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This study evaluates how myristicin, an aromatic anticancer compound mainly found in nutmeg, induces mitochondrial-mediated apoptosis in MDA-MB-231 TNBC cells. MTT assays quantify anti-proliferative effects, while annexin V/PI flow cytometry measures apoptosis. Protein expression analysis shows reduced Bcl2 and HSP60 alongside increased caspase and apoptosis-related markers, supporting a Bcl2/Bid-linked mechanism. In BALB/c mice, myristicin reduces tumor weight and volume and enhances apoptotic gene, protein, and enzyme profiles, indicating improved in vivo efficacy.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/mitochondrial-mediated-apoptosis-evaluation-of-myristicin-on-triple-negative-breast-cancer-model-via-bcl2bid-pathway/345969/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/mitochondrial-mediated-apoptosis-evaluation-of-myristicin-on-triple-negative-breast-cancer-model-via-bcl2bid-pathway/345969.png","ImageObject",300,407,{"name":92,"@type":93},"Evangeline","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-09-26","2026-09-22",true,{"@type":102,"interactionType":103,"userInteractionCount":81},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"Why is TNBC particularly difficult to treat?","Question",{"text":112,"@type":113},"TNBC lacks estrogen, progesterone, and HER-2 receptors, so targeted therapies are limited and chemotherapy remains the main option. It also shows poor prognosis, higher recurrence, and recurrent metastases.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"What experimental methods were used to assess myristicin’s anticancer effects?",{"text":117,"@type":113},"MTT assays evaluated anti-proliferative potential, while annexin V/PI flow cytometry quantified apoptosis. Protein expression analysis was used to evaluate apoptosis- and mitochondrial-related markers.",{"name":119,"@type":110,"acceptedAnswer":120},"What did the in vivo mouse results show after myristicin treatment?",{"text":121,"@type":113},"In BALB/c mice with 4T1-induced tumors, myristicin treatment reduced tumor weight and volume versus the control group. Enzyme analysis and gene/protein expression indicated significant apoptotic properties.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},345969,1790217833,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":81,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},13056703019662,"https://ap-avatar.wpscdn.com/avatar/be000253a8e92610077?_k=1778726343310543188","Cell Biochemistry and Function   \nRESEARCH ARTICLE   \nMitochondrial Mediated Apoptosis Evaluation of Myristicin on Triple Negative Breast Cancer Model Via Bcl2/Bid Pathway  \nSudhina Sufina Nazar1,2 | Aswathy Vadakkumkattle Ajaykumar3 | Janeesh Plakkal Ayyappan1,2   \n1Translational Nanomedicine and Lifestyle Disease Research Laboratory, Department of Biochemistry, University of Kerala, Kariavattom campus, Thiruvananthapuram, Kerala, India | 2Centre for Advanced Cancer Research, Department of Biochemistry, University of Kerala, Kariavattom campus,  \nThiruvananthapuram, Kerala, India | 3Department of Anatomy, P K Das Institute of Medical Sciences, Vaniyamkulam, Ottapalam, Palakkad, Kerala, India Correspondence: Janeesh Plakkal Ayyappan ([janeeshbiochemistry@keralauniversity.ac.in](janeeshbiochemistry@keralauniversity.ac.in))  \nReceived: 15 February 2026 | Revised: 23 July 2026 | Accepted: 3 August 2026  \nFunding: Indian Council of Medical Research, Grant/Award Numbers: 10.13039/501100001411, 2021‐8110; Start‐up Research Grant from Science and Engineering Research Board (SERB) India, Grant/Award Number: SRG/2021/001831; Start‐up grant for new faculty from University Grants Commission (UGC) India, Grant/Award Number: F.30‐596/2021; University of Kerala research support in the form of Plan and Nonplan funds, Grant/Award Number:  \n7615/2021/UOK; Kerala Biotechnology Commission, Grant/Award Numbers: 10.13039/501100020626, KSCSTE/1025/2023‐YIPBKBC Keywords: apoptosis | mitochondrial stress | myristicin | ROS | TNBC  \nABSTRACT  \nAmong 15‐20 cases of breast cancer, triple‐negative breast cancer (TNBC) is the deadliest form of the disease. The most effective form of treatment for this type of cancer is still targeted chemotherapy because it lacks hormone receptors. Myristicin, an active aromatic compound with anticancer properties, is mostly found in nutmeg. Antitumor, antioxidant, and antimicrobial activity are among few of the numerous properties of myristicin. On TNBC cells, the exact modes of action are mostly unidentified. This study shows that myristicin triggered the mitochondria‐mediated apoptosis in MDA‐MB‐231 cells. The MTT assay assessed the anti‐proliferative potential of myristicin on TNBC cells (IC50 0.65 mM ± 0.98) . Flow cytometry analysis was used to evaluate themyristicin's effects on cell apoptosis using annexin V/PI (46.4 ± 2.31%) . After evaluating the protein expression, myristicinsignificantly decreased the expression of Bcl2 and HSP60 while enhancing the expression of proteins such as caspase 9, caspase 3, bid, bad, caspase 7, P53, cytochrome c, and SDHA. Furthermore, our research confirmed that myristicin has a lower toxicological profile and greater in vivo therapeutic efficacy. In BALB/c mice, 4T1 cells were injected subcutaneously to develop breast tumors, and the mice subsequently received myristicin. According to in vivo results, myristicin treatment reduced tumor weight and volume when compared to the breast cancer control group. Besides, the enzyme analysis, gene, and protein expression showed significant apoptotic properties on myristicin treated group. According to these results, myristicin may be used as a therapeutic approach for the management of TNBC.  \n\n| Abbreviations: ALP, Alkaline Phosphatase; AO/EtBr, Acridine Orange/Ethidium Bromide; BC, Breast cancer Control; CK, Creatine Kinase; DHE, Dihydroethidium; DMSO, dimethyl sulfoxide; FBS, Fetal bovine serum; GST, Glutathione Transferase; LDH, Lactate Dehydrogenase; M, Myristicin; MDA, Malonaldehyde; MTT, 3‐(4,5‐ dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide; PC, Positive Control; ROS, Reactive Oxygen Species; RPMI 1640 Medium, Roswell Park Memorial Institute; SGOT, Serum Glutamic Oxaloacetic Transaminase; SOD, Superoxide Dismutase; TM, Tamoxifen. |\n| --- |\n| This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is pr","cbCaiuRPdpZ6lQ43","https://ap.wps.com/l/cbCaiuRPdpZ6lQ43","pdf",6415059,18,"English","# Abstract\n# Summary\n# Introduction","[{\"question\":\"Why is TNBC particularly difficult to treat?\",\"answer\":\"TNBC lacks estrogen, progesterone, and HER-2 receptors, so targeted therapies are limited and chemotherapy remains the main option. It also shows poor prognosis, higher recurrence, and recurrent metastases.\"},{\"question\":\"What experimental methods were used to assess myristicin’s anticancer effects?\",\"answer\":\"MTT assays evaluated anti-proliferative potential, while annexin V/PI flow cytometry quantified apoptosis. Protein expression analysis was used to evaluate apoptosis- and mitochondrial-related markers.\"},{\"question\":\"What did the in vivo mouse results show after myristicin treatment?\",\"answer\":\"In BALB/c mice with 4T1-induced tumors, myristicin treatment reduced tumor weight and volume versus the control group. Enzyme analysis and gene/protein expression indicated significant apoptotic properties.\"}]","Mitochondrial Mediated Apoptosis Evaluation of Myristicin on Triple Negative Breast Cancer Model Via Bcl2/Bid Pathway | PDF",1790059569,45]