YABANMERSİNİ (VACCİNİUM MYRTİLLUS L.) MEYVE ÖZÜ'NÜN MİDE KANSERİ, AGS HÜCRE HATTI ÜZERİNDE ANTİOKSİDAN AKTİVİTESİ VE ANTİ-KANSER ETKİLERİ

Öz

Amaç: Vaccinium myrtillus L. meyvesi Türkiye’de gıda olarak tüketilmektedir. Bu araştırma V. myrtillus metanol ekstresinin antioksidan ve sitotoksisite aktivitelerini ve mide kanser hücresindeki potansiyel etkilerini belirlemeyi amaçlamıştır.
Gereç ve Yöntem: V. myrtillus meyvesi metanol ekstresinin antioksidan aktiviteleri spektrofotometrik olarak ABTS+ ve DPPH• yöntemleriyle gerçekleştirilmiştir. Fitokimyasal analiz HPLC yöntemi ile araştırılmıştır. V. myrtillus metanol ekstresinin mide kanseri hücre hattı AGS üzerindeki sitotoksik etkisi, Cell Titer-Glo testi ile ölçülmüştür. Ek olarak, anti-kanser etkinlik analizi için fibroblast benzeri insan mezenkimal kök hücre hattı sağlıklı kontrol hücreleri olarak kullanılmıştır. Morfolojik değişiklikler, faz kontrast mikroskobu kullanılarak incelenmiştir.
Sonuç ve Tartışma: V. myrtillus meyve metanol ekstresinde ABTS+ ve DPPH• testleri ile sırasıyla 0.1413 ve 0.0439 mg/mL IC50 değerleri hesaplanmıştır. Bir antosiyanin bileşiği olarak malvidin-3-O-glukozit HPLC yardımıyla tespit edilmiştir. Sitotoksite analizi, farklı konsantrasyonlar (0.5-10 mg/ml) arasında 2 mg/ml V. myrtillus meyve metanol ekstresinin, sağlıklı MSC hücrelerini korurken AGS mide kanseri hücrelerinin canlılığını azalttığını göstermiştir. Bu veriler, Yaban mersini özünün çeşitli kanser hücre dizileri üzerinde daha fazla analiz edilmesini ve ekstresindeki potansiyel aktif maddelerin belirlenmesini önermektedir.

Anahtar Kelimeler

• Anti-kanser
• antioksidan aktivite
• mide kanseri
• Vaccinium myrtillus
• yaban mersini

ANTIOXIDANT ACTIVITY AND ANTI-CANCER EFFECTS OF BILBERRY (VACCINIUM MYRTILLUS L.) FRUIT EXTRACT ON GASTRIC CANCER, AGS CELL LINE

Öz

Objective: Vaccinium myrtillus L. fruits are consumed as food. This research was aimed to evaluate V. myrtillus methanol extract antioxidant and cytotoxic activities and determine its anti-cancer potential to further study against gastric cancer.
Material and Method: V. myrtillus fruit (Bilberry) methanol extract was examined for its antioxidant activities by ABTS•+ and DPPH• assays. The phytochemical analysis of the extract was studied by HPLC method. The cytotoxic effect of V. myrtillus fruit methanol extract on gastric cancer cell line AGS was measured by Cell Titer-Glo assay. Additionally, as healthy control, fibroblast like human mesenchymal stem cell line was used for testing anti-cancer efficacy.
Result and Discussion: V. myrtillus fruit methanol extract showed 0.1413 and 0.0439 mg/mL IC50 values as antioxidant activity by ABTS•+ and DPPH• assays, respectively. Malvidin-3-O-Glucoside was detected as an anthocyanin compound by HPLC method. Cytotoxicity analysis showed that among different concentrations (0.5-10 mg/ml), the most significantly, 2 mg/ml of Bilberry extract treatment decreased the viability of AGS gastric cancer cells while sparing healthy MSC cells. This data suggests the further analysis of Bilberry extract on several cancer cell lines as well as the determination of a potential active substance in the extract.

Anahtar Kelimeler

• Anti-cancer
• antioxidant activity
• bilberry
• gastric cancer
• Vaccinium myrtillus

Kaynakça

1. 1. Baytop, T. (1999). Türkiyede Bitkiler ile Tedavi, Nobel Tıp Kitabevleri, Istanbul.
2. 2. Bujor, O.C., Le Bourvellec, C., Volf, I., Popa, V.I., Dufour, C. (2016). Seasonal variations of the phenolic constituents in bilberry (Vaccinium myrtillus L.) leaves, stems and fruits, and their antioxidant activity. Food Chemistry, 213, 58–68. [CrossRef]
3. 3. Güner, A. (2012). Türkiye Bitkileri Listesi (Damarlı Bitkiler), Nezahat Gökyiğit Botanik Bahçesi Yayınları, Istanbul.
4. 4. Rouanet, J.M., Décordé, K., Rio, D.D. , Auger, C., Borges, G., Cristol, J.P., Lean M.E.J., Crozier, A. (2010). Berry juices, teas, antioxidants and the prevention of atherosclerosis in hamsters. Food Chemistry, 118(2), 266–271. [CrossRef]
5. 5. Ochkur, O., Del Bubba, M., Orlandini, S., Kravchenko, I., Koshovyi, O., Checchini, L., Bohachik, J. (2019). Study on the Polyphenols and Antocyanins Content In the Vaccinium myrtillus L. fruits harvested in different regions of Ukraine. Electronic Archive of the National Pharmaceutical University, 285–287. [CrossRef]
6. 6. Benvenuti, S., Brighenti, V., Pellati, F. (2018). High-performance liquid chromatography for the analytical characterization of anthocyanins in Vaccinium myrtillus L.(bilberry) fruit and food products. Analytical and Bioanalytical Chemistry, 410(15), 3559-3571. [CrossRef]
7. 7. Nyström, J., Östman, U.B., Mossing, T., Hed, L., Geladi, P. (2020). Solvent and Rotating Bed Reactor Extraction with One- and Two-Phase Solvents Applied to Bilberries (Vaccinium myrtillus) for Isolating Valuable Antioxidants. Food Analytical Methods, 13(4), 933–941. [CrossRef]
8. 8. Madhavi, D.L., Bomser, J., Smith, M.A.L., Singletary, K. (1998). Isolation of bioactive constituents from Vaccinium myrtillus (bilberry) fruits and cell cultures. Plant Science, 131(1), 95–103. [CrossRef] 9. Colak, N., Primetta, A.K., Riihinen, K.R., Jaakola, L., Grúz, J., Strnad, M., Torun, H., Ayaz, F.A. (2017). Phenolic compounds and antioxidant capacity in different-colored and non-pigmented berries of bilberry (Vaccinium myrtillus L.). Food Bioscience, 20, 67–78. [CrossRef]

9. 10. Machado, A.P.D.F., Pereira, A.L.D., Barbero, G.F., Martínez, J. (2017). Recovery of anthocyanins from residues of Rubus fruticosus, Vaccinium myrtillus and Eugenia brasiliensis by ultrasound assisted extraction, pressurized liquid extraction and their combination. Food Chemistry, 231, 1–10. [CrossRef]
10. 11. Satoh, Y., Ishihara, K. (2020). Investigation of the antimicrobial activity of Bilberry (Vaccinium myrtillus L.) extract against periodontopathic bacteria. Journal of Oral Biosciences, 62(2), 169–174. [CrossRef]
11. 12. Carmen Crespo, M., Visioli, F. (2017). A brief review of blue-and bilberries’ potential to curb cardio-metabolic perturbations: focus on diabetes. Current Pharmaceutical Design, 983–988. [CrossRef]
12. 13. Georgieva, A., Stefanova, R., Krastev, K. (2020). Exploration of The Impact of The Extraction Parameter on Biological Active Substances in The Extracts from Dried Fruit Black Blueberry (Vaccinium myrtillus L.). Trakia Journal of Sciences, 1(18), 59–68. [CrossRef]
13. 14. Matsunaga, N., Tsuruma, K., Shimazawa, M., Yokota, S., Hara, H. (2010). Matsunaga, N., Tsuruma, K., Shimazawa, M., Yokota, S., Hara, H. (2010). Inhibition of cholinesterase and amyloid-beta aggregation by resveratrol oligomers from Vitis amurensis. Phytotherapy Research, 24(S1), 42–47. [CrossRef]
14. 15. Puupponen-Pimiä, R., Nohynek, L., Alakomi, H.L., Oksman-Caldentey, K.M. (2005). Bioactive berry compounds - Novel tools against human pathogens. Applied Microbiology and Biotechnology, 67(1), 8–18. [CrossRef]
15. 16. Nguyen, V., Tang, J., Oroudjev, E., Lee, C. J., Marasigan, C., Wilson, L., Ayoub, G. (2010). Cytotoxic effects of bilberry extract on MCF7-GFP-tubulin breast cancer cells. Journal of Medicinal food, 13(2), 278–285. [CrossRef]
16. 17. Kang, S.Y., Seeram, N.P., Nair, M.G., Bourquin, L.D. (2003). Tart cherry anthocyanins inhibit tumor development in ApcMin mice and reduce proliferation of human colon cancer cells. Cancer Letters, 194(1), 13–19. [CrossRef]
17. 18. Ekşi, S., Aktura, S.Ş., Ejder, N., Şahin, K. (2019). In Vitro Cytotoxic Effect of Various Fruits on Human Cervical Carcinoma Cells. Journal of Biotechnology and Strategic Health Research, 2(3), 111–116. [CrossRef]
18. 19. Parkin, D.M. (2006). The global health burden of infection-associated cancers in the year 2002. International Journal of Cancer, 118(12), 3030–3044. [CrossRef]
19. 20. Cravo, M., Fidalgo, C., Garrido, R., Rodrigues, T., Luz, G., Palmela, C., Santos, M., Lopes, F. Maio, R. (2015). Towards curative therapy in gastric cancer: Faraway, so close! World Journal of Gastroenterology, 21(41), 11609–11620. [CrossRef]
20. 21. Duthie, S.J. (2007). Berry phytochemicals, genomic stability and cancer: Evidence for chemoprotection at several stages in the carcinogenic process. Molecular Nutrition and Food Research, 51(6), 665–674. [CrossRef]
21. 22. Blois, M.S. (1958). Antioxidant determinations by the use of a stable free radical, Nature Publishing Group, p.1199-1200. [CrossRef]
22. 23. Spanos, G.A., Wrolstad, R.E. (1990). Influence of processing and storage on the phenolic composition of Thompson Seedless grape juice. Journal of Agricultural and Food Chemistry, 38(7), 1565-1571. [CrossRef]
23. 24. Cesa, S., Carradori, S., Bellagamba, G., Locatelli, M., Casadei, M.A., Masci, A., Paolicelli, P. (2017). Evaluation of processing effects on anthocyanin content and colour modifications of blueberry (Vaccinium spp.) extracts: Comparison between HPLC-DAD and CIELAB analyses. Food Chemistry, 232, 114-123. [CrossRef]
24. 25. Martín‐Aragón, S., Basabe, B., Benedí, J.M., Villar, A.M. (1998). Antioxidant action of Vaccinium myrtillus L. Phytotherapy Research, 12(S1), S104-S106. [CrossRef]
25. 26. Beccaro, G. L., Mellano, M. G., Botta, R., Chiabrando, V., Bounous, G. (2006). Phenolic and anthocyanin content and antioxidant activity in fruits of bilberry (Vaccinium myrtillus L.) and of highbush blueberry (V.corymbosum L.) cultivars in North Western Italy, Acta Horticulture, 715, 553-557. [CrossRef]
26. 27. Dragana, M.V., Miroslav, R.P., Branka, B.R.G., Olgica, D.S., Sava, M.V., Ljiljana, R.Č. (2013). Antibacterial and antioxidant activities of bilberry (Vaccinium myrtillus L.) in vitro. African Journal of Microbiology Research, 7(45), 5130-5136. [CrossRef]
27. 28. Burdulis, D., Šarkinas, A., Jasutiene, I., Stackevičiene, E., Nikolajevas, L., Janulis, V. (2009). Comparative study of anthocyanin composition, antimicrobial and antioxidant activity in bilberry (Vaccinium myrtillus L.) and blueberry (Vaccinium corymbosum L.) fruits. Acta Poloniae Pharmaceutica - Drug Research, 66(4), 399-408.
28. 29. Güder, A., Gür, M., Engin, M.S. (2018). Antidiabetic and Antioxidant Properties of Bilberry ( Vaccinium myrtillus Linn.) Fruit and Their Chemical Composition. Journal of Agricultural Science and Technology, 17(2), 401-414.
29. 30. Ancillotti, C., Ciofi, L., Pucci, D., Sagona, E., Giordani, E., Biricolti, S., Gori M., Petrucci W.A., Giardi, F., Bartoletti, R., Chiuminatto, U., Orlandini, S., Mosti, S., Del Bubba, M. (2016). Polyphenolic profiles and antioxidant and antiradical activity of Italian berries from Vaccinium myrtillus L. and Vaccinium uliginosum L. subsp. gaultherioides (Bigelow) S.B. Young. Food Chemistry, 204, 176-184. [CrossRef]
30. 31. Martz, F., Jaakola, L., Julkunen-Tiitto, R., Stark, S. (2010). Phenolic Composition and Antioxidant Capacity of Bilberry (Vaccinium myrtillus) Leaves in Northern Europe Following Foliar Development and Along Environmental Gradients. Journal of Chemical Ecology, 36(9), 1017-1028. [CrossRef]
31. 32. Chu, W., Cheung, S.C.M., Lau, R.A.W., Benzie, I.F.F. (2011). Bilberry (Vaccinium myrtillus L.), Benzie, I.F.F., Wachtel-Galor, S. (Eds.), In Herbal Medicine: Biomolecular and Clinical Aspects, Herbal Medicine: Biomolecular and Clinical Aspects, Boca Raton (FL): CRC Press/Taylor, Francis, pp. 55-71. [CrossRef]
32. 33. Thibado, S., Thornthwaite, J., Ballard, T., Goodman, B. (2017). Anticancer effects of Bilberry anthocyanins compared with NutraNanoSphere encapsulated Bilberry anthocyanins. Molecular and Clinical Oncology, 8(2), 330-335. [CrossRef]
33. 34. Upton, R. (2001). Bilberry Fruit: Vaccinium Myrtillus L. : Standards of Analysis, Quality Control, and Therapeutics, American Herbal Pharmacopoeia and Therapeutic Compendium.
34. 35. Khoo, H.E., Azlan, A., Tang, S.T., Lim, S.M. (2017). Anthocyanidins and anthocyanins: colored pigments as food, pharmaceutical ingredients, and the potential health benefits. Food & Nutrition Research, 61(1), 1361779. [CrossRef]
35. 36. Li, D., Wang, P., Luo, Y., Zhao, M., Chen, F. (2017). Health benefits of anthocyanins and molecularmechanisms: Update from recent decade. Critical Reviews in Food Science and Nutrition, 8(57), 1729-1741. [CrossRef]
36. 37. Erlund, I., Koli, R., Alfthan, G., Marniemi, J., Puukka, P., Mustonen, P., Mattila, P., Jula, A. (2008). Favorable effects of berry consumption on platelet function, blood pressure, and HDL cholesterol. American Journal of Clinical Nutrition, 87(2), 323-331. [CrossRef]
37. 38. Alhosin, M., León-González, A.J., Dandache, I., Lelay, A., Rashid, S.K., Kevers, C., Pincemail, J., Fornecker, L.M., Mauvieux, L., Herbrecht, R., Schini-Kerth, V.B. (2015). Bilberry extract (Antho 50) selectively induces redox-sensitive caspase 3-related apoptosis in chronic lymphocytic leukemia cells by targeting the Bcl-2/Bad pathway. Scientific Reports, 5, 8996. [CrossRef]
38. 39. Huang, W., Zhu, Y., Li, C., Sui, Z., Min, W. (2016). Effect of blueberry anthocyanins malvidin and glycosides on the antioxidant properties in endothelial cells. Oxidative Medicine and Cellular Longevity, 2016, 1591803. [CrossRef]
39. 40. Baba, A.B., Nivetha, R., Chattopadhyay, I., Nagini, S. (2017). Blueberry and malvidin inhibit cell cycle progression and induce mitochondrial-mediated apoptosis by abrogating the JAK/STAT-3 signalling pathway. Food and Chemical Toxicology, 109, 534-543. [CrossRef]
40. 41. Oliveira, H., Wu, N., Zhang, Q., Wang, J., Oliveira, J., De Freitas, V., Mateus, N., He, J., Fernandes, I. (2016). Bioavailability studies and anticancer properties of malvidin based anthocyanins, pyranoanthocyanins and non-oxonium derivatives. Food and Function, 7(5), 2462-2468. [CrossRef]
41. 42. Wang, Y., Lin, J., Tian, J., Si, X., Jiao, X., Zhang, W., Gong, E., Li, B. (2019). 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, 67(2), 625-636. [CrossRef]