AHU SOYOCAK, DİDEM TURGUT COŞAN, AYŞE YELDA BAŞARAN, Hasan Veysi GÜNEŞ, İRFAN DEĞİRMENCİ

Evaluation of Bax protein in breast cancer cells treated with tannic acid

Amaç: Bir bitki polifenolü olan Tannik Asit (TA)’in antikarsinojenik, antioksidan, antimutajenik, antimikrobiyal, antiallerjik ve antiinflamatuar aktiviteleri vardır. Bununla birlikte şimdiye kadar, meme kanserinde TA’in antikanser aktivitesinden sorumlu kesin bir mekanizma henüz açıkça tanımlanamamıştır. Bu çalışmanın amacı, insan meme kanser hücre dizisinde (MCF-7) TA’in pro-apoptotik Bax proteini üzerine etkisini araştırmaktır. Gereç ve yöntem: Bu çalışmada meme kanser hücrelerine (MCF-7) çeşitli konsantrasyon (0, 25, 50 and 100 μM) ve saatlerde (24, 48 ve 72. saatlerde) TA uygulandı. Bu hücrelerdeki pro-apoptotik Bax protein yüzdesini belirlemek için immunohistokimyasal boyama yöntemi kullanıldı. Bulgular: Araştırmamızın sonunda, çeşitli dozlarda TA muamelesinden sonra kanser hücrelerindeki pro-apoptotik Bax proteini yüzdesinin zamana bağımlı olarak arttığı gözlemledik. Bu artışın özellikle 72. saat’te ve 25 μM’lük dozda en yüksek düzeyde olduğu görüldü. Sonuç: Bulgularımıza göre, TA’in Bax proteinini artırarak, meme kanseri hücrelerinin apoptoza gitmesini sağlayabileceğini düşünmekteyiz. Bu çalışmadaki sonucun desteklenebilmesi için TA’in apoptotik yoldaki diğer proteinlerle ilişkisinin ayrıntılı bir şekilde değerlendirilmesi gerekmektedir.

Tannik asit uygulanan meme kanseri hücrelerinde Bax proteinin değerlendirilmesi

Objectives: Tannic acid (TA), a plant polyphenol, is known to have anti-carcinogenic, anti-oxidant, anti-mutagenic, anti-microbial, anti-allergic, anti-inflammatory activities. However, a precise mechanism responsible for the anti-cancer activity of TA in breast cancer has not yet been clearly described. The aim of this study was to investigate the effect of TA on the pro-apoptotic Bax protein in a human breast cancer cell line (MCF-7). Materials and methods: In this study, TA in various concentrations (0, 25, 50 and 100 μM) were administrated at various time points (24th h, 48th h and 72nd h) to breast cancer cells (MCF-7). The percentages of pro-apoptotic Bax protein in these cells were determined by immunohistochemical staining. Results: At the completion of the study, percentages of the pro-apoptotic Bax protein in pro-apoptotic cancer cells were found to be increased in a time-dependent manner after exposure to various concentrations of TA. This increase was at highest level with the concentration of 25 μM at 72nd hour. Conclusion: Based on our results, we suggested that TA could induce apoptosis of breast cancer cells by increasing Bax protein. Further studies evaluating the relationship of TA with other proteins that have a role in apoptotic pathways are warranted to support the findings of this study.

PDF

___

1. Parton M, Dowsett M, Smith I. Studies of apoptosis in breast cancer. Brit Med J 2001;322:1528-1532.
2. Henry-Mowatt J, Dive C, Martinou JC, James D. Role of mitochondrial membrane permeabilization in apoptosis and cancer. Oncogene 2004;23:2850-2860.
3. Debatin K-M. Apoptosis pathways in cancer and cancer therapy. Cancer Immunol Immun 2004;53:153-159.
4. Jeffrey S. Armstrong Mitochondria: a target for cancer therapy. Brit J Pharmacol 2006;147:239-248.
5. Martínez-Arribas F, Martín-Garabato E, Zapardiel I, et al. Bax expression in untreated breast cancer: an immunocytometric study of 255 cases. Anticancer Res 2008;28:5a2595-2598.
6. Sturm I, Kohne CH, Wolff G, Petrowsky H, et al. Analysis of the p53/BAX pathway in colorectal cancer: low BAX is a negative prognostic factor in patients with resected liver metastases. J Clin Oncol 1999;17:1364–1374.
7. Sturm I, Petrowsky H, Volz R, et al. Analysis of p53/BAX/p16(ink4a/CDKN2) in esophageal squamous cell carcinoma: high BAX and p16(ink4a/CDKN2) identifies patients with good prognosis. J Clin Oncol 2001;19:2272–2281.
8. Selvendiran K, Koga H, Ueno T, et al. Luteolin promotes degradation in signal transducer and activator of transcription 3 in human hepatoma cells: an implication for the antitumor potential of flavonoids. Cancer Res 2006;66:4826-4834.
9. Kuo PL, Lin CC. Green tea constituent (-)-epigallocatechin-3-gallate inhibits Hep G2 cell proliferation and induces apoptosis through p53-dependent and Fas-mediated pathways. J Biomed Sci 2003;10:219-27.
10. Lee HJ, Wang, CJ, Kuo HC, Chou FP, Jean LF, Tseng TH. Induction apoptosis of luteolin in human hepatoma HepG2 cells involving mitochondria translocation of Bax/Bak and activation of JNK. Toxicol Appl Pharmacol 2005;203:124-131.
11. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12:564-582.
12. Taffetani S, Ueno Y, Meng F, et al. Tannic acid inhibits cholangiocyte proliferation after bile duct ligation via a cyclic adenosine 5’,3’- monophosphate-dependent pathway. Am J Pathol 2005;166:1671-1679.
13. Nam S, Smith DM, Dou QP. Tannic acid potently inhibits tumor cell proteasome activity, increases p27 and Bax expression, and induces G1 arrest and apoptosis. Cancer Epidem Biomar 2001;10:1083-1088.
14. Khan NS, Hadi SM. Structural features of tannic acid important for DNA degradation in presence of Cu (II). Mutagenesis 1998; 13: 271-274.
15. Sakagami H, Satoh K. Prooxidant action of two antioxidants: ascorbic acid and gallic acid; Anticancer Res 1997;17:221-224.
16. Liu KX, Yamamoto F, Sekine S, et al. Inhibitory effect of methylene blue-induced photooxidation on intimal thickening of vein graft. Ann Thorac Surg 1999;68:84-88.
17. Chena K-S, Hsiao Y-C, Kuo D-Y, et al. Tannic acid-induced apoptosis and -enhanced sensitivity to arsenic trioxide in human leukemia HL-60 cells. Leukemia Res 2009;33:297-307.
18. Li B, Dou QP. Bax degradation by the ubiquitin⁄proteasome-dependent pathway: involvement in tumor survival and progression. Proc Natl Acad Sci Usa 2000;97:3850-3855.
19. Bawadi HA, Bonsode RR, Trappey A, Truax RE, Losso JN. Inhibition of CaCo-2 colon, MCF-7 and Hs578T breast and Du145 prostatic cancer cell proliferation by water-soluble black bean condensed tannins. Cancer Lett 2005;218: 153-162.
20. Romero I, Paez A, Ferruelo A, Lujan M, Berenguer A. Polyphenols in red wine inhibit the proliferation and induce apoptosis of LNCaP cells. BJU Int 2002;89:950-954.