Cytotoxic and Pro-apoptotic Effects of Malvidin Anthocyanin in Berry Fruits on Colon Cancer

Öz

Aim: Increasing consumer consciousness elevated the consumption of fresh fruits and vegetables. Berries are among the most consumed fruits. The colorful nature of berries comes from the anthocyanins, a subgroup of flavonoids. The six anthocyanins in berries are cyanidin, pelargonidin, peonidin, delphinidin, petunidin, and malvidin, which gives blue color and is the least abundant (7%). Anti-carcinogenic and pro-apoptotic effects of malvidin have been studied insufficiently in literature due possibly to its small presence in berries. Method: In this study, the cytotoxic and pro-apoptotic effects of malvidin was measured on HT-29 human colon cancer cell line. Malvidin (0, 25, 50, 75, and 100 μM) was applied on HT-29 cells for 24, 48, and 72 hours, and cell viability was measured using MTT and CellTiter-Glo methods, while apoptosis indicators were measured using Caspase-Glo 3/7 assay. Results: Malvidin has showed dose- and time-dependent cytotoxicity on the cells with 72-hour IC50 value as low as 62.22 μM. To find out whether malvidin induced cytotoxicity through apoptosis, Caspase-Glo 3/7 assay was used, which showed that 25, 50, 75, and 100 μM malvidin applications elevated the caspase 3/7 activation at 72 hours by 1.26, 1.47, 1.94, and 2.21 times, respectively. Conclusion: As a result, it has been determined that malvidin has cytotoxic effects on colon cancer cells and this occurs via the activation of apoptosis pathway. Therefore, inclusion of berry fruits rich in anthocyanins in diet may be beneficial against gastrointestinal malignancies.

Anahtar Kelimeler

• anthocyanins
• berries
• cell culture
• cell proliferation
• apoptosis

Üzümsü Meyvelerdeki Malvidin Antosiyanininin Kolon Kanseri Üzerindeki Sitotoksik ve Pro-apoptotik Etkileri

Öz

Giriş ve Amaç: Tüketici bilincinin artması, taze meyve ve sebze tüketimini yükseltmiştir. Meyveler arasında en çok tüketilenler arasında kırmızı-mor meyveler bulunmaktadır. Meyvelerin rengini, flavonoidlerin bir alt grubu olan antosiyaninler sağlar. Meyvelerde bulunan altı antosiyanin türü; siyanidin, pelargonidin, peonidin, delfinidin, petunidin ve mavimsi rengi veren ve en az miktarda bulunan (%7) malvidindir. Malvidinin anti-kanserojen ve pro-apoptotik etkileri, meyvelerdeki düşük oranı nedeniyle literatürde yeterince araştırılmamıştır. Gereç ve Yöntemler: Bu çalışmada malvidinin HT-29 insan kolon kanseri hücre hattı üzerindeki sitotoksik ve pro-apoptotik etkileri ölçülmüştür. Malvidin (0, 25, 50, 75 ve 100 μM), HT-29 hücrelerine 24, 48 ve 72 saat boyunca uygulanmış ve hücre canlılığı CellTiter-Glo ve MTT metodları ile ölçülürken, apoptoz göstergeleri Caspase-Glo 3/7 testi ile değerlendirilmiştir. Bulgular: Malvidin, hücreler üzerinde doz ve zamana bağlı sitotoksik etki göstermiş ve 72 saatlik IC50 değeri 62,22 μM olarak belirlenmiştir. Malvidinin sitotoksisitesinin apoptoz yoluyla olup olmadığını belirlemek için Caspase-Glo 3/7 testi kullanılmış ve 25, 50, 75 ve 100 μM malvidin uygulamalarının 72 saatte sırasıyla 1,26, 1,47, 1,94 ve 2,21 kat artışla kaspaz 3/7 aktivasyonunu yükselttiği görülmüştür. Sonuç: Malvidinin kolon kanseri hücreleri üzerinde sitotoksik etkiler gösterdiği ve bunun apoptoz yolunun aktive edilmesiyle gerçekleştiği belirlenmiştir. Bu nedenle, antosiyaninler açısından zengin meyvelerin diyete dahil edilmesinin, gastrointestinal malignitelere karşı faydalı olabileceği düşünülmektedir.

Anahtar Kelimeler

• antosiyaninler
• üzümsü meyveler
• hücre kültürü
• hücre büyümesi
• apoptoz

Kaynakça

1. 1. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA: A Cancer Journal for Clinicians 2023; 73(1): 17-48.
2. 2. Wu H, Dai Q, Shrubsole MJ, Ness RM, Schlundt D, Smalley WE, Chen H, Li M, Shyr Y, Zheng W. Fruit and vegetable intakes are associated with lower risk of colorectal adenomas. The Journal of Nutrition 2009; 139(2): 340-4.
3. 3. Nile SH, Park SW. Edible berries: Bioactive components and their effect on human health. Nutrition 2014; 30(2): 134-44. https://doi.org/10.1016/j.nut.2013.04.007
4. 4. Khoo HE, Azlan A, Tang ST, Lim SM. Anthocyanidins and anthocyanins: Colored pigments as food, pharmaceutical ingredients, and the potential health benefits. Food & Nutrition Research 2017; 61: 1361779. https://doi.org/10.1080/16546628.2017.1361779
5. 5. Norberto S, Silva S, Meireles M, Faria A, Pintado M, Calhau C. Blueberry anthocyanins in health promotion: A metabolic overview. Journal of Functional Foods 2013; 5(4): 1518-28. https://doi.org/10.1016/j.jff.2013.08.015
6. 6. Saulite L, Jekabsons K, Klavins M, Muceniece R, Riekstina U. Effects of malvidin, cyanidin and delphinidin on human adipose mesenchymal stem cell differentiation into adipocytes, chondrocytes and osteocytes. Phytomedicine 2019; 53: 86-95. https://doi.org/10.1016/j.phymed.2018.09.029
7. 7. Lin J, Tian J, Shu C, Cheng Z, Liu Y, Wang W, Liu R, Li B, Wang Y. Malvidin-3-galactoside from blueberry suppresses the growth and metastasis potential of hepatocellular carcinoma cell Huh-7 by regulating apoptosis and metastases pathways. Food Science and Human Wellness 2020; 9(2): 136-45. https://doi.org/10.1016/j.fshw.2020.02.004
8. 8. Lopez de las Hazas MC, Mosele JI, Macia A, Ludwig IA, Motilva MJ. Exploring the colonic metabolism of grape and strawberry anthocyanins and their in vitro apoptotic effects in HT-29 colon cancer cells. Journal of Agricultural and Food Chemistry 2017; 65(31): 6477-87. https://doi.org/10.1021/acs.jafc.6b04096

9. 9. Zhang Y, Vareed SK, Nair MG. Human tumor cell growth inhibition by nontoxic anthocyanidins, the pigments in fruits and vegetables. Life Sciences 2005; 76(13): 1465-72. https://doi.org/10.1016/j.lfs.2004.08.025
10. 10. Xu H, Zhang J, Huang H, Liu L, Sun Y. Malvidin induced anticancer activity in human colorectal HCT-116 cancer cells involves apoptosis, G2/M cell cycle arrest and upregulation of p21WAFI. Int. J. Clin. Exp. Med. 2018;1 1(3): 1734-41.
11. 11. Feng SH, Zhao B, Zhan X, Li RH, Yang Q, Wang SM, Li A. Quercetin-induced pyroptosis in colon cancer through NEK7-mediated NLRP3 inflammasome-GSDMD signaling pathway activation. American Journal of Cancer Research 2024; 14(3): 934.
12. 12. Saulite L, Jekabsons K, Klavins M, Muceniece R, Riekstina U. Effects of malvidin, cyanidin and delphinidin on human adipose mesenchymal stem cell differentiation into adipocytes, chondrocytes and osteocytes. Phytomedicine 2019; 53: 86-95. https://doi.org/10.1016/j.phymed.2018.09.029
13. 13. Lin J, Tian J, Shu C, Cheng Z, Liu Y, Wang W, Liu R, Li B, Wang Y. Malvidin-3-galactoside from blueberry suppresses the growth and metastasis potential of hepatocellular carcinoma cell Huh-7 by regulating apoptosis and metastases pathways. Food Science and Human Wellness 2020; 9(2): 136-45. https://doi.org/10.1016/j.fshw.2020.02.004
14. 14. Lopez de las Hazas MC, Mosele JI, Macia A, Ludwig IA, Motilva MJ. Exploring the colonic metabolism of grape and strawberry anthocyanins and their in vitro apoptotic effects in HT-29 colon cancer cells. Journal of Agricultural and Food Chemistry 2017; 65(31): 6477-87. https://doi.org/10.1021/acs.jafc.6b04096
15. 15. Zhang Y, Vareed SK, Nair MG. Human tumor cell growth inhibition by nontoxic anthocyanidins, the pigments in fruits and vegetables. Life Sciences 2005; 76(13): 1465-72. https://doi.org/10.1016/j.lfs.2004.08.025
16. 16. Cetin E, Boyacioglu O, Orenay-Boyacioglu S. An effective treatment approach of liposomally encapsulated metformin in colon cancer. Medical Oncology 2024; 41(4): 82.
17. 17. Boyacioglu O, Kara B, Can H, Yerci TN, Yilmaz S, Boyacioglu SO. Leaf hexane extracts of two Turkish fig (Ficus carica L.) cultivars show cytotoxic effects on a human prostate cancer cell line. Agric Food Sci Res 2019; 6(1): 66-70. 18. Orenay Boyacioglu S. Role of PTEN in modulating preventive effect of 3, 4-DHPEA against oxidative stress. Sağlık Bilimleri Dergisi 2018; 27(1): 48-54.
18. 19. Malinowski P, Skała K, Jabłońska-Trypuć A, Koronkiewicz A, Wołejko E, Wydro U, Świderski G, Lewandowski W. Comparison of the usefulness of MTT and CellTiterGlo tests applied for cytotoxicity evaluation of compounds from the group of polyphenols. Environmental Sciences Proceedings 2022; 18(1): 9.
19. 20. Boyacıoğlu O. Zeytin karasuyu fenoliklerinden 3,4-dihidroksifenil etanolün farklı prostat kanseri hücre hatlarındaki sitotoksik etkileri. Food and Health 2019; 5(2): 95-100.
20. 21. Xu H, Zhang J, Huang H, Liu L, Sun Y. Malvidin induced anticancer activity in human colorectal HCT-116 cancer cells involves apoptosis, G2/M cell cycle arrest and upregulation of p21WAFI. International Journal of Clinical and Experimental Medicine 2018; 11(3): 1734-1741.
21. 22. Sakthivel KM, Brindha D. Malvidin abrogates oxidative stress and inflammatory mediators to inhibit solid and ascitic tumor development in mice. Journal of Environmental Pathology, Toxicology and Oncology 2020; 39(3): 247-260.
22. 23. Fritz J, Kern M, Pahlke G, Vatter S, Marko D. Biological activities of malvidin, a red wine anthocyanidin. Molecular Nutrition & Food Research 2006; 50(4-5): 390-395
23. 24. Bars-Cortina D, Sakhawat A, Piñol-Felis C, Motilva MJ. Chemopreventive effects of anthocyanins on colorectal and breast cancer: A review. In Seminars in Cancer Biology 2022 (Vol. 81, pp. 241-258). Academic Press.
24. 25. Caponio GR, Cofano M, Lippolis T, Gigante I, De Nunzio V, Difonzo G, Noviello M, Tarricone L, Gambacorta G, Giannelli G, De Angelis M, Notarnicola M. Anti-proliferative and pro-apoptotic effects of digested aglianico grape pomace extract in human colorectal cancer cells. Molecules. 2022; 27(20): 6791.
25. 26. Shi N, Chen X, Chen T. Anthocyanins in colorectal cancer prevention review. Antioxidants 2021; 10(10): 1600.
26. 27. Hyun JW, Chung HS. Cyanidin and malvidin from Oryza sativa cv. Heugjinjubyeo mediate cytotoxicity against human monocytic leukemia cells by arrest of G2/M phase and induction of apoptosis. Journal of Agricultural and Food Chemistry 2004; 52(8): 2213-2217.
27. 28. Wang Y, Lin J, Tian J, Si X, Jiao X, Zhang W, Gong E, Li B. Blueberry malvidin-3-galactoside suppresses hepatocellular carcinoma by regulating apoptosis, proliferation, and metastasis pathways in vivo and in vitro. Journal of Agricultural and Food Chemistry 2018; 67(2): 625-636.
28. 29. Dhivya S, Khandelwal N, Abraham SK, Premkumar K. Impact of anthocyanidins on mitoxantrone-induced cytotoxicity and genotoxicity: an in vitro and in vivo analysis. Integrative Cancer Therapies 2016; 15(4): 525-534.
29. 30. Dahlawi H. Effect of malvidin on induction of apoptosis and inhibition of cell proliferation on myeloid and lymphoid leukemia. Sch J Appl Med Sci 2022; 10: 150-156.
30. 31. Ma Y, Li Y, Zhang H, Wang Y, Wu C, Huang W. Malvidin induces hepatic stellate cell apoptosis via the endoplasmic reticulum stress pathway and mitochondrial pathway. Food Science & Nutrition 2020; 8(9): 5095-5106.
31. 32. Yeh CT, Yen GC. Induction of apoptosis by the Anthocyanidins through regulation of Bcl-2 gene and activation of c-Jun N-terminal kinase cascade in hepatoma cells. Journal of Agricultural and Food Chemistry 2005; 53(5): 1740-1749.
32. 33. Shih PH, Yeh CT, Yen GC. Effects of anthocyanidin on the inhibition of proliferation and induction of apoptosis in human gastric adenocarcinoma cells. Food and Chemical Toxicology 2005; 43(10): 1557-1566.
33. 34. Kobori M. In vitro-screening for cancer-suppressive effect of food components. Japan Agricultural Research Quarterly: JARQ 2003; 37(3): 159-165.
34. 35. Ulukaya E, Ozdikicioglu F, Oral AY, Demirci M. The MTT assay yields a relatively lower result of growth inhibition than the ATP assay depending on the chemotherapeutic drugs tested. Toxicology In Vitro 2008; 22(1): 232-9.
35. 36. Ulukaya E, Colakogullari M, Wood EJ. Interference by anti-cancer chemotherapeutic agents in the MTT-tumor chemosensitivity assay. Chemotherapy 2004; 50(1): 43-50.