Phytomedicine"}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2024.1492670\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,13]],"date-time":"2024-12-13T06:24:02Z","timestamp":1734071042000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2024.1492670\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,13]]},"references-count":57,"alternative-id":["10.3389\/fphar.2024.1492670"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2024.1492670","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,13]]},"article-number":"1492670"}}