SavedAdd your journal

Journals / International Journal of Secondary Metabolite / 2026 / Volume: 13 Issue: 2

Apoptosis-related gene modulation by Juglans regia leaf extract in breast and colon cancer cells

Derya Okuyan(BALIKESİR ÜNİVERSİTESİ, SUSURLUK MESLEK YÜKSEKOKULU, VETERİNERLİK BÖLÜMÜ)0000-0001-6758-8556
Pages
—
Publication Date
2026-06-09
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

J. regia may represent a promising natural source for developing novel anticancer agents.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Juglans regia L. (walnut) leaves are rich in phenolic compounds, flavonoids, and terpenoids known for their antioxidant and pharmacological activities.
📊
Key Findings & Data:This study aimed to investigate the cytotoxic and apoptotic effects of ethanol extracts of J. regia leaves on human breast adenocarcinoma (MDA-MB-231), human colorectal adenocarcinoma (HT-29), and human keratinocyte (HaCaT) cell lines.
💡
Conclusion & Impact:J. regia may represent a promising natural source for developing novel anticancer agents.
🔬Design:Lab & Field Experiment
📍Scope:Comprehensive Sample
⚙️Variables:Juglans regia · Apoptosis · Bax

Abstract

Juglans regia L. (walnut) leaves are rich in phenolic compounds, flavonoids, and terpenoids known for their antioxidant and pharmacological activities. Although J. regia has been widely used in traditional medicine, its potential anticancer mechanisms on different human cell lines remain to be fully elucidated. This study aimed to investigate the cytotoxic and apoptotic effects of ethanol extracts of J. regia leaves on human breast adenocarcinoma (MDA-MB-231), human colorectal adenocarcinoma (HT-29), and human keratinocyte (HaCaT) cell lines. Dried J. regia leaves collected from Balıkesir, Türkiye, were extracted with ethanol using the Soxhlet method. Cytotoxicity was evaluated by MTT assay in a dose- and time-dependent manner (1000–50 μg/μL concentrations; 24, 48, and 72 h). Gene expression analysis of apoptosis-related Bax, Bcl-2, and cell cycle regulator p27 was performed by quantitative real-time PCR (qRT-PCR). The ethanol extract of J. regia leaves exhibited a significant antiproliferative effect on MDA-MB-231 and HT-29 cells, while it showed no cytotoxicity on HaCaT cells. The effective concentration was determined as 100 μg/μL. qRT-PCR analysis revealed upregulation of pro-apoptotic Bax and p27 genes and downregulation of anti-apoptotic Bcl-2 in MDA-MB-231 and HT-29 cells in a time-dependent manner, suggesting induction of apoptosis and cell cycle arrest. These findings demonstrate that ethanol extracts of J. regia leaves possess potent anticancer and antiproliferative activities, likely mediated through the modulation of apoptotic pathways and regulation of the mitotic phase. J. regia may represent a promising natural source for developing novel anticancer agents.

Özet

Juglans regia L. (walnut) leaves are rich in phenolic compounds, flavonoids, and terpenoids known for their antioxidant and pharmacological activities. Although J. regia has been widely used in traditional medicine, its potential anticancer mechanisms on different human cell lines remain to be fully elucidated. This study aimed to investigate the cytotoxic and apoptotic effects of ethanol extracts of J. regia leaves on human breast adenocarcinoma (MDA-MB-231), human colorectal adenocarcinoma (HT-29), and human keratinocyte (HaCaT) cell lines. Dried J. regia leaves collected from Balıkesir, Türkiye, were extracted with ethanol using the Soxhlet method. Cytotoxicity was evaluated by MTT assay in a dose- and time-dependent manner (1000–50 μg/μL concentrations; 24, 48, and 72 h). Gene expression analysis of apoptosis-related Bax, Bcl-2, and cell cycle regulator p27 was performed by quantitative real-time PCR (qRT-PCR). The ethanol extract of J. regia leaves exhibited a significant antiproliferative effect on MDA-MB-231 and HT-29 cells, while it showed no cytotoxicity on HaCaT cells. The effective concentration was determined as 100 μg/μL. qRT-PCR analysis revealed upregulation of pro-apoptotic Bax and p27 genes and downregulation of anti-apoptotic Bcl-2 in MDA-MB-231 and HT-29 cells in a time-dependent manner, suggesting induction of apoptosis and cell cycle arrest. These findings demonstrate that ethanol extracts of J. regia leaves possess potent anticancer and antiproliferative activities, likely mediated through the modulation of apoptotic pathways and regulation of the mitotic phase. J. regia may represent a promising natural source for developing novel anticancer agents.

🔖

Cite This Article

Okuyan, D. (2026). Apoptosis-related gene modulation by Juglans regia leaf extract in breast and colon cancer cells. International Journal of Secondary Metabolite, Volume: 13 Issue: 2. https://doi.org/10.21448/ijsm.1821104

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (29)

  1. Alshatwi, A. (2011). Anti-proliferative effects of organic extracts from root bark of Juglans Regia L. (RBJR) on MDA-MB-231 human breast cancer cells: role of Bcl-2/Bax, caspases and Tp53. Asian Pac J Cancer Prev, 12(2), 525-530. https://doi.org/10.7314/APJCP.2011.12.2.525
  2. Babacan, D. (2023). BCL-2 Protein family and cancer. Journal of the Institute of Science of Yüzüncü Yil University, 28(3), 1218-1232. https://doi.org/10.53433/yyufbed.1220984
  3. Carocho, M., & Ferreira, I.C. (2013). A review on antioxidants, prooxidants and related controversy: Natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food and Chemical Toxicology, 51, 15-25. https://doi.org/10.1016/j.fct.2012.09.021
  4. Carvalho, M., Ferreira, P.J., Mendes, V.S., Silva, R., Pereira, J.A., Jerónimo, C., & Silva, B.M. (2010). Human cancer cell antiproliferative and antioxidant activities of Juglans regia L. Food and Chemical Toxicology, 48(1), 441-447. https://doi.org/10.1016/j.fct.2009.10.043
  5. Cosmulescu, S.N., Trandafir, I., Achim, G., & Baciu, A. (2011). Juglone content in leaf and green husk of five walnut (Juglans regia L.) cultivars. Notulae Botanicae Horti Agrobotanici Cluj Napoca, 39(1), 237-240. https://doi.org/1 10.15835/nbha3912378
  6. Cragg, G.M., Boyd, M.R., Khanna, R., Kneller, R., Mays, T.D., Mazan, K.D.,. & Sausville, E.A. (1999). International collaboration in drug discovery and development: the NCI experience. Pure and Applied Chemistry, 71(9), 1619-1633. https://doi.org/10.1351/pac199971091619
  7. Cragg, G.M., & Newman, D.J. (2005). Plants as a source of anti cancer agents. Journal of Ethnopharmacology, 100(1-2), 72-79. https://doi.org/10.1016/j.jep.2005.05.011
  8. Elouafy, Y., El Yadini, A., El Moudden, H., Harhar, H., Alshahrani, M.M., Awadh, A.A.A.,. & Tabyaoui, M. (2022). Influence of the extraction method on the quality and chemical composition of walnut (Juglans regia L.) oil. Molecules, 27(22), 7681. https://doi.org/10.3390/molecules27227681
  9. Jenab, M., Ferrari, P., Slimani, N., Norat, T., Casagrande, C., Overad, K.,. & Riboli, E. (2004). Association of nut and seed intake with colorectal cancer risk in the European Prospective Investigation into Cancer and Nutrition. Cancer Epidemiology Biomarkers & Prevention, 13(10), 1595-1603. https://doi.org/10.1158/1055-9965.EPI-04-0161.
  10. Li, W., Li, D.Y., Wang, H.D., Zheng, Z.J., Hu, J., & Li, Z.Z. (2015). Juglans regia hexane extract exerts antitumor effect, apoptosis induction and cell circle arrest in prostate cancer cells in vitro. Tropical Journal of Pharmaceutical Research, 14(3), 399 405. https://doi.org/10.4314/tjpr.v14i3.7
  11. Oliveira, I., Sousa, A., Ferreira, I.C., Bento, A., Estevinho, L., & Pereira, J.A. (2008). Total phenols, antioxidant potential and antimicrobial activity of walnut (Juglans regia L.) green husks. Food and Chemical Toxicology, 46(7), 2326-2331. https://doi.org/10.1016/j.fct.2008.03.017
  12. Marimuthu, P. (2008). Projection of cancer incidence in five cities and cancer mortality in India. Indian Journal of Cancer, 45(1), 4-7. https://doi.org/10.4103/0019-509X.40639
  13. Mathew, A., Peters, U., Chatterjee, N., Kulldorff, M., & Sinha, R. (2004). Fat, fiber, fruits, vegetables, and risk of colorectal adenomas. International Journal of Cancer, 108(2), 287 292. https://doi.org/10.1002/ijc.10984
  14. Nahrstedt, A., Vetter, U., Hammerschmidt, F.J. (1981). Composition of the steam distillation product from the leaves of juglans regia (author’s transl). Planta Med, 42, 313–332.
  15. Nour, V., Trandafir, I., & Cosmulescu, S. (2013). HPLC determination of phenolic acids, flavonoids and juglone in walnut leaves. Journal of Chromatographic Science, 51(9), 883 890. https://doi.org/10.1093/chromsci/bmt090
  16. Pereira, J.A., Oliveira, I., Sousa, A., Valentão, P., Andrade, P.B., Ferreira, I.C.,. & Estevinho, L. (2007). Walnut (Juglans regia L.) leaves: Phenolic compounds, antibacterial activity and antioxidant potential of different cultivars. Food and Chemical Toxicology, 45(11), 2287 2295. https://doi.org/10.1016/j.fct.2007.06.004
  17. Pires, T.C., Dias, M.I., Barros, L., Alves, M.J., Oliveira, M.B.P., Santos-Buelga, C., & Ferreira, I.C. (2018). Antioxidant and antimicrobial properties of dried Portuguese apple variety (Malus domestica Borkh. Cv Bravo de Esmolfe). Food Chemistry, 240, 701 706. https://doi.org/10.1016/j.foodchem.2017.08.010
  18. Reddy, L.A.L.I.N.I., Odhav, B., & Bhoola, K.D. (2003). Natural products for cancer prevention: a global perspective. Pharmacology & Therapeutics, 99(1), 1 13. https://doi.org/10.1016/S0163 7258(03)00042-1.
  19. Türkoğlu, S.A., Koç, S., Poyrazlı, F., & Selvi, S. (2024). Cytotoxic effect of Trigonella coerulescens subsp. ayvalikensis Erdoğan, Selvi & Tümen in prostate and colon cancer cell lines. International Journal of Secondary Metabolite, 11(4), 700-709. https://doi.org/10.21448/ijsm.1485930
  20. Verhoeckx, K., Cotter, P., López-Expósito, I., Kleiveland, C., Lea, T., Mackie, A.,. & Wichers, H. (2015). The impact of food bioactives on health: in vitro and ex vivo models. https://doi.org/10.1007/978-3-319-16104-4
  21. Verpoorte, R. (1998). Exploration of nature's chemodiversity: the role of secondary metabolites as leads in drug development. Drug Discovery Today, 3(5), 232-238.
  22. Yang, J., Liu, R.H., Halim, L. (2009). Antioxidant and antiproliferative activities of common edible nut seeds. Food Sci Technol, 42, 1–8. https://doi.org/10.1016/j.lwt.2008.07.007.
  23. Żurek, N., Pycia, K., Pawłowska, A., Potocki, L., & Kapusta, I.T. (2023). Chemical profiling, bioactive properties, and anticancer and antimicrobial potential of Juglans regia L. leaves. Molecules, 28(4), 1989. https://doi.org/10.3390/molecules28041989
  24. Welsh, J. (2013). Animal models for studying prevention and treatment of breast cancer. In Animal models for the study of human disease (pp. 997 1018). Academic Press. https://doi.org/10.1016/B978-0-12-415894-8.00040-3
  25. Cragg, G.M., Boyd, M.R., Khanna, R., Kneller, R., Mays, T.D., Mazan, K.D.,. & Sausville, E.A. (1999). International collaboration in drug discovery and development: the NCI experience. Pure and Applied Chemistry, 71(9), 1619-1633. https://doi.org/10.1351/pac199971091619
  26. Elouafy, Y., El Yadini, A., El Moudden, H., Harhar, H., Alshahrani, M.M., Awadh, A.A.A.,. & Tabyaoui, M. (2022). Influence of the extraction method on the quality and chemical composition of walnut (Juglans regia L.) oil. Molecules, 27(22), 7681. https://doi.org/10.3390/molecules27227681
  27. Jenab, M., Ferrari, P., Slimani, N., Norat, T., Casagrande, C., Overad, K.,. & Riboli, E. (2004). Association of nut and seed intake with colorectal cancer risk in the European Prospective Investigation into Cancer and Nutrition. Cancer Epidemiology Biomarkers & Prevention, 13(10), 1595-1603. https://doi.org/10.1158/1055-9965.EPI-04-0161.
  28. Pereira, J.A., Oliveira, I., Sousa, A., Valentão, P., Andrade, P.B., Ferreira, I.C.,. & Estevinho, L. (2007). Walnut (Juglans regia L.) leaves: Phenolic compounds, antibacterial activity and antioxidant potential of different cultivars. Food and Chemical Toxicology, 45(11), 2287 2295. https://doi.org/10.1016/j.fct.2007.06.004
  29. Verhoeckx, K., Cotter, P., López-Expósito, I., Kleiveland, C., Lea, T., Mackie, A.,. & Wichers, H. (2015). The impact of food bioactives on health: in vitro and ex vivo models. https://doi.org/10.1007/978-3-319-16104-4