EFFECT OF 1,2-EPOXY-3[3-3[3,4-DIMETOXYI-PHENIL]-4H-1-BENZOPIRAN-4-ON] PROPANE (EPI) ON SIRTUIN-1 AND NUCLEAR FACTOR-κB EXPRESSION OF MAMMARY TUMORS INDUCED IN SPRAGUE DAWLEY RATS BY DMBA

https://doi.org/10.19106/JMedSci004803201602

Ayyub Harly Nurung(1*), Sri Herwiyanti(2), Dewi Kartikawati Paramita(3)

(1) Department of Histology and Cell Biology, Faculty of Medicine Universitas Gadjah Mada
(2) Department of Histology and Cell Biology, Faculty of Medicine Universitas Gadjah Mada
(3) Department of Histology and Cell Biology, Faculty of Medicine Universitas Gadjah Mada
(*) Corresponding Author

Abstract


ABSTRACT

The main factors contribute to breast cancer development is the combination of exogenous and endogenous factors. Endogenous factors include both SIRT1 and NF-kB. Exogenous factor used in this research is 7.12 dimethylbenz(a)anthracene (DMBA). 1,2-epoxy-3[3- 3[3,4-dimetoxy-phenil]-4h-1-benzophiran-4-on] propane (EPI) is a derivative of isoflavone generate from clove leaf oil. To examine the effect of EPI on SIRT1 and NF-kB expression in DMBA-induced Sprague Dawley (SD) rats, and the correlation between SIRT1 and NFkB expressions. Tissue generated form 35 Sprague Dawley female rats aged 2 weeks old were used in this study. Those rats were divided into 7 groups (5 rats/group), namely normal control group; corn oil group; DMBA group; EPI treated groups with 1 mg/kgBW (EPI I), 2 mg/kgBW (EPI II), and 4 mg/kgBW (EPI III), respectively; and doxorubicin group. EPI and doxorubicin were administered from 1st until 15th week, while DMBA were administered from 3rd until 9th week. The paraffin block was prepared from all breast organ of the rats that terminated at the end of week 15th. Examination of SIRT1 and NF-kB expression was performed using light microscope at 400x magnifications, after immunohistochemistry (IHC) staining. Expression level of SIRT1 and NF-kB were analyzed using IHC-profiler plug-in in ImageJ software. SIRT1 and NF-kB expression in EPI treated groups were not significantly different with the one in Doxorubicin group, but lower than DMBA group (p=0.000). There was a positive correlation between SIRT1 and NF-kB expression (p=0.001; r=0.773) in EPI-treated group. EPI was able to prevent an increasing of SIRT1 and NF-kB expression in SD model breast cancer that induced with DMBA. There is a positive correlation between SIRT1 and NF-kB expression in EPI-treated SD rats that were induced by DMBA


Keywords


Isoflavone, SIRT1 expression, NF-kB expression, breast cancer, EPI, DMBA, Sprague Dawley rats

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References

McDonald M, Herts RP, Lowenthal SW. The Burden of Cancer USA: Pfizer Medical Division; 2008. KEMENKES RI. InfoDATIN, STOP KANKER: Kementerian Kesehatan RI Pusat Data dan Informasi; 2015. Abdulkareem I, Zunmi I. Review of hormonal treatment of breast cancer. Niger. J. Clin. Pract. 2012; 15(1). Elangovan S, Ramachandran S, Venkatesan N, Ananth S, Gnana-Prakasam JP, Martin PM, et al. SIRT1 is essential for oncogenic signalling by estrogen/estrogen alfa in breast cancer. Cancer Res. 2011; 21(71): p. 6654-6664. Vaziri H, Dessain S, Eaton EN, Imai S, Frye AR, Pandita T, et al. hSIR2(SIRT1) functions as an NAD-dependent p53 deacetylase. Cell. 2001; 107: p. 149-159. Dai J, Wang Z, Sun D, Lin R, Wang SQ. SIRT1 interacts with p73 and suppresses p73-dependent transcriptional activity. J. Cell. Physiol. 2007; 210(1): p. 161-166. Wong S, Weber J. Deacetylation of the retinoblastoma tumour suppressor protein by SIRT1. Biochem. J. 2007;(407): p. 451-460. Kuo SJ, Lin Hy, Chien Sy, Chen Dr. SIRT1 suppresses breast cancer growth through downregulation of Bcl-2 protein. Oncol. Rep. 2013; 30: p. 125-130. Sas L, Lardon F, Vermeulen PB, Hauspy J, Dam PV, Pauwels P, et al. The Interaction between ER and Nf-kappaB in Resistance to Endocrine Therapy. Breast Cancer Research. 2012; 14: 212. Biswas DK, Shi Q, Baily S, Stricland I, Ghosh S, Pardee AB, et al. Nf-κB activation in human breast cancer specimens and its role in proliferation and apoptosis. PNAS. 2004; 101: p. 10137-10142. Hairil AA, Matsjeh S, Anwar C. Sintesis Senyawa Isoflavon dari Minyak Daun Cengkeh dan Uji Aktivitas Kanker secara in vitro. Disertasi. Yogyakarta: Universitas Gadjah Mada; 2010. Herwiyanti S. Potensi Senyawa 1,2-Epoksi-3[3-(3,4-dimetoksifenil)-4H-benzopiran-4on]propana sebagai Antikanker Kajian In Vitro Akitvitas Molekuler Sel Kanker Payudara T47D dan MCF-7 serta Aktivitas Molekuler In vivo pada Hewan Model yang Diinduksi 7,12-Dimetilbenz (a). Disertasi. Yogyakarta: Universitas Gadjah Mada; 2015. Varghese F, Bukhari AB, Malhotra R, De A. IHC Profiler: An Open Source Plugin for the Quantitative Evaluation and Automated Scoring of Immunohistochemistry Images of Human Tissue Samples. PLos ONE. 2014; 9(5): p. 1-11. Siegel R, Ma J, Zou Z, Jemal A. Cancer Statistic 2014. Ca Cancer J Clin. 2014; 64: p. 9-29. Androutsopoulos VP, Tsatsakis AM, Spandidos DA. Cytochrome P450 CYP1A1: Wider Roles in Cancer Progression and Prevention. BMC Cancer. 2009; 9(187): p. 1-17. Ohtake F, Takeyama Ki, Matsumoto T, Kitagawa H, Yamamoto Y, Nohara K, et al. Modulation of Oestrogen Receptor Signalling by Association with The Activated Dioxin Receptor. Nature. 2003; 423. Elangovan S, Ramachandran S, Venkatesan N, Ananth S, Prakasam J, Martin PM, et al. SIRT1 is Essential for Oncogenic Signaling by Estrogen/Estrogen Receptor α in Breast Cancer. Cancer Res. 2011; 71(21). Padauleng N, Purnomosari D, Herwiyanti S, Harjadi , Irianawati , Widyarini S. The Relationship Between Sirtuin 1 (SIRT1) Expression and Tumor SIze, Proliferating Cell Nuclear Antigen (PCNA) Expression and Histological Grading in Rat Breast Carcinoma Induced by dimethylbenz(alfa)anthracene (DMBA). Tesis. Yogyakarta: Universitas Gadjah Mada; 2013 Desember. Chen W, Wang D, Yen R, Luo J, Gu W, Baylin S. Tumor suppressor HIC1 directly regulates SIRT1 to modulate p53-dependent DNA-damage responses. Cell. 2005;(123): p. 437-448. Holloway RK, Barbieri A, Malyarchuk S, Saxena M, Kurepa NA, Mehl CM, et al. SIRT1 positively regulates breast cancer associated human aromatase (CYP19A1) Expression. Mol. Endocrinol. 2013; 27(3): p. 480-490. Kikuno N, Shiina H, Urakami S, Kawamoto K, Hirata H, Tanaka Y, et al. Genistein Mediated Histone Acetylation and Demethylatio Activates Tumor Suppressor Genes in Prostate Cancer Cells. Int. J. Cancer. 2008; 123: p. 552-560. Rasbach KA, Schnellmann RG. Isoflavones Promote Mitochondrial Biogenesis. JPET. 2008; 325: p. 536-543. Rahman SA, Shaban N, Haggag A, Awad D, Bassiouny A, Talaat I. Inhibition of Nf-κB, Bcl-2 and COX-2 GEne Expression by an Extract of Eruca Sativa Seeds during Rat Mammary Gland Carcinogenesis. Asian Pac J Cancer Prev. 2015; 16(18): p. 8411-8418. Currier N, Solomon SE, Demicco E, Chang DLF, Farago M, Ying H, et al. Oncogenic Signaling Pathways Activated in DMBA-Induced Mouse Mammary Tumors. Toxicologic Pathology. 2005; 33: p. 726-737. Shostak K, Chariot A. Nf-κB, Stem Cell and Breast Cancer: The Links Get Stronger. Breast Cancer Res. 2011; 13(214). Escarcega R, Fuentes-Alexandro S, Garcia-Carrasco M, Gatica A, Zamora A. The Transcription Factor Nuclear Factor-kappa B and Cancer. Clin Oncol (R Coll Radiol). 2007; 19(2): p. 154-161. Davis J, Kucuk O, Sarkar F. Genistein inhibits NF-kappaB activation in prostate cancer cells. Nutr Cancer. 1999; 35: p. 167-174. Baxa DM, Yoshimura FK. Genistein Reduces NF-kappaB in T Lymphoma Cells via a Caspase-Mediated Cleavage of I-Kappa-B-Alpha. Biochem Pharmacol. 2003; 66: p. 1009-1018. Tabary O, Escotte S, Coutil JP, Hubert D, Dusser D, Puchelle E, et al. Genistein Inhibits Constitutive and Inducible NFkappaB Activation and Decreases IL-8 Production bt Human Cystic Fibrosis Bronchial Gland Cells. Am J Pathol. 1999; 155: p. 473-481. Messina MJ, Wood EC. Soy isoflavones, estroge therapy, and breast cancer risk: analysis and commentary. Nutr. J. 2008; 7(17). Speirs V, Walkers RA. New Perspectives Into The Biological and Clinical Relevance of Oestrogen Receptors in The Human Breast. J Pathol. 2007; 499-506: 211. Kuiper GG, Lemmen JG, Carlsson B, Corton JC, Safe SH, Saag PTV, et al. Interaction of Estrogen Chemicals and Phytoestogens with Estrogen Receptor Beta. Endocrinology. 2013; 139. Xing D, Oparil S, Yu H, Gong K, Feng W, Black J, et al. Estrogen Modulates NFkB Signalling by Enhancing IkBa Levels and Blocking p65 Binding at the Promoters of Iflammatory Genes via Estrogen Receptor-B. PLoS ONE. 2012; VII(6): p. 1-10. Shertzer HG, Puga A, Chang Cy, Smith P, Nebert DW, Setchell KDR, et al. Inhibition of CYP1A1 Enzyme Activity in Mouse Hepatoma Cell Culture by Soybean Isoflavones. Chemico-Biological Interaction. 1999; 123: 31049. Robert DW, Doerge DR, Churchwell MI, Costa GG, Marques MM, Tolleson WH. Inhibiton of Extrahepatic Human Cytochromes P450 1A1 and 1B1 by Metabolism of Isoflavones Found in Trifolium pratense (Red Clover). J. Agric. Food Chem. 2004; 52(21): p. 6623-6632.



DOI: https://doi.org/10.19106/JMedSci004803201602

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