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Hutch (Buxaceae) Exhibits Its Anticancer Potential by Stimulating ROS Production and Caspase p53-BCL-2-Dependent Apoptosis in Hepatocellular Carcinoma and Prostate Cancer Cell Lines","","Buxus natalensis is presented as a triterpenoidal-alkaloid source with reported activity against multiple cancer types, yet its extract’s anticancer effects were previously undocumented. The study evaluated antiproliferative activity of various leaf extracts on eight cancer cell lines, comparing effects with Chang liver cells as a normal control. The hydroethanolic leaf extract (BNHLE) showed strong cytotoxicity, especially in LNCaP and HepG2 cells, and induced apoptosis through ROS generation, caspase activation, p53 upregulation, and NF-κB-p65/BCL-2 suppression in a dose-dependent manner. 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Chang liver cells derived from normal liver tissue served as the control to compare cytotoxicity and selectivity.","Answer",{"name":116,"@type":111,"acceptedAnswer":117},"Which effects did the hydroethanolic leaf extract (BNHLE) produce in LNCaP and HepG2 cells?",{"text":118,"@type":114},"BNHLE inhibited proliferation and exerted cytotoxic effects, with apoptosis induction supported by ROS production, caspase-3/-7 and caspase-9 activity, p53 upregulation, and suppression of NF-κB-p65 and BCL-2.",{"name":120,"@type":111,"acceptedAnswer":121},"How were the phytochemical constituents and potential active compounds identified?",{"text":122,"@type":114},"Phytochemical analysis quantified flavonoids and phenolics, while LC-MS profiling identified multiple compounds including robinin and rutin. The results were used to support the extract’s potential as a source of bioactive anticancer compounds.","https://schema.org",{"og:url":83,"og:type":125,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":127,"canonical":83},"index,follow",{"doc_id":129,"site_id":62},365539,1790219457,{"code":4,"msg":5,"data":132},{"doc_id":129,"user_id":133,"nickname":92,"user_avatar":134,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":135,"file_id":136,"file_url":137,"file_type":138,"file_size":139,"view_count":105,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":140,"language":141,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":142,"faqs":143,"seo_title":144,"seo_description":67,"update_tm":145,"read_time":146},549768072016,"https://ap-avatar.wpscdn.com/davatar_155a257f0dc6eb9ab79c44ca47cae57d","Article  \nBuxus natalensis (Oliv.) Hutch (Buxaceae) Exhibits Its Anticancer Potential by Stimulating ROS Production and  \nCaspase-p53-BCL-2-Dependent Apoptosis in Hepatocellular Carcinoma and Prostate Cancer Cell Lines  \nEmmanuel Mfotie Njoya *, Gaetan T. Tabakam , Chika I. Chukwuma and Tshepiso J. Makhafola *  \nAcademic Editor: Karthik Gourishetti  \nReceived: 31 March 2025  \nRevised: 20 April 2025  \nAccepted: 26 April 2025  \nPublished: 28 April 2025  \nCitation: Mfotie Njoya, E.; Tabakam, G.T.; Chukwuma, C.I.; Makhafola, T.J. Buxus natalensis (Oliv.) Hutch (Buxaceae) Exhibits Its Anticancer Potential by Stimulating ROS Production and Caspase  \np53-BCL-2-Dependent Apoptosis in Hepatocellular Carcinoma and Prostate Cancer Cell Lines. Int. J. Mol. Sci. 2025, 26, 4173. [https://doi.org/](https://doi.org/)[ ](https://doi.org/)[10.3390/ijms26094173](10.3390/ijms26094173)  \nCopyright: © 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://creativecommons.org/](https://creativecommons.org/)[ ](https://creativecommons.org/)[licenses/by/4.0/](licenses/by/4.0/)) .  \nCentre for Quality of Health and Living, Faculty of Health and Environmental Sciences, Central University of  \nTechnology, Bloemfontein 9301, Free State, South Africa; [tgaetan@cut.ac.za](tgaetan@cut.ac.za) (G.T.T.); [cchukwuma@cut.ac.za](cchukwuma@cut.ac.za) (C.I.C.)  \n* Correspondence: [enjoya@cut.ac.za](enjoya@cut.ac.za) (E.M.N.); [jmakhafola@cut.ac.za](jmakhafola@cut.ac.za) (T.J.M.)  \nAbstract: Buxus natalensis is recognized as a rich source of triterpenoidal alkaloids that are known to be effective in fighting different cancer types. Nevertheless, to date, no anticancer potential of B. natalensis extract has been yet described. Here, we investigated the antiproliferative activity of different B. natalensis leaf extracts on eight cancer cell lines (MCF-7, 4T1, Caco-2, HeLa, A549, HepG2, DU145, and LNCaP) . Chang liver cell line derived from normal liver tissue, was used as control. B. natalensis hydroethanolic leaf extract (BNHLE) was found to exert significant cytotoxic effect against cancerous cell lines, with the highest efficacy being observed on LNCaP and HepG2 with IC 50 values of 47.39 and 78.01 µg/mL, respectively. Interestingly, BNHLE was less cytotoxic towards Chang liver cells with an IC50 value of 334.10 µg/mL, yielding selectivity index (SI) values of 6.96 and 4.22 against LNCaP and HepG2 cells, respectively. The study of mechanism of action revealed that BNHLE exerted its antiproliferative effect by inducing ROS production and caspase-3/-7, and-9 activities in LNCaP and HepG2 cells. Moreover, it was found that BNHLE activated apoptosis in both cancerous cell lines by enhancing the expression levels of p53, while suppressing the expression of NF-κB-p65 and BCL-2 protein levels in a dose-dependent manner. The phytochemical analysis of BNHLE showed the presence of flavonoids (24.45 mgQE/g extract) and phenolics (84.64 mgGAE/g extract), and its LC-MS profiling identified several compounds including robinin and rutin, which are known for their cytotoxic effect against different cancer cell lines, such as hepatocellular carcinoma and prostate cancer cell lines. Several compounds are still unknown from B. natalensis, but the data obtained so far justify the use of B. natalensis as a potential source of bioactive compounds against hepatocellular and prostate cancers.  \nKeywords: apoptosis; BAX/BCL-2 ratio; ROS; caspases; Buxus natalensis; phytochemicals; cancer  \n1. Introduction  \nCancer is a group of diseases caused by various risk factors that induce genetic and epigenetic changes leading to abnormal and uncontrolled cell division that can invade adjoining parts of the body and spread to other organs [1,2] . It is one of the leading causes of morbidity and mortality worldwide, with approximately 20 million new cases and","cbCaivUTFsF7zTeF","https://ap.wps.com/l/cbCaivUTFsF7zTeF","pdf",11828096,21,"English","# Introduction\n## Anticancer background and oxidative stress\n## Reactive oxygen species (ROS) in tumor biology\n## Study rationale and therapeutic targeting","[{\"question\":\"What experimental model and control were used to assess Buxus natalensis leaf extracts?\",\"answer\":\"Eight cancer cell lines were treated with different B. natalensis leaf extracts. Chang liver cells derived from normal liver tissue served as the control to compare cytotoxicity and selectivity.\"},{\"question\":\"Which effects did the hydroethanolic leaf extract (BNHLE) produce in LNCaP and HepG2 cells?\",\"answer\":\"BNHLE inhibited proliferation and exerted cytotoxic effects, with apoptosis induction supported by ROS production, caspase-3/-7 and caspase-9 activity, p53 upregulation, and suppression of NF-κB-p65 and BCL-2.\"},{\"question\":\"How were the phytochemical constituents and potential active compounds identified?\",\"answer\":\"Phytochemical analysis quantified flavonoids and phenolics, while LC-MS profiling identified multiple compounds including robinin and rutin. The results were used to support the extract’s potential as a source of bioactive anticancer compounds.\"}]","Buxus natalensis (Oliv.) Hutch (Buxaceae) Exhibits Its Anticancer Potential by Stimulating ROS Production and Caspase p53-BCL-2-Dependent Apoptosis in Hepatocellular Carcinoma and Prostate Cancer Cell Lines | PDF",1790161226,53]