Apoptozisin doğal indüksiyonu: Althaea officinalis L.'nin meme kanseri tedavisindeki potansiyel rolü

Abstract

Meme kanserinin tedavisinde cerrahi, kemoterapi ve radyoterapi en sık kullanılan yöntemlerdir. Ancak, bu uygulamaların yan etkileri nedeniyle semptomlar oldukça agresif ve ağrılıdır. Bu nedenle bitkisel tedavi ön plana çıkmıştır. Antimikrobiyal, antiviral, antienflamatuar, antioksidan, öksürük kesici, immünomodülatör ve hipoglisemik biyoaktiviteleri ile dikkat çeken bir bitki olan Althaea officinalis çiçeklerinin sulu ekstresi üzerinde herhangi bir çalışma yapılmamıştır. Bu nedenle, bu çalışmada, Althaea officinalis çiçeklerinin sulu ekstresinin MCF-7 hücre hattında canlılık ve apoptozla ilişkili genler üzerindeki doz ve zamana bağlı etkilerini belirledik. Althaea officinalis L.'nin hücre canlılığı üzerindeki etkisini incelemek amacıyla, MCF-7 hücrelerine farklı konsantrasyonlarda Althaea officinalis L. uygulanmış ve farklı zaman aralıklarında (24, 48, 72 saat) MTT yöntemi ile analiz edilmiştir. Ek olarak, sitotoksik dozların hücre ölümü üzerindeki etkisini her saatte belirlemek için, apoptozla ilişkili genler olan Bcl-2, Bax ve kaspaz-3'ün gen ekspresyon seviyelerindeki değişimler qRT-PCR ile belirlendi. Sonuçlara göre, IC50 dozları 24, 48 ve 72. saatlerde sırasıyla 837,83 µg/mL, 1,51 mg/mL ve 235,84 µg/mL olarak belirlendi. Apoptotik süreçte rol oynayan Bcl-2, Bax ve Kaspaz-3 genlerinin mRNA ekspresyon düzeyleri qPCR ile analiz edildi. Bcl-2 gen ekspresyonunun azaldığı, Bax ve Kaspaz-3 ekspresyon düzeylerinin ise arttığı belirlendi. Althaea officinalis L.'nin kanser karşıtı özellikleri, sulu ekstresinin MCF-7 hücreleri üzerinde sitotoksik etkiye sahip olması ve apoptozu indüklemesiyle dikkat çekici hale gelmiştir.

Keywords

• Althaea officinalis L.
• meme kanseri
• MCF-7
• apoptozis

Natural induction of apoptosis: the potential role of Althaea officinalis L. in breast cancer treatment

Abstract

Surgery, chemotherapy, and radiotherapy are the most commonly used methods in the treatment of breast cancer. However, due to the side effects of these treatments, symptoms are quite aggressive and painful. Therefore, herbal treatments have become prominent. No studies have been conducted on the aqueous extract of Althaea officinalis L, a plant noted for its immunomodulatory, antioxidant, anti-inflammatory, antimicrobial, antitussive, antiviral, and hypoglycemic bioactivities. Therefore, in this study, we determined the dose- and time-dependent effects of the aqueous extract of Althaea officinalis flowers on viability and apoptosis-related genes in the MCF-7 cell line. To examine the effect of Althaea officinalis L. on cell viability, different concentrations of Althaea officinalis L. were applied to MCF-7 cells and analyzed at different time intervals (24, 48, and 72 hours) using the MTT method. Additionally, to determine the effect of cytotoxic doses on cell death at each hour, changes in gene expression levels of apoptosis-related genes Caspase-3, Bax and, Bcl-2 were determined by qRT-PCR. According to the results, the IC50 doses were determined to be 837.83 µg/mL, 1.51 mg/mL, and 235.84 µg/mL at 24, 48, and 72 hours, respectively. The mRNA expression levels of Caspase-3, Bax and, Bcl-2 genes involved in the apoptotic process were analyzed by qPCR. It was determined that Bcl-2 gene expression decreased, while Bax and Caspase-3 expression levels increased. The anti-cancer properties of Althaea officinalis L. became notable due to the cytotoxic effect of its aqueous extract on MCF-7 cells and its induction of apoptosis.

Keywords

• Althaea officinalis L
• breast cancer
• MCF-7
• apoptozis

References

1. Giaquinto, A. N., Sung, H., Newman, L. A., ve diğerleri, Breast cancer statistics 2024, CA: A Cancer Journal for Clinicians, 74, 6, 477-495, (2024).
2. Siegel, R. L., Giaquinto, A. N., Jemal, A., Cancer statistics, 2024, CA: A Cancer Journal for Clinicians, 74, 1, 12-49, (2024).
3. Rossi, R. E., Pericleous, M., Mandair, D., Whyand, T., Caplin, M. E., The role of dietary factors in prevention and progression of breast cancer, Anticancer Research, 34, 12, 6861-6875, (2014).
4. Nagykálnai, T., Landherr, L., Nagy, A. C., Vitamin D and breast cancer, Orvosi Hetilap, 155, 28, 1091-1096, (2014).
5. Ma, H., Carpenter, C. L., Sullivan‐Halley, J., Bernstein, L., The roles of herbal remedies in survival and quality of life among long-term breast cancer survivors, BMC Cancer, 11, 222, (2011).
6. Complementary and Alternative Medicine Doctors Alliance, The regulatory status of complementary and alternative medicine for medical doctors in Europe, Raport / Publication, (2010).
7. Omogbadegun, Z. O., Medicinal plants-based foods for breast cancer treatment: an ethnobotanical survey and digitization, International Journal of Medical Plants and Alternative Medicine, 1, 8, 137-163, (2013).
8. Al-Snafi, A. E., The pharmaceutical importance of Althaea officinalis and Althaea rosea: A review, International Journal of PharmTech Research, 5, 3, 1387-1385, (2013).

9. Shah, S. A., Akhtar, N., Akram, M., ve diğerleri, Pharmacological activity of Althaea officinalis L., Journal of Medicinal Plants Research, 5, 24, 5662-5666, (2011).
10. Hage-Sleiman, R., Mroueh, M., Daher, C. F., Pharmacological evaluation of aqueous extract of Althaea officinalis flower, Pharmaceutical Biology, 49, 3, 327-333, (2011).
11. Talebi, A., Karimi, A., Ouguerram, K., ve diğerleri, Lack of nephroprotective efficacy of Althaea officinalis flower extract, International Journal of Preventive Medicine, 5, 11, 1360, (2014).
12. Fallahpour, F., Banaee, M., Javadzade, N., Effects of dietary Althaea officinalis extract on common carp, International Journal of Advanced Biological and Biomedical Research, 2, 8, 2453-2460, (2014).
13. Husain, M., Wadud, A., Sofi, G., Perveen, S., Hafeez, K. A., Physicochemical standardization of mucilage of Althaea officinalis, Pharmacognosy Magazine, 15, 62, (2019).
14. Rownaghi, M., Niakousari, M., Assessing physicochemical characteristics of a shear-thinning polysaccharide mucilage extracted from marshmallow root (Althaea officinalis L.) by an ohmic heating system, International Journal of Biological Macromolecules, 277, 134274, (2024).
15. Farhat, C., Younes, H., Alyamani, O. A., ve diğerleri, Chemical characterization and biological evaluation of Althaea officinalis flower extract, Journal of Herbal Medicine, 34, 100575, (2022).
16. Bonaterra, G. A., Bronischewski, K., Hunold, P., ve diğerleri, Anti-inflammatory effects of Althaea officinalis root extract, Frontiers in Pharmacology, 11, 290, (2020).
17. Özdemir, F. A., Turan, M., Investigation of antimicrobial and anticancer activity of extracts obtained following UV application to Althaea officinalis L. callus cultures, International Journal of Secondary Metabolite, 10, 3, 425-436, (2023).
18. Baimakhanova, B., Sadanov, A., Ratnikova, I., Orasymbet, S., Shorabaev, E., Baimakhanova, G., ... & Turgumbayeva, A., Phytochemical compounds and biomedical properties of Althaea officinalis, Pharmacy Practice, 1886-3655, 23, 3, (2025).
19. Okuyan, D., Epidermal growth factor downregulates carbonic anhydrase III in colon cancer, Current Issues in Molecular Biology, 46, 11, 12994-13002, (2024).
20. Atalay, M. S., Avcıkurt, A. S., & Tosun, S. G., The Effects of Althaea officinalis L. Extract on Cell Viability, Invasion, and Apoptosis in Prostate Cancer, Journal of Uludağ University Medical Faculty, 51, 3, 457-465, (2025).
21. Poyrazlı, F., Okuyan, D., Köçkar, F., Türkoğlu, S. A., Hypoxic regulation of KLK4 gene, Cell Biochemistry and Biophysics, 82, 3, 2797-2812, (2024).
22. Livak, K. J., Schmittgen, T. D., Analysis of relative gene expression using the 2−ΔΔCT method, Methods, 25, 4, 402-408, (2001).
23. Girden, E. R., ANOVA: Repeated Measures, SAGE Publications, (1992).
24. Hockenbery, D., Nuñez, G., Milliman, C., Schreiber, R. D., Korsmeyer, S. J., Bcl-2 blocks programmed cell death, Nature, 348, 6299, 334-336, (1990).
25. Miyashita, T., Krajewski, S., Krajewska, M., ve diğerleri, p53 regulates bcl-2 and bax gene expression, Oncogene, 9, 6, 1799-1805, (1994).
26. Ponder, K. G., Boise, L. H., Caspase-3 prodomain regulates activation, Cell Death Discovery, 5, 1, 56, (2019).
27. Al-Snafi, A. E., Anticancer activity of plant phenolic compounds, Iraqi Journal of Cancer and Medical Genetics, 4, 2, 66-71, (2011).
28. Al-Snafi, A. E., Detoxification capacity of medicinal plants, Asian Journal of Pharmaceutical Science & Technology, 5, 4, 257-270, (2015).
29. Al-Snafi, A. E., Clinically tested medicinal plants, SMU Medical Journal, 3, 1, 99-128, (2016).
30. Blumenthal, M., Goldberg, A., Brinckmann, J., Herbal Medicine: Expanded Commission E Monographs, Integrative Medicine Communications, 519 s., (2000).
31. Gudej, J., Determination of flavonoids in Althaea officinalis, Farmacja Polska, 46, 5, 153-155, (1990).
32. Gudej, J., Flavonoids and phenolic acids from Althaea officinalis, Planta Medica, 57, 3, 284-285, (1991).
33. Bradley, P. R., British Herbal Compendium: Marshmallow Root, British Herbal Medicine Association, (1992).
34. Franz, G., Schleimpolysaccharide von Althaea officinalis, Planta Medica, 14, 1, 90-110, (1966).
35. Rosík, J., Kardošová, R., Toman, R., Capek, P., Characterization of Althaea officinalis mucilage, Československá Farmacie, 33, 68-71, (1984).
36. Ashtiyani, C., Yarmohammady, P., Hosseini, N., Ramazani, M., Effect of Althaea officinalis root extract on diabetic rats, Iranian Journal of Endocrinology and Metabolism, 17, 3, 238-250, (2015).
37. Tahmouzi, S., Nejat, M. R. S., Infertility therapy effects of Althaea officinalis polysaccharides, International Journal of Biological Macromolecules, 145, 777-787, (2020).
38. Zhang, Y., Kong, F., Zhang, L., Li, C., Zhang, R., Effect of Althaea officinalis on A549 cancer cells, Tropical Journal of Pharmaceutical Research, 15, 12, 2647-2652, (2016).
39. Boeker, I., Sendker, J., Stark, T., ve diğerleri, Cytoprotective effects of marshmallow roots, Zeitschrift für Phytotherapie, 33, Suppl 1, P06, (2012).
40. Tobyn, G., Denham, A., Whitelegg, M., Medical Herbs: Western Herbal Tradition, Churchill Livingstone, (2011).
41. Vannucci, L., Krizan, J., Sima, P., ve diğerleri, Immunostimulatory and antitumor effects of glucans, International Journal of Oncology, 43, 2, 357-364, (2013).
42. Chan, G. C. F., Chan, W. K., Sze, D. M. Y., Effects of β-glucan on immune and cancer cells, Journal of Hematology & Oncology, 2, 1, 25, (2009).
43. Hashemifesharaki, R., Xanthakis, E., Altintas, Z., Guo, Y., Gharibzahedi, S. M. T., Microwave-assisted extraction of marshmallow polysaccharides, Carbohydrate Polymers, 240, 116301, (2020).
44. Suganya, K. U., Govindaraju, K., Kumar, V. G., Karthick, V., Parthasarathy, K., Gold nanoparticles induce apoptosis in breast cancer, International Journal of Biological Macromolecules, 93, 1030-1040, (2016).
45. Jackson, C. L., Dreaden, T. M., Theobald, L. K., ve diğerleri, Pectin induces apoptosis in prostate cancer, Glycobiology, 17, 8, 805-819, (2007).
46. Hosseini Abari, A., Amini Rourani, H., Ghasemi, S. M., Kim, H., Kim, Y. G., Antioxidant and anticancer activity of galacturonic acids, Scientific Reports, 11, 1, 8491, (2021).
47. Wathoni, N., Shan, C. Y., Shan, W. Y., ve diğerleri, Pectin characterization from mangosteen rind, Heliyon, 5, 8, e02299, (2019).
48. Kadhum, H. H., Abd, A. H., Al-Shammari, A. M., HPLC analysis of Althaea officinalis flavonoids, AIP Conference Proceedings, 2123, 1, 020045, (2019).
49. Kuntz, S., Wenzel, U., Daniel, H., Effects of flavonoids on colon cancer cells, European Journal of Nutrition, 38, 3, 133-142, (1999).
50. Ramos, S., Dietary flavonoids and apoptosis in cancer, Journal of Nutritional Biochemistry, 18, 7, 427-442, (2007).
51. HemaIswarya, S., Doble, M., Natural product synergism in cancer, Phytotherapy Research, 20, 4, 239-249, (2006).
52. Hanasaki, Y., Ogawa, S., Fukui, S., Antioxidative effects of flavonoids, Free Radical Biology and Medicine, 16, 6, 845-850, (1994).
53. Korkina, L. G., Afanas’ev, I. B., Antioxidant properties of flavonoids, Advances in Pharmacology, 38, 151-163, (1996).
54. Kampa, M., Alexaki, V. I., Notas, G., ve diğerleri, Phenolic acids inhibit proliferation in T47D breast cancer cells, Breast Cancer Research, 6, 2, R63, (2003).
55. Anantharaju, P. G., Gowda, P. C., Vimalambike, M. G., Madhunapantula, S. V., An overview on the role of dietary phenolics for the treatment of cancers, Nutrition Journal, 15, 1, 99, (2016).
56. Moon, J. Y., Mosaddik, A., Kim, H., ve diğerleri, Guava leaf extract induces apoptosis in gastric cancer, Food Chemistry, 125, 2, 369-375, (2011).
57. Keating, E., Martel, F., Antimetabolic effects of polyphenols in breast cancer, Frontiers in Nutrition, 5, 25, (2018).
58. Li, C. L., Han, X. C., Zhang, H., Wu, J. S., Li, B., Scopoletin induces apoptosis in prostate cancer, Tropical Journal of Pharmaceutical Research, 14, 4, 611-617, (2015).
59. Manuele, M. G., Ferraro, G., Arcos, M. L. B., ve diğerleri, Immunomodulatory effect of scopoletin, Life Sciences, 79, 21, 2043-2048, (2006).
60. Demirel, I., Vumma, R., Mohlin, C., ve diğerleri, Nitric oxide induces IL-6 expression, American Journal of Nephrology, 36, 6, 524-530, (2012).
61. Zhao, Y., Liu, J., Liu, C., ve diğerleri, Anti-inflammatory effects of p-coumaric acid, Medicinal Chemistry, 6, 327-330, (2016).
62. Lu, M., Dai, Y., Xu, M., ve diğerleri, Caffeic acid attenuates inflammation via 14-3-3ζ, Inflammation, 40, 5, 1753-1760, (2017).
63. Navarrete, S., Alarcón, M., Palomo, I., Tomato extract reduces TNF-α and IL-1β expression, Molecules, 20, 8, 15319-15329, (2015).
64. Huang, L., Hou, L., Xue, H., Wang, C., Gallic acid inhibits inflammation via TLR4/NF-κB, Cellular & Molecular Immunology Journal, 32, 12, 1610-1614, (2016).
65. Ginwala, R., Bhavsar, R., Chigbu, D. G. I., Jain, P., Khan, Z. K., Role of flavonoids in inflammation, Antioxidants, 8, 2, 35, (2019).
66. Liang, Y. C., Huang, Y. T., Tsai, S. H., ve diğerleri, Apigenin inhibits COX-2 and iNOS, Carcinogenesis, 20, 10, 1945-1952, (1999).
67. Lee, H. N., Shin, S. A., Choo, G. S., ve diğerleri, Anti-inflammatory effect of quercetin, International Journal of Molecular Medicine, 41, 2, 888-898, (2018).
68. Hämäläinen, M., Nieminen, R., Vuorela, P., Heinonen, M., Moilanen, E., Anti-inflammatory effects of flavonoids, Mediators of Inflammation, 2007, 1-10, (2007).
69. Baguley, B. C., Leung, E., Phenotype heterogeneity in breast cancer cell lines, Breast Cancer—Carcinogenesis, Cell Growth and Signalling Pathways, bölüm, 245-256, (2011).
70. Shirazi, F. H., Zarghi, A., Kobarfard, F., ve diğerleri, Investigating breast cancer compounds, Breast Cancer—Focusing Tumor Microenvironment, Stem Cells and Metastasis, 85-102, (2011).
71. Strasser, A., Harris, A. W., Huang, D. C., Krammer, P. H., Cory, S., Bcl-2 and Fas regulate apoptosis pathways, EMBO Journal, 14, 24, 6136-6147, (1995).
72. Adams, J. M., Cory, S., The Bcl-2 protein family, Science, 281, 5381, 1322-1326, (1998).
73. Youle, R. J., Strasser, A., BCL-2 family mediates cell death, Nature Reviews Molecular Cell Biology, 9, 1, 47-59, (2008).
74. Czabotar, P. E., Lessene, G., Strasser, A., Adams, J. M., Control of apoptosis by BCL-2 proteins, Nature Reviews Molecular Cell Biology, 15, 1, 49-63, (2014).
75. Babacan, D., Apoptozis, Selçuk University Journal of Science Faculty, 49, 1, 1-10, (2023).