AVEMAR HT29 KOLON KANSERİ HÜCRELERİNDEKİ ANTİ-ANJİYOJENİK ETKİSİNİ COX-2 VE VEGF EKSPRESYONLARINI İNHİBE EDEREK GÖSTERİR

Abstract

Fermente buğday tohumu özütünden elde edilen Avemar, tamamen doğal, klinikte özellikle kemoterapi ve radyoterapi tedavisi gören kanser hastalarında diyet desteği olarak kullanılan non-toksik bir bileşiktir. Çalışmada, birçok biyolojik aktiviteye sahip olan bu ekstraktın insan kolon kanseri hücrelerinde anti-anjiyojenik özelliğinin olup olmadığının belirlenmesi amaçlandı. Bu amaçla, Avemar 800, 1600 ve 3200 µg/ml dozlarında HT-29 hücrelerine uygulandı ve sonrasında, en önemli anjiyogenez belirteçlerinden olan VEGF (Vasküler Endotel Büyüme Faktörü) ve Cox-2 (Siklooksijenaz-2) protein ve mRNA miktarlarındaki değişiklikler araştırıldı. Protein miktarlarındaki değişiklikler Elisa yöntemi ile, mRNA düzeylerindeki değişiklikler ise gerçek zamanlı polimeraz zincir reaksiyonu ile belirlendi. Avemar uygulanan hücrelerdeki VEGF ve Cox-2 protein ve mRNA düzeyleri, uygulanmayan HT-29 hücrelerine göre önemli derecede azalma gösterdi. Araştırma sonunda elde edilen veriler, Avemar’ın kolon kanseri hücreleri üzerinde anti-anjiyojenik etkisinin olabileceği yönünde yorumlandı ve tıp dünyasına kolon kanserinin anti-anjiyojenik tedavisinde aday bir ekstrakt olarak aktarıldı

Keywords

• Avemar, anjiyogenez, kolon kanseri, vasküler endotel büyüme faktörü, siklooksijenaz-2

Avemar Exhibits Anti-Angiogenic Effect Mediated By Inhibition of Cox-2 and Vegf Gene Expression on Ht29 Colon Carcinoma Cells

Abstract

Avemar, obtained from fermented wheat germ extract, is a completely natural and non-toxic compound which is clinically used in particular as diet support in cancer patients treated with chemotherapy and radiotherapy. The purpose of this study is to determine whether avemar has an anti-angiogenic effects on human colon cancer cells. For this aim, HT29 cells were exposed to 400, 800, 1600 and 3200 µg/ml concentrations of Avemar, subsequently, protein amounts and mRNA levels of VEGF (Vascular Endothelial Growth Factor) and Cox-2 (Cyclooxygenase-2), which are the most important angiogenic markers, were investigated. While the change in the amount of protein was measured by Elisa assay, the alterations at the mRNA levels were determined by real time PCR. Avemar exhibited significantly important decrease in the both of protein and mRNA levels of VEGF and Cox-2 genes on HT29 cells treated with avemar, compared to non-treated cells. The results of this study led us to interpret that Avemar might be an anti- angiogenic effect on colon cancer cells, and Avemar was ssuggested to medical community as a candidate extract for the therapy of colon cancer.

Keywords

• Avemar, Angiogenesis, Colon cancer, Vascular endothelial growth factor, Cyclooxygenase-2.

References

1. Mueller T, Voigt W. Fermented wheat germ extract- nutrional supplement or anticancer drug? Nutrition Journal 2011; 10: 89.
2. Saiko P, Ozsvar-Kozma M, Madlener S, et al. Avemar, a nontoxic fermented wheat germ extract, induces apoptosis and inhibits ribonucleotide reductase in human HL-60 promyelocytic leukemia cells. Cancer Letters 2007; 250: 323-328.
3. http://www.avemar.com/what_is_avemar
4. Comin-Anduix B, Boros LG, Marin S, et al. Fermented wheat germ extract inhibits glycolysis/ pentose cycle enzymes and induces apoptosis through Poly(ADP-riboz) polymerase activation in Jurkat T-cell leukemia tumor cells. The J Biol Chem 2002; 277 (29): 46408-46414.
5. Mueller T, Jordan K, Voigt W. Promising cytotoxic profile of fermented wheat germ extract (Avemar) in human cancer cell lines. JExp Clin Cancer Res 2011; 30: 42.
6. Telekes A, Kiss-Toth E, Nagy T, et al. Synergistic effect of Avemar on Proinflammatory cytokine production and Ras-mediated cell activation. Ann NY Acad Sci 2005; 1051: 515-528.
7. Fajka-Boja R, Hidvegi M, Shoenfeld Y, et al. Fermented wheat germ extract induces apoptosis and downregulation of major histocompatibility complex I proteins in tumour T and B cell lines. Int J Oncol 2002; 20: 563-570.
8. Telekes A, Hidvegi M. Avemar’s mechanism of action (2)- Immunological effects. Nögyogyaszati Onkologia 2001; 6: 40-41.

9. Boros LG, Nıchelatti M, Shoenfeld Y. Fermented wheat germ extract (Avemar) in the treatment of cancer and autoimmune diseases. Ann NY Acad Sci 2005; 1051: 529-542.
10. Hidvegi M, Raso E, Tömösközi-Farkas R, Lapis K, Szende B. Effect of MSC on the immune response of mice. Immunopharmacology 1999; 41: 183-186
11. Illmer C, Madlener S, Horvath Z, et al. Immunologic and biochemical effects of fermented wheat germ extract Avemar. Exp Biol Med 2004; 230: 144-149.
12. Du Bois RN, Abramson SB, Crofford L, et al. Cyclooxygenase in biology and disease. FASEB J 1998; 12: 1063-1073.
13. Shiff SJ, Shiprasad P, Santini DL. Cyclooxygenase inhibitors: drugs for cancer prevention. Curr Opin Pharmacol 2003; 3: 352-361.
14. Xiong B, Sun TJ, Hu WD, et al. Expression of cyclooxygenase-2 in colorectal cancer and its clinical significance. World J Gastroenterol 2005; 28; 11 (8): 1105-1108.
15. Zhou ZG, Wu XJ, LiL R, et al. A multivariate analysis of prognostic determinants for stage II and III colorectal cancer in 141 patients. Chin Med J (Engl) 2011; 124(14): 2132-2135.
16. Nakamoto RH, Uetake H, Iida S, et al. Correlations between cyclooxygenase expression and angiogenic factors in primary tumors and liver metastases in colorectal cancer. Jpn J Clin Oncol 2007; 37 (9): 679-685.
17. Kobayashi H, Sugihara K, Uetake H, et al. Messenger RNA expression of cox-2 and angiogenic factors in primary colorectal cancer and corresponding liver metastasis. Int J Oncol 2009; 34(4): 1147- 1153.
18. Kolev Y, Uetake H, Iida S, et al. Prognostic significance VEGF expression in correlation with Cox-2, microvessel density, and clinicopathological characteristics in human gastric carcinoma. Ann Surg Oncol 2007; 14(10): 2738-2747.
19. Yang Q, Ye ZY, Zhang JX, et al. Expression of matrix metalloproteinase-9 mRNA and vascular endothelial growth factor protein in gastric carcinoma and its relationship to its pathological features and prognosis. Anat Rec (Hoboken) 2010; 293(12): 2012-2019.
20. Noriyuki M, Sumi T, Zhi X, et al. Vascular endothelial growth factor, matrix metalloproteinases, and cyclooxygenase-2 influence prognosis of uterine cervical cancer in young women. Int J Oncol 2007; 31(3): 531-536.
21. Gaffney DK, Haslam D, Tsodikov A, et al. Epidermal Growth Factor Receptor (EGFR) and vaskular endothelial growth factor negativelly affect overall survival in carcinoma of the cervix treated with radiotherapy. Int J Radiat Oncol Biol Phys 2003; 56 (4): 922-928.
22. Byun JH, Lee MA, Roh SY, et al. Association between cyclooxygenase-2 and matrixmetalloproteinase-2 expression in non-small cell lung cancer. Jpn J Clin Oncol 2006; 36(5): 263-268.
23. Warren RS, Yuan H, Matli MR, Gillett NA, Ferrara N. Regulation by vascular endothelial growth factor of human colon cancer tumorigenesis in a Mouse model of experimental liver metastasis. J Clin Invest 1995; 95: 1789-1797.
24. Reinmuth N, Liu W, Fan F, et al. Blockade of insulin -like growth factor I receptor function inhibits growth and angiogenesis of colon cancer. Clin Cancer Res 2002; 8: 3259-3269.
25. Ciardiello F, Caputo R, Bianco R, et al. Inhibition of growth factor production and angiogenesis in human cancer cells by ZD1839 (Iressa), a selective epidermal growth factor receptor tyrosine kinase inhibitor. Clin Cancer Res 2001; 7: 1459-1465.
26. Takahashi Y, Bucana CD, Cleary KR, Ellis LM. p53, vessel count and vascular endothelial growth factor expression in human colon cancer. Int J Cancer 1998; 79: 34-38.
27. Rak J, Mitsuhashi Y, Bayko L, et al. Mutant ras oncogenes upregulate VEGF/VPF expression: implications for induction and inhibition of tumor angiogenesis. Cancer Res 1995; 55: 4575-4580.
28. Ellis LM, Staley CA, Liu W, et al. Down-regulation of vascular endothelial growth factor in a human colon carcinoma cell line transfected with an antisense expression vector specific for c-src. J Biol Chem 1998; 273: 1052-1057.
29. Tsujii M, Kawano S, Tsuji S, et al. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell 1998; 93: 705-716.
30. Tsujii M, Kawano S and DuBois RN. Cyclooxygenase -2 expression in human colon cancer cells increases metastatic potential. Proc Natl Acad Sci USA 1997; 94: 3336-3340.
31. Yang MD, Chang WS, Tsai CW, et al. Inhibitory effects of AVEMAR on proliferation and metastasis of oral cancer cells. Nutrition and Cancer 2016; 68(3): 473-480.
32. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta C(T)) method. Methods 2001; 25 (4), 402-408.
33. Jung YD, Nakano K, Liu W, Gallick GE and Ellis LM. Extracellular signal-regulated kinase activation is required for up-regulation of vascular endothelial growth factor by serum starvation in human colon carcinoma cells. Cancer Res 1999; 59, 4804-4807.
34. Hidvegi M, Raso E, Tomoskozi-Farkas R, et al. MSC, a new benzoquinone-containing natural product with antimetastatic effect. Cancer Biother Radiopharm 1999; 14: 277-289.
35. Szende B, Raso E, Hidvegi M, et al. A new benzoquinone-containing antimetastatic product. Orv Hetil 1998; 139: 2893-2897.
36. Hidvegi M, Raso E, Tomoskozi Farkas R, et al. Effect of MSC on the immune response of mice. Immunopharmacology 1999; 41: 183-186.
37. Jakab F, Shoenfeld Y, Balogh A, et al. A medical nutriment has supportive value in the treatment of colorectal cancer. Br J Cancer 2003; 89: 465-469.
38. Zhang JY, Xiao X, Dong Y, et al. Effect of fermented wheat germ extract with lactobacillus plantarum dy-1 on HT-29 cell proliferation and apoptosis. J Agric Food Chem 2015; 63: 2449-2457.
39. R. S. Kerbel, Tumor angiogenesis. N Engl J Med 2008; 358: 2039-2049.
40. Taleb H, Morris RK, Withycombe CE, et al. Date syrup derived polyphenols attenuate angiogenic responses and exhibits anti-inflammatory activity mediated by VEGF and COX-2 expression in endothelial cells. Nutrition Research 2016; doi: 10.1016/j.nutres.2016.02.010.
41. Fan TP, Yeh JC, Leung KW, et al. Angiogenesis: from plants to blood vessels. Trends Pharmacol Sci 2006; 27: 297-309
42. Angelo LS and Kurzrock R. Vascular Endothelial Growth Factor and Its Relationship to InflammatoryMediators. Clin Cancer Res 2007; 13(10): 2825 -2830.
43. Szende B, Marcsek Z, Kocsis Z, Tompa A. Effect of simultaneous administration of Avemar and cytostatic drugs on viability of cell cultures, growth of experimental tumors, and survival of tumor- bearing mice. Cancer Biother Radiopharm 2004; 19: 343-349.
44. Demidov LV, Manzjuk LV, Kharkevitch GY, Artamonova EV, Pirogova NA. Antimetastatic effect of Avemar in high-risk melanoma patients Int J Cancer 2002; 100 (Suppl 13): 408.
45. Hidvegi M, Moldvay J, Lapis K, Ajkay Z. Fermented wheat germ extract improves quality of life in lung cancer patients [English translation]. Medicus Anonymus/ Pulmono 2003; 11(Suppl 1):13-14.
46. Garami M, Schuler D, Babosa M, et al. Fermented wheat germ extract reduces chemotherapy induced febrile neutropenia in pediatric cancer patients. J Pediatr Hematol Oncol 2004; 26: 631-635.