Görkem KISMALI, Funda KOSOVA, Özlem TEMİZ ARPACI, Ercüment ÖLMEZ, Mustafa ARISOY, Zeki ARI, Feyzan ÖZDEL KURT, İbrahim TUĞLU

Anticancer effects of a newly-synthesized benzoxazole-derived 5-amino-2- [P-bromophenyl]-benzoxazole in breast cancer cell lines

We investigated the anticancer potential of 5-amino-2-[p-bromophenyl]-benzoxazole [BB] which is a new benzoxazole derivative in different breast cancer cell lines. BB was applied to estrogen receptor positive [ER+] MCF-7 and receptor negative [ER-] MDA-MB cell lines and its effects were determined by histopathological examinations, viability test [MTT], immunocytochemistry assays [Tunnel, VEGF and eNOS]. Moreover, its effects on apoptozis-related proteins such as Apaf-1, cytochrome C, caspase-3, bcl-2 and NF-κB were examined by western blot. Histopathological examinations showed that BB killed many cells for both cancer cell line while the cells lost their adhesion in MCF-7 and lost their adhesion and epiteloid morphology in MDA-MB. MTT analyses demonstrated that BB caused a clear dose depended toxic effect for both cell types. BB application resulted in an increase labeled apoptotic cells after Tunnel staining which was more obvious for MDA-MB compared to that of MCF-7. VEGF staining was decreased after BB application in both cell lines. The decrease of VEGF staining was clearer for MDA-MB compared to that of MCF-7. The status of oxidative stress was shown by eNOS immunocytochemistry which was increase after BB application in both cell lines. The levels of Apaf-1 and bcl-2 were found to be similar while caspase 9 and NF-κB levels were decreased compared to the control group after BB application for both cell lines. On the other hand, cytochrome C levels were slightly increased in MCF-7 cells while this increase was found very obvious in MDA-MB cell lines in BB groups. In conclusion, BB which is a new benzoxazole derivative have shown significant anticancer effects by increasing apoptosis and decreasing angiogenesis in MCF-7 or MDA-MB breast cancer cell lines. These effects of BB seem to be more prominent for [ER-] MDA-MB cell lines which mimic more aggressive type of breast cancer. These results suggest that BB may be useful to treat [ER-] breast cancers and may be added to treatment protocols.

PDF

___

1. LeSaygıner A. Treatment for angiogenesis, metastasis and tumor cell enzymes in cancer,5. İç hastalıkları kongresi, İstanbul Üniversitesi Onkoloji Enstitüsü, 2004;10-25.
2. He X, Dong Y, Wah Wu C, et al. MicroRNA–218 inhibits cell cycle progression and promotes apoptosis in colon cancer by downregulating BMI1 polycomb ring finger oncogene. Molecular Med. 2012;18:1491–8.
3. Akpınar G. Investigation of apoliprotein e [apo e] gene polypherism in colon cancer. Thesis, Kocaeli Unviversity; 2006;25-42.
4. Duranyıldız D,Oğuz H, Çamlıca H,et al. Serum BCL-2 levels in patients with malignant melanoma. Turkish J Oncol. 2004;19:131-3.
5. Goldstein JC, Waterhouse NJ, Juin P, et al. The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Nat Cell Biol. 2000;2:156-16215.
6. Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide- ranging implications in tissue kinetics. Br J Cancer. 1972;26:239- 57.
7. M Prudhomme, J Guyot, G Jeminet, Semi-synthesis of A23187 (calcimycin) analogs. IV. Cation carrier properties in mitochondria of analogs with modified benzoxazole rings. Antimicrobial activity, J Antibiot. 1986;39:934-7.
8. Vinsova J, Horak V, Buchta V, Kaustová J. Highly lipophilic benzoxazoles with potential antibacterial activity. Molecules. 2005;10:783-90.
9. Rida SM, Ashour FA, El-Hawash SAM, et al. Synthesis of some novel benzoxazole derivatives as anticancer, anti-HIV-1 and antimicrobial agents. J Med Chem. 2005;40:949-60.
10. Kumar D, Jacob MR, Reynolds MB, et.al. Synthesis and evaluation of anticancer benzoxazoles and benzimidazoles related to UK-1, Bioorg Med Chem. 2002;10:3997-4004.
11. Varga A, Akı-Şener E, Yalcın I, et al. Induction of apoptosis and necrosis by resistance modifiers benzazoles and benzoxazines on tumour cell line mouse lymphoma L5718 Mdr+cellsIn Vivo. 2005;19:1087-100.
12. Temiz-Arpaci O, Yildiz I, Ozkan S, et.al. Synthesis and biological activity of some new benzoxazoles. Eur J Med Chem. 2008;43:1423-31.
13. Kiessing S, Rockensuess KD, Varga A, Composition useful for the chemosensibiling and simultaneous apoptosis induction of multidrug resistance of tumor cells and bacteria, comprise substituteddisiloxane compound in the combination with benzoxazole derivatives. Patent De. 2010,102008027361 [A1].
14. Tekıner-Gulbas B, Temız-Arpacı O, Yıldız I, et al. 3D-QSAR study on heterocyclic topoisomerase II inhibitors using CoMSIA. SAR QSAR Environl Res. 2006;17:121–32.
15. Haansuu JP, Klika KD, Söderholm PP, et al. Isolation and biological activity of frankiamide, J Ind Microbiol. Biotechnol. 2001;27:62-6.
16. Chan M, Chang MC, González R, et al. Outcomes of estrogen receptor negative and progesterone receptor positive breast cancer. PLoS One, 2015;10:e0132449.
17. Clayton CC. Effect of certain benzimidazoles and related compounds upon azo dye destruction by liver homogenates[23789]. Proceed Soc Exp Biol Medic. 1958;98:510-2.
18. Schulze W, Gutsche W, Jungstand W. Zusammenhänge zwischen chemischer struktur und biologischer wirksamkeit bei azomethinen mit sticksofflostgruppen am ehrlich-ascitestumor der weissen Maus. Arzneimittelforschung. 1965;15:1235-38.
19. Sato S, Kajıura T, Mısato N, et al. AJI9561, A new cytotoxic benzoxazole derivative produced by streptomyces sp. J Antibio. 2001;54:102-4.
20. Lage H, Akı-Sener E, Yalcın I. High antineoplastic activity of newheterocyclic compounds in cancer cells with resistance against classical DNATopoisomerase II-targeting drugs. Int J Cancer. 2006;119:213-20.
21. Costales A, Mathur M, Ramurthy S, et.al. Shafer2- Amino-7-substituted benzoxazole analogs as potent RSK2 inhibitors Bioorg. Med Chem Lett. 2014;24:1592-96.
22. Kosova F, Arı Z, The relationship between prostate cancer and apoptozis. Journal of Clinical and Experimental Investigations. 2011;2:124-31.
23. Uluoğlu Ö. Basic pathology, 1990; 4. Baskı:855-870.
24. Özkaya AB. Investigation of the Effect of [-]-Epigallocatechin-3-gallate, the Active Ingredient of Green Tea, on Apoptosis in HCT-116 Colon Cancer Cell Line. Izmir, Dokuz Eylul University; 2008;1-50.
25. Kerr JF, Wyllie AH. Currie A.R. Apoptozis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972;26:239- 57.
26. Adams JM and Cory S. The Bcl–2 apoptotic switch in cancer development and therapy Oncogene. 2007;26:1324-37.
27. Takayama S, Reed JC and Homma S. Heat-shock proteins as regulators of apoptozis. Oncogene. 2003;22:9041–47.
28. Watanapokasin R, Jarinthanan F, Nakamura Y, et.al. Effects of α-mangostin on apoptozis induction of human colon cancer. World J Gastroenterol, 2011;17:2086-95.
29. Diaz GD, Li Q, Dashwood HR. Caspase-8 and apoptozis-inducing factor mediate a cytochrome c-independent pathway of apoptozis in human colon cancer cells induced by the dietary phytochemical chlorophyllin. Cancer Research. 2003;63:1254-61.
30. Manns J, Daubrawa M, Driessen S, et.al. Triggering of a novel intrinsic apoptozis pathway by the kinase inhibitor staurosporine: activation of caspase-9 in the absence of Apaf-1. The FASEB Journal. 2011;25:3250-61.
31. Watson AJM. Apoptosis and colorectal cancer. Gut 2004;53:1701-09.
32. Liu H, Jıang X, Zhang M, et.al. Association of CASP9, CASP10 gene polymorphisms and tea drinking with colorectal cancer risk in the Han Chinese population. Journal of Zhejiang University-Science Biomedicine & Biotechnology. 2013;14:47-57.
33. Saleh HA, Jacks H. Correlation of bcl-2 oncoprotein immunohistochemical expressionwith proliferation index and histopathologic parameters in colorectal neoplasia. Pathol Oncol Res. 1999;5:4.
34. Wright KM, Smith MI, Farrag L, and Deshmukh M. Chromatin modification of Apaf-1 restricts the apoptotic pathway in mature neurons. Journal Cell Biol. 2007;179:825-32.
35. Sakamoto K, Maeda S. Targeting NF-kappaB for colorectal cancer. Expert Opin Ther Targets 2010;14:593-601.
36. Wu Y, Deng J, Rychahou P.G, et.al. Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Cancer Cell. 2009;15:416-28.
37. Morgan MJ, Liu ZG. Crosstalk of reactive oxygen species and NF- kappaB signaling. Cell Res. 2011;21:103-15.
38. Zhang Y, Guo J, Zeng L, et.al. Tert- butyl-2[4,5-dihydrogen-4,4,5,5- tetramethyl-3-O-1H-imidazole-3-cationic- 1-oxy l-2-pyrrolidine-1- carboxylic ester displays novel cytotoxicity through reactive oxygen species-mediated oxidative damage in MCF-7 and MDA-MB-231 cells. Chem Biol Interact, 2011;192:287-97.
39. Funda Kosova, Ozlem Temiz Arpaci, Ercument Olmez, et.al. Synthesis and the effect of a novel benzoxazole compound on breast cancer cell line. Med Science. 2019;8:186-91.