{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T15:01:52Z","timestamp":1788966112765,"version":"build-2803163510"},"reference-count":37,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2015,4,10]],"date-time":"2015-04-10T00:00:00Z","timestamp":1428624000000},"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>This study aimed to evaluate the cytotoxicity of a crude extract of Piper cubeba against normal and breast cancer cell lines. To prepare the extract, P. cubeba seeds were ground, soaked in methanol and dichloromethane and isolated by column chromatography. Fractions were tested for cytotoxicity effects on normal fibroblast (L929), normal breast (MCF-12A) and breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231).  The most effective fraction was selected for DNA fragmentation assay to detect apoptotic activity. The results showed that the methanolic crude extract had a higher cytotoxic activity against MDA-MB-468 and MCF-7 than a dichloromethane crude extract. Then, the methanolic crude extract was separated into six fractions, designated A to F. Fraction C was highly active against breast cancer cell lines with an IC50 value less than 4 \u03bcg\/mL. Therefore, Fraction C was further separated into seven fractions, CA to CG. The 1H-NMR profile showed that Fraction CE was long chain hydrocarbons. Moreover, Fraction CE demonstrated the highest activity against MCF-7 cells with an IC50 value of 2.69 \u00b1 0.09 \u03bcg\/mL and lower cytotoxicity against normal fibroblast L929 cells with an IC50 value of 4.17 \u00b1 0.77 \u03bcg\/mL. Finally, DNA fragmentation with a ladder pattern characteristic of apoptosis was observed in MCF-7, MDA-MB-468, MDA-MB-231 and L929 cells, but not in MCF-12A cells.<\/jats:p>","DOI":"10.3390\/nu7042707","type":"journal-article","created":{"date-parts":[[2015,4,10]],"date-time":"2015-04-10T17:06:18Z","timestamp":1428685578000},"page":"2707-2718","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":62,"title":["Cytotoxic Activity of Piper cubeba Extract in Breast Cancer  Cell Lines"],"prefix":"10.3390","volume":"7","author":[{"given":"Potchanapond","family":"Graidist","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand"},{"name":"The Cancer Molecular Biology Excellence Research Laboratory, Prince of Songkla University,  Hat Yai, Songkhla 90110, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mananya","family":"Martla","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaowapa","family":"Sukpondma","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai,  Songkhla 90110, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2015,4,10]]},"reference":[{"key":"ref_1","first-page":"9","article-title":"Cancer Statistics, 2014","volume":"64","author":"Siegel","year":"2014","journal-title":"CA: Cancer J. Clin."},{"key":"ref_2","unstructured":"Cancer Research UK. Available online: http:\/\/cancerhelp.cancerresearchuk.org\/about-cancer\/treatment\/cancer-drugs\/side-effects."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1038\/sj.bjc.6604510","article-title":"Resistance to chemotherapy: New treatments and novel insights into an old problem","volume":"99","author":"Raguz","year":"2008","journal-title":"Br. J. Cancer"},{"key":"ref_4","unstructured":"Sastroamidjojo, S. (1997). ObatAsli Indonesia, Dian Rakyat."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"773","DOI":"10.4103\/1319-2442.98159","article-title":"Nephroprotective effect of Kabab chini (Piper cubeba) in gentamycin-induced nephrotoxicity","volume":"23","author":"Ahmad","year":"2012","journal-title":"Saudi J. Kidney Dis. Transplant."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/S0378-8741(03)00280-0","article-title":"Investigations of anti-inflammatory and anti-nociceptive activities of Piper cubeba, Physalis angulata and Rosa hybrid","volume":"89","author":"Choi","year":"2003","journal-title":"J. Ethnopharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.phcgj.2012.12.001","article-title":"Anti-inflammatory and analgesic evaluation of hydroalcoholic extract and fractions from seeds of Piper cubeba L. (Piperaceae)","volume":"5","author":"Perazzo","year":"2013","journal-title":"Pharmacogn. J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1007\/s11418-007-0159-2","article-title":"Antileishmanial amides and lignans from Piper cubeba and Piper retrofragtum","volume":"61","author":"Bodiwala","year":"2007","journal-title":"J. Nat. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1590\/S1415-47572007000400025","article-title":"Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system","volume":"30","author":"Junqueira","year":"2007","journal-title":"Gen. Mol. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1055\/s-2007-993758","article-title":"Piper cubeba targets multiple aspects of the androgen-signalling pathway. A potential phytotherapy against prostate cancer growth?","volume":"74","author":"Yam","year":"2008","journal-title":"Planta Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1002\/ptr.939","article-title":"Antimycobacterial physalins from Physalis angulata L. (Solanaceae)","volume":"16","author":"Januario","year":"2002","journal-title":"Phytother. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.phymed.2004.06.022","article-title":"CYP3A4 and CYP2D6 inhibitory activities of Indonesian medicinal plants","volume":"13","author":"Usia","year":"2006","journal-title":"Phytomedicine"},{"key":"ref_13","first-page":"153","article-title":"Phytochemical evaluation and antioxidant activity of Piper cubeba and Piper nigrum","volume":"1","author":"Nahak","year":"2011","journal-title":"J. Appl. Pharm. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/j.bse.2007.01.003","article-title":"Lignan profile of Piper cubeba, an Indonesian medicinal plant","volume":"35","author":"Elfahmi","year":"2007","journal-title":"Biochem. Syst. Ecol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/S0378-8741(01)00171-4","article-title":"Anti-inflammatory activity of cubebin, a lignan from the leaves of Zanthoxylum naranjillo Griseb","volume":"75","author":"Bastos","year":"2001","journal-title":"J. Ethnopharmacol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1002\/ptr.2088","article-title":"Evaluation of Piper cubeba extract, (\u2212)-cubebin and its semi-synthetic derivatives against oral pathogens","volume":"21","author":"Silva","year":"2007","journal-title":"Phytother. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1016\/j.fct.2013.01.050","article-title":"A comparative study of the modulatory effects of (\u2212)-cubebin on the mutagenicity\/recombinogenicity induced by different chemical agents","volume":"55","author":"Munari","year":"2013","journal-title":"Food Chem. Toxicol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.jep.2005.01.064","article-title":"Cytotoxic of plants from Malaysia and Thailand used traditionally to treat cancer","volume":"100","author":"Lee","year":"2005","journal-title":"J. Ethnopharmacol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"921","DOI":"10.4314\/tjpr.v13i6.14","article-title":"Cytotoxicity screening of plants of genus Piper in breast cancer cell lines","volume":"13","author":"Sriwiriyajan","year":"2014","journal-title":"Trop. J. Pharm. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"5165","DOI":"10.1021\/jf010697n","article-title":"Antioxidant activity and phenolic compounds in selected herbs","volume":"49","author":"Zheng","year":"2001","journal-title":"J. Agric. Food Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/0891-5849(95)02014-4","article-title":"Anti-oxidant properties of hydroxy-flavones","volume":"20","author":"Cotelle","year":"1996","journal-title":"Free Radic. Biol. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/0027-5107(95)00099-2","article-title":"Natural antimutagenic agents","volume":"350","author":"Mitscher","year":"1996","journal-title":"Mutat. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.1016\/S0959-8049(00)00103-9","article-title":"The antioxidant\/anticancer potential of phenolic compounds isolated from olive oil","volume":"36","author":"Owen","year":"2000","journal-title":"Eur. J. Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1211\/0022357021778600","article-title":"Anti-inflammatory and antioxidant properties of Helichrysum italicum","volume":"54","author":"Sala","year":"2002","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2157","DOI":"10.1016\/j.lfs.2003.09.047","article-title":"Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer","volume":"74","author":"Cai","year":"2004","journal-title":"Life Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1002\/(SICI)1099-1026(199909\/10)14:5<279::AID-FFJ821>3.0.CO;2-0","article-title":"Essential oil constituents of some Piper species","volume":"14","author":"Sumathykutty","year":"1999","journal-title":"Flavour Fragr. J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1819","DOI":"10.3390\/molecules17021819","article-title":"Chemical and biological analyses of the essential oils and main constituents of Piper species","volume":"17","author":"Amaral","year":"2012","journal-title":"Molecules"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1016\/j.bmcl.2004.12.035","article-title":"Synthesis and biological activity evaluation of lignan lactones derived from (\u2212)-cubebin","volume":"15","author":"Pereira","year":"2005","journal-title":"Bioorganic Med. Chem. Lett."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jfca.2012.12.003","article-title":"Effects of (\u2212)-cubebin (Piper cubeba) on cytotoxicity, mutagenicity and expression of p38 MAP kinase and GSTa2 in a hepatoma cell line","volume":"30","author":"Niwa","year":"2013","journal-title":"J. Food Compos. Anal."},{"key":"ref_30","unstructured":"(1992). Biological Evaluation of Medical Devices-Part 5. Tests for Cytotoxicity: In vitro Methods, ISO."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.1177\/00220345950740081301","article-title":"Response of L-929 fibroblasts, human gingival fibroblasts, and human tissue mast cells to various metal cations","volume":"74","author":"Schedle","year":"1995","journal-title":"J. Dent. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1677\/erc.1.01172","article-title":"p53 and breast cancer, an update","volume":"13","author":"Lacroix","year":"2006","journal-title":"Endocr. Relat. Cancer"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.molonc.2007.02.004","article-title":"The morphologies of breast cancer cell lines in three-dimensional assays correlate with their profiles of gene expression","volume":"1","author":"Kenny","year":"2007","journal-title":"Mol. Oncol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1016\/S0008-6363(99)00384-3","article-title":"Morphologic and biochemical hallmarks of apoptosis","volume":"45","author":"Saraste","year":"2000","journal-title":"Cardiovasc. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1080\/01926230701320337","article-title":"Apoptosis: A review of programmed cell death","volume":"35","author":"Elmore","year":"2007","journal-title":"Toxicol. Pathol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.sajb.2012.03.010","article-title":"Aqueous extract of Plinia edulis leaves: Antioxidant activity and cytotoxicity to human breast cancer MCF-7 cell line","volume":"81","author":"Carvalho","year":"2012","journal-title":"S. Afr. J. Bot."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.sajb.2014.03.011","article-title":"Ethanolic extract of Mimosa caesalpiniifolia leaves: Chemical characterization and cytotoxic effect on human breast cancer MCF-7 cell line","volume":"93","author":"Silva","year":"2014","journal-title":"S. Afr. J. Bot."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/7\/4\/2707\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T20:44:34Z","timestamp":1760215474000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/7\/4\/2707"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,4,10]]},"references-count":37,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2015,4]]}},"alternative-id":["nu7042707"],"URL":"https:\/\/doi.org\/10.3390\/nu7042707","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,4,10]]}}}