Silibinin'in Vitro Ortamda CCL-228-SW 480 Kolon Kanseri Hücre Hattı Üzerine Apoptotik Etkisinin Araştırılması

Amaç: Kolon kanseri, dünyada önemli bir morbidite ve mortalite nedenidir. Kolon kanseri, kansere bağlı ölümlerde erkeklerde akciğer ve prostat kanseri; kadınlarda meme ve akciğer kanserinden sonra üçüncü sırada yer almaktadır. Silibin, devedikeni (silybum marianum) bitkisinden elde edilen bir ekstraktır. Yapılan çalışmalarda silibinin deri, mide kanserlerinde kanser kemoprotektif ajan olduğu ve antikarsinojenik etki gösterdiği tespit edilmiş ayrıca meme, kolon, mesane kanseri hücre hatlarında hücre siklusunu durdurduğu ve apopitozisi tetiklediği akciğer tümörü büyümesini baskıladığı, gösterilmiştir. Çalışmamızda silibinin CCL-228-SW 480 kolon karsinoma hücre soyu üzerine etkisini monolayer kültür ortamında incelemeyi amaçladık. Materyal-Method: Çalışmamızda silibinin CCL-228SW 480 kolon karsinoma hücre soyu üzerine etkisini incelemek amacıyla 100 µM/ml doz silibin (ID50 inhibisyon dozu belirlenen) hücrelere uygulandı. Silibin (100 µM/ml) uygulandıktan sonra 24, 48 ve 72 saat süreler sonunda BRDU işaretleme indeksi ve aktif kaspaz-3 tayinine immünohistokimyasal yöntemle bakıldı. Bulgular: CCL-228- SW 480 kolon karsinoma hücrelerinde silibinin ID50 inhibisyon dozu 100 ?M/ml olarak belirlendi. Silibinin iki boyutlu kültür ortamında BRDU ile işaretlenen hücrelerin sayısını azalttığı, kaspaz-3 ile boyanan hücrelerin sayısını ise arttırdığı görüldü.Sonuç: Sonuç olarak silibin CCL-228-SW 480 kolon kanseri hücre soyunda hücre proliferasyonunu inhibe edici ve apoptozisi tetikleyici etki göstermiştir. Buradan yola çıkarak yapmış olduğumuz iki boyutlu kültür sonuçlarımızın daha sonra yapılacak silibin ve kanser çalışmalarına ışık tutacaği kanısındayız.

Investigation of Appoptotic Effect of Silibinin on CCL-228-SW 480 Column Cancer Cell Line: An In Vitro Study

Objective: Colon cancer is a major cause of morbidity and mortality in the world. It is third of cancer-related deaths after breast and lung cancer in women, lung cancer and prostate cancer in males. Silibin is an extract obtained from the plant of the thistle (silybum marianum). Studies have shown that silibin is a cancer chemoprotective agent in skin, stomach cancers, and shows anticarcinogenic effect. At the same time it's known that it suppresses cell cycle in breast, colon, bladder cancer cell lines and suppresses apoptosis-induced lung tumor growth. In our study, we aimed to investigate the effect of silibin on CCL-228-SW 480 colon carcinoma cell line in monolayer culture medium. Material-Method: ID50 inhibition was determined on the dose for silibin and after it was found 100 ?M/ml was applied to the cells to examine the effect of silibin on CCL228-SW 480 colon carcinoma cell line. Silibin (100 ?M/ml) was administered and after 24, 48 and 72 hours the BRDU marking index and active caspase-3 assay were determined by immunohistochemistry.Results: The ID50 inhibition dose of silibinin CCL-228-SW 480 colon carcinoma cells was determined to be 100 ?M/ml. Silibin decreased the number of cells marked with BRDU and increased the number of cells stained with caspase-3 in the two-dimensional culture.Conclusion: As a result, silibin CCL-228-SW 480 inhibited cell proliferation and apoptosis in colon cancer cell line. We believe that the results of our two-dimensional culture, which we have made here, will shed light on silibin and cancer studies to be done later.

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1. Pfister DG, Rubin DM, Elkin EB, Neill US, Duck E, Radzyner M, et al. Risk adjusting survival outcomes in hospitals that treat patients with cancer without information on cancer stage. JAMA oncology. 2015; 1(9): 1303-10.
2. Tomida C, Aibara K, Yamagishi N, Yano C, Nagano H, Abe T, et al. The malignant progression effects of regorafenib in human colon cancer cells. The Journal of Medical Investigation. 2015; 62(3.4): 195-8.
3. ttp://www.drahmetdobrucali.com/hastaliklar/kalin-barsakkanseri-kolon-kanseri-kolorektal-kanser/ (Erişim tarihi: Aralık 2017)
4. Webb AL, McCullough ML. Dietary lignans: potential role in cancer prevention. Nutrition and cancer. 2005; 51(2): 117- 31.
5. Acartürk R, Şifalı Bitkiler, Flora ve Sağlığımız. Ovak Yayınları, No: 1 Karşıyaka, İzmir, 1996.
6. Ding T, Tian S, Zhang Z, et al. Dtermination of active component in silymarin by RP-LC and LC/MS. J. Pharmacol. Biomed. Anal 2001; 26: 155-61
7. Kocaman N, Dabak DÖ. Hepatoprotektif bir ajan: Silymarin. Firat Med J 2015; 20(3): 128-132.
8. Sanchez-Sampedro MA, Pelaez R, Corchete P. An Arabinogalactan Protein Isolated from Medium of Cell Suspensions Cultures of Silybum marianum Gaernt. Carbohydrate Polymers, 2008; 71: 634-638.
9. Morazzoni P, Bombardelli E. Silybum marianum (Carduus marianus). Fitoterapia, 1995; 64: 3-42.
10. Fraschini F, Demartini G, Esposti D. Pharmacology of silymarin. Clinical Drug Investigation 2002; 22: 51-65.
11. Kren V, Walterova D. Silybin and Silymarin-new effects and applications. Biomed Papers 2005;149: 29-41.
12. Kang JS, Jeon YJ, Park SK, Yang KH, Kim HM. Protection against lipopolysaccharide-induced sepsis and inhibition of interleukin-1 and prostaglandin E2 synthesis by silymarin. Biochem Pharmacol 2004; 67: 175- 81.
13. Nencini C, Giorgi G, Michelli L. Protective effect of silymarin on oxidative stress in rat brain. Phytomedicine 2007; 14: 129-35
14. Soto C, Recoba R, Barron H, Alvarez C, Favari L. Silymarin increases antioxidant enzymes in alloxan-induced diabetes in rat pancreas. Comp Biochem Physiol 2003; 136: 205-12.
15. Vinh PQ, Sugie S, Tanaka T, Hara A, Yamada Y, Katayama M, et al. Chemopreventive effects of a flavonoid antioxidant silymarin on N-butyl-N-(4-hydroxybutyl) nitrosamineinduced urinary bladder carcinogenesis in male ICR mice. Jpn J Cancer Res. 2002; 93: 42-49.
16. Kohno H, Tanaka T, Kawabata K, Hirose Y, Sugie S, Tsuda H, et al. A Naturally Occurring Polyphenolic Antioxidant Flavonoid, Inhibits Azoxymethane-Induced Colon Carcinogenesis in Male F344 Rats. Int. J. Cancer, 2002; 101: 461-468.
17. Ahmad N, Gali H, Javed S, Agarwal R. Skin cancer chemoprotective effects of a flavonoid antioxidant silymarin are mediated via impairment of receptor tyrosine kinase signaling and perturbation in cell cycle progression, Biochem Biophys Res Commun. 1998; 248: 294-301.
18. Singh RP, Tyangi AK, Zhao J, Agarwal R. Silymarin inhibits growth and causes regression of established skin tumors in SENCAR mice via modulation of mitogen-activated protein kinases and induction of apoptosis, Carcinogenesis, 2002; 23: 499-510.
19. Dorai T, Aggarwal B. Role of Chemopreventive Agants in Cancer Therapy. Cancer Lett. 2004; 215: 129-140.
20. Zi X, GrassoAW, Kung H-J, Agarwal R. A flavonoid antioxidant Silibinin inhibits activation of erbB1 signaling, and induces cyclin-dependent kinase inhibitors, G1arrest and anti-carcinogenic effects in human prostate carcinoma DU145 cells. Cancer Res. 1998; 58: 1920-9.
21. Zi X, Feyes DK, Agarwal R. Anti-carcinogenic effect of a flavonoid antioxidant silymarin in human breast cancer cells MDA-MB 468: induction of G1 arrest through an increase in Cip1/p21concomitant with a decrease in kinase activity of CDKs and associated cyclins. Clin Cancer Res. 1998; 4: 1055-64.
22. Agarwal C, Singh RP, Dhanalakshm IS, et al.Silibinin upregulates the expression of cyclin-dependent kinase inhibitors and causes cell cycle arrest and apoptosis in human colon carcinoma HT-29 cells. Oncogene 2003; 22: 8271-82.
23. Tyagi A, Agarwal C, Harrison G, Glode LM, Agarwal R. Silibinin causes cell cycle arrest and apoptosis inhuman bladder transitional cell carcinoma cells by regulating CDKICDK-cyclin cascade, and caspase 3 and PARP cleavages. Carcinogenesis 2004; 25: 1711-20.
24. Erarslan E, Yüksel İ, Haznedaroğlu S. Kolorektal Karsinogenez ve Metabolik Sendrom İlişkisi. Cumhuriyet Med J. 2012; 34: 380-5.
25. Erarslan E, Yüksel İ. Obezite ve Gastrointestinal Kanser İlişkisi. Yeni Tıp Dergisi. 2011; 28(4): 203-6.
26. Bernacchia R, Preti R, Vinci G. Chemical Composition and Health Benefits of Flaxseed. Austin J Nutri Food Sci. 2014; 2(8): 1045.
27. Borlu M. Lavaş Ekmeğine Farklı Seviyelerde Keten (linum usitatissimum) Tohumu Unu Katkılanmasının Hamur ve Ekmek Özellikleri Üzerine Etkisi, Omega 3, Omega 6 Yağ Asitleri ve Lignan Açısından Değişimin Belirlenmesi, (Yüksek Lisans tezi), Pamukkale Üniversitesi: 2009.
28. Hausott B, Greger H, Marian B. Naturally occurring lignans efficiently induce apoptosis in colorectal tumor cells. Journal of cancer research and clinical oncology. 2003;129(10):569-76.
29. Sezgin C. Kanserde Bitkilerle Tedavide Örnek Uygulamalar. Bitkilerle Tedavi Sempozyumu. Merkezefendi Geleneksel Tıp Derneği s:73, 5-6 Haziran 2010.
30. Fuentealba C, Figuerola F, Estévez AM, Bastías JM, Muñoz O. Bioaccessibility of lignans from flaxseed (linum usitatissimum) determined by singlebatch in vitro simulation of the digestive process. Journal of the science of food and agriculture. 2014; 94(9): 1729-38.
31. During A, Debouche C, Raas T, Larondelle Y. Among plant lignans, pinoresinol has the strongest antiinflammatory properties in human intestinal Caco-2 cells. The Journal of Nutrition. 2012; 142(10): 1798-805.
32. Bernacchia R, Preti R, Vinci G. Chemical composition and health benefits of flaxseed. Austin J Nutri Food Sci. 2014; 2(8): 1045.
33. Danbara N, Yurı T, Tsujıta-Kyutoku M, Tsukamoto R, Uehara N, Tsubura A. Enterolactone induces apoptosis and inhibits growth of Colo 201 human colon cancer cells both in vitro and in vivo. Anticancer research. 2005; MayJun;25(3B):2269-76.
34. Nesbitt PD. Mammalian lignan production from flaxseed, studies in vitro and in humans. Graduate Department of Nutritional Sciences University of Toronto, Master of Science, 1997.
35. Baytop T. Türkiye'de Bitkiler ile Tedavi Geçimişte ve Bugün, Nobel Tıp Kitapevleri, 1999; 142-44.
36. ZhaoJ, Agarwal R. Tissue distribution of silibinin, the major active constituent of silymarin, in mice and its association with enhancement of phase II enzymes: implations in cancer chemoprevention. Carcinogenesis; 1999; 20 (11): 2101-2108.
37. Syng-Ook L, Yun-Jeong J, Hyo Gwon I, Cheorl-Ho K, Young-Chae C, In-Seon L. Silibinin suppresses PMAinduced MMP-9 expression by blocking the AP-1 activation via MAPK signaling pathways in MCF-7 human breast carcinoma cells. Biochemical and Biophysical Research Communications, 2007; 354: 165-171
38. John J L, Wei C, Ke-Qin H. 'Effects and mechanisms of silibinin on human hepatoma cell lines' World Journal of Gastroenterology, 2007; 13(40): 5299-5305.