{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T15:17:34Z","timestamp":1791299854644,"version":"4.1.0"},"reference-count":46,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2018,12,5]],"date-time":"2018-12-05T00:00:00Z","timestamp":1543968000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007458","name":"Qatar Foundation","doi-asserted-by":"publisher","award":["BMRP-Bridge Funding Grant"],"award-info":[{"award-number":["BMRP-Bridge Funding Grant"]}],"id":[{"id":"10.13039\/100007458","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Triple negative breast cancers (TNBCs) do not respond to conventional estrogen receptor\/progesterone receptor\/human epidermal growth factor receptor-2 targeted interventions due to the absence of the respective receptor targets. They are aggressive, exhibit early recurrence, metastasize, are more invasive in nature, and develop drug resistance. Some plant-derived substances have been screened and have gained attention as efficient anticancer drugs for TNBCs with few adverse effects. Here, we evaluate triptolide (concentrations in the range of 100 pM to 10 \u00b5M), a di-terpene tri-epoxide isolated from thunder god vine for its efficacy as anticancer drug in MDA-MB-231 TNBC cells. Cell proliferation and viability were assessed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay and trypan blue exclusion assay, respectively. A flow cytometry-based apoptosis assay was performed by using fluorescein isothiocyanate (FITC)-conjugated annexin V and propidium iodide (PI). Western blotting was performed to determine the levels of apoptotic and autophagy proteins such as caspase 3, LC3B and SQSTM1\/p62. Results indicate that in 72 h of 1 nM triptolide treatment, the percentage of cell proliferation in MDA-MB-231 cells declined to 49 \u00b1 18.9% (mean \u00b1 standard deviation (SD)), whereas the proliferation rate did not drop below 80% in MCF-7 cells (non-TNBC cells which express the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2) even at the highest concentration tested (10 \u00b5M). The MDA-MB-468 cells showed a similar trend to MDA-MB-231 cells. Furthermore, triptolide treatment for 72 h significantly decreased cell viability at concentrations above 10 nM. Apoptotic cell death assay in 72 h triptolide-treated MDA-MB-231 cells revealed 29.3 \u00b1 10.57% of early apoptotic cells in comparison to the control group (4.61 \u00b1 2.24%). Cell cycle analysis indicated accumulation of cells in sub G0\/G1 phase, indicating apoptosis. Western blot analysis in the 25 nM triptolide treatment group revealed induction of autophagy as shown by a significant decrease in the levels of autophagy marker p62 (by 0.2-fold p &lt; 0.0001) and with an increase in the levels of LC3B-II (by 8-fold p &lt; 0.05). An increase in the levels of the apoptotic marker cleaved caspase 3 (by 4-fold p &lt; 0.05) was also observed in triptolide treated MDA-MB-231 cells. Our data shows that triptolide could be an efficient anticancer agent in the treatment of TNBCs.<\/jats:p>","DOI":"10.3390\/biom8040163","type":"journal-article","created":{"date-parts":[[2018,12,5]],"date-time":"2018-12-05T12:22:00Z","timestamp":1544012520000},"page":"163","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Triptolide Decreases Cell Proliferation and Induces Cell Death in Triple Negative MDA-MB-231 Breast Cancer Cells"],"prefix":"10.3390","volume":"8","author":[{"given":"Elizabeth","family":"Varghese","sequence":"first","affiliation":[{"name":"Department of Physiology and Biophysics, Weill Cornell Medicine-Qatar, Education City, Qatar Foundation, P. O. Box 24144, Doha, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5541-6623","authenticated-orcid":false,"given":"Samson Mathews","family":"Samuel","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Weill Cornell Medicine-Qatar, Education City, Qatar Foundation, P. O. Box 24144, Doha, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1025-7032","authenticated-orcid":false,"given":"Sharon","family":"Varghese","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Weill Cornell Medicine-Qatar, Education City, Qatar Foundation, P. O. Box 24144, Doha, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5874-4059","authenticated-orcid":false,"given":"Sohaila","family":"Cheema","sequence":"additional","affiliation":[{"name":"Institute for Population Health, Weill Cornell Medicine-Qatar, Education City, Qatar Foundation, P. O. Box 24144, Doha, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ravinder","family":"Mamtani","sequence":"additional","affiliation":[{"name":"Institute for Population Health, Weill Cornell Medicine-Qatar, Education City, Qatar Foundation, P. O. Box 24144, Doha, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5196-3366","authenticated-orcid":false,"given":"Dietrich","family":"B\u00fcsselberg","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Weill Cornell Medicine-Qatar, Education City, Qatar Foundation, P. O. Box 24144, Doha, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,12,5]]},"reference":[{"key":"ref_1","unstructured":"WHO (2018, October 10). Breast Cancer. Available online: http:\/\/www.who.int\/cancer\/prevention\/diagnosis-screening\/breast-cancer\/en\/."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1016\/j.biopha.2018.02.108","article-title":"Chemotherapeutic agents for the treatment of metastatic breast cancer: An update","volume":"101","author":"Abotaleb","year":"2018","journal-title":"Biomed. Pharmacother."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"21","DOI":"10.4103\/0975-7406.92726","article-title":"Application of microarray in breast cancer: An overview","volume":"4","author":"Kumar","year":"2012","journal-title":"J. Pharm. Bioallied Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1038\/415530a","article-title":"Gene expression profiling predicts clinical outcome of breast cancer","volume":"415","author":"Dai","year":"2002","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Varghese, E., Samuel, S.M., Abotaleb, M., Cheema, S., Mamtani, R., and Busselberg, D. (2018). The \u201cyin and yang\u201d of natural compounds in anticancer therapy of triple-negative breast cancers. Cancers, 10.","DOI":"10.3390\/cancers10100346"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1038\/nrclinonc.2016.66","article-title":"Triple-negative breast cancer: Challenges and opportunities of a heterogeneous disease","volume":"13","author":"Bianchini","year":"2016","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_7","first-page":"93","article-title":"Triple-negative breast cancer: Treatment challenges and solutions","volume":"8","author":"Collignon","year":"2016","journal-title":"Breast Cancer (Dove Med. Press)"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3131","DOI":"10.7150\/jca.18457","article-title":"Breast cancer cell line classification and its relevance with breast tumor subtyping","volume":"8","author":"Dai","year":"2017","journal-title":"J. Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1634\/theoncologist.2011-S1-01","article-title":"Triple-negative breast cancer: An unmet medical need","volume":"16","author":"Hudis","year":"2011","journal-title":"Oncologist"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1097\/PPO.0b013e3181d24ff7","article-title":"Triple-negative breast cancer: Role of specific chemotherapy agents","volume":"16","author":"Isakoff","year":"2010","journal-title":"Cancer J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1200\/JOP.2017.022632","article-title":"Practical approach to triple-negative breast cancer","volume":"13","author":"Gadi","year":"2017","journal-title":"J. Oncol. Pract."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.2217\/pgs-2017-0117","article-title":"Triple-negative breast cancer and the potential for targeted therapy","volume":"18","author":"Jhan","year":"2017","journal-title":"Pharmacogenomics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2750","DOI":"10.1172\/JCI45014","article-title":"Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies","volume":"121","author":"Lehmann","year":"2011","journal-title":"J. Clin. Investig."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1002\/path.4280","article-title":"Identification and use of biomarkers in treatment strategies for triple-negative breast cancer subtypes","volume":"232","author":"Lehmann","year":"2014","journal-title":"J. Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Singh, S., Sharma, B., Kanwar, S.S., and Kumar, A. (2016). Lead phytochemicals for anticancer drug development. Front. Plant. Sci., 7.","DOI":"10.3389\/fpls.2016.01667"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1007\/s00394-016-1181-5","article-title":"Oregano demonstrates distinct tumour-suppressive effects in the breast carcinoma model","volume":"56","author":"Kubatka","year":"2017","journal-title":"Eur J. Nutr."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2837","DOI":"10.1111\/jcmm.13197","article-title":"Antineoplastic effects of clove buds (Syzygium aromaticum L.) in the model of breast carcinoma","volume":"21","author":"Kubatka","year":"2017","journal-title":"J. Cell. Mol. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3670","DOI":"10.1016\/j.bbagen.2013.02.008","article-title":"Natural products: A continuing source of novel drug leads","volume":"1830","author":"Cragg","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"838","DOI":"10.5530\/pj.2017.6.131","article-title":"Triptolide mediated amelioration of breast cancer via modulation of molecular pathways","volume":"9","author":"Sarkar","year":"2017","journal-title":"Pharmacogn. J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1111\/j.1365-2125.2012.04221.x","article-title":"Triptolide in the treatment of psoriasis and other immune-mediated inflammatory diseases","volume":"74","author":"Han","year":"2012","journal-title":"Br. J. Clin. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.jep.2014.06.006","article-title":"Triptolide: Progress on research in pharmacodynamics and toxicology","volume":"155","author":"Li","year":"2014","journal-title":"J. Ethnopharmacol."},{"key":"ref_22","first-page":"622","article-title":"Targets and molecular mechanisms of triptolide in cancer therapy","volume":"26","author":"Meng","year":"2014","journal-title":"Chin. J. Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1817","DOI":"10.1016\/j.athoracsur.2015.05.074","article-title":"Triptolide inhibits lung cancer cell migration, invasion, and metastasis","volume":"100","author":"Reno","year":"2015","journal-title":"Ann. Thorac. Surg."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Huang, W., He, T., Chai, C., Yang, Y., Zheng, Y., Zhou, P., Qiao, X., Zhang, B., Liu, Z., and Wang, J. (2012). Triptolide inhibits the proliferation of prostate cancer cells and down-regulates sumo-specific protease 1 expression. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0037693"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.canlet.2012.01.006","article-title":"Synergistic anticancer activity of triptolide combined with cisplatin enhances apoptosis in gastric cancer in vitro and in vivo","volume":"319","author":"Li","year":"2012","journal-title":"Cancer Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"490","DOI":"10.3389\/fphar.2017.00490","article-title":"Effect of the natural product triptolide on pancreatic cancer: A systematic review of preclinical studies","volume":"8","author":"Zhang","year":"2017","journal-title":"Front. Pharmacol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2093","DOI":"10.7150\/jca.16178","article-title":"Anti-cancer and sensibilisation effect of triptolide on human epithelial ovarian cancer","volume":"7","author":"Hu","year":"2016","journal-title":"J. Cancer"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ding, X., Zhang, B., Pei, Q., Pan, J., Huang, S., Yang, Y., Zhu, Z., Lv, Y., and Zou, X. (2014). Triptolide induces apoptotic cell death of human cholangiocarcinoma cells through inhibition of myeloid cell leukemia-1. BMC Cancer, 14.","DOI":"10.1186\/1471-2407-14-271"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2181","DOI":"10.3892\/or.2014.3115","article-title":"Triptolide-induced in vitro and in vivo cytotoxicity in human breast cancer stem cells and primary breast cancer cells","volume":"31","author":"Li","year":"2014","journal-title":"Oncol. Rep."},{"key":"ref_30","first-page":"3413","article-title":"Triptolide induces autophagy and apoptosis through ERK activation in human breast cancer MCF-7 cells","volume":"15","author":"Gao","year":"2018","journal-title":"Exp. Ther. Med."},{"key":"ref_31","first-page":"830","article-title":"Triptolide inhibits vascular endothelial growth factor-mediated angiogenesis in human breast cancer cells","volume":"16","author":"Liu","year":"2018","journal-title":"Exp. Ther. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"23933","DOI":"10.18632\/oncotarget.8207","article-title":"Triptolide has anticancer and chemosensitization effects by down-regulating Akt activation through the MDM2\/REST pathway in human breast cancer","volume":"7","author":"Xiong","year":"2016","journal-title":"Oncotarget"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.bcp.2017.03.001","article-title":"Metformin represses glucose starvation induced autophagic response in microvascular endothelial cells and promotes cell death","volume":"132","author":"Samuel","year":"2017","journal-title":"Biochem. Pharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"26927","DOI":"10.18632\/oncotarget.15879","article-title":"Triptolide suppresses the in vitro and in vivo growth of lung cancer cells by targeting hyaluronan-CD44\/RHAMM signaling","volume":"8","author":"Song","year":"2017","journal-title":"Oncotarget"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"633","DOI":"10.3858\/emm.2012.44.11.072","article-title":"Triptolide inhibits ovarian cancer cell invasion by repression of matrix metalloproteinase 7 and 19 and upregulation of E-cadherin","volume":"44","author":"Zhao","year":"2012","journal-title":"Exp. Mol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2054","DOI":"10.4161\/cbt.8.21.9803","article-title":"Triptolide: An inhibitor of a disintegrin and metalloproteinase 10 (ADAM10) in cancer cells","volume":"8","author":"Soundararajan","year":"2009","journal-title":"Cancer Biol. Ther."},{"key":"ref_37","first-page":"4849","article-title":"Triptolide, a potent anti-cancer compound for the treatment of metastatic breast and ovarian cancer cells","volume":"67","author":"Tan","year":"2007","journal-title":"Cancer Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1007\/s10616-007-9057-4","article-title":"Influence of medium type and serum on MTT reduction by flavonoids in the absence of cells","volume":"52","author":"Talorete","year":"2006","journal-title":"Cytotechnology"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Wang, P., Henning, S.M., and Heber, D. (2010). Limitations of MTT and MTS-based assays for measurement of antiproliferative activity of green tea polyphenols. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0010202"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1016\/j.phymed.2012.02.014","article-title":"Triptolide inhibits colon-rectal cancer cells proliferation by induction of G1 phase arrest through upregulation of p21","volume":"19","author":"Liu","year":"2012","journal-title":"Phytomedicine"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"791","DOI":"10.3892\/mmr.2014.2814","article-title":"Triptolide inhibits cell proliferation and tumorigenicity of human neuroblastoma cells","volume":"11","author":"Yan","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1007\/s11060-012-0885-5","article-title":"Triptolide inhibits proliferation and invasion of malignant glioma cells","volume":"109","author":"Zhang","year":"2012","journal-title":"J. Neurooncol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.bcp.2018.03.030","article-title":"Activation of autophagy by stress-activated signals as a cellular self-defense mechanism against the cytotoxic effects of MBIC in human breast cancer cells in vitro","volume":"152","author":"Hasanpourghadi","year":"2018","journal-title":"Biochem. Pharmacol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1944","DOI":"10.1038\/s41467-018-04070-6","article-title":"Autophagy promotes the survival of dormant breast cancer cells and metastatic tumour recurrence","volume":"9","author":"Vodnala","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1016\/j.jep.2015.01.004","article-title":"Investigation of the active components in Tripterygium wilfordii leading to its acute hepatotoxicty and nephrotoxicity","volume":"162","author":"Li","year":"2015","journal-title":"J. Ethnopharmacol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Li, J., Shen, F., Guan, C., Wang, W., Sun, X., Fu, X., Huang, M., Jin, J., and Huang, Z. (2014). Activation of Nrf2 protects against triptolide-induced hepatotoxicity. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100685"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/8\/4\/163\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:31:24Z","timestamp":1760196684000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/8\/4\/163"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,12,5]]},"references-count":46,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["biom8040163"],"URL":"https:\/\/doi.org\/10.3390\/biom8040163","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,12,5]]}}}