3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions

Park,Hongzoo; Lee,Dae-Sung; Yim,Mi-Jin; Choi,Yung  Hyun; Park,Saegwang; Seo,Su-Kil; Choi,Jung  Sik; Jang,Won  Hee; Yea,Sung  Su; Park,Won  Sun; Lee,Chang-Min; Jung,Won-Kyo; Choi,Il-Whan

doi:10.3892/ijmm.2015.2202

July-2015 Volume 36 Issue 1

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July-2015 Volume 36 Issue 1

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Article

3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions

Authors:
- Hongzoo Park
- Dae-Sung Lee
- Mi-Jin Yim
- Yung Hyun Choi
- Saegwang Park
- Su-Kil Seo
- Jung Sik Choi
- Won Hee Jang
- Sung Su Yea
- Won Sun Park
- Chang-Min Lee
- Won-Kyo Jung
- Il-Whan Choi
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Affiliations: Department of Urology, Kangwon National University Hospital, Kangwon National University School of Medicine, Chuncheon, Gangwon-do, Republic of Korea, Marine Biodiversity Institute of Korea, Seocheon, Chungcheongnam-do, Republic of Korea, Department of Biochemistry, College of Oriental Medicine, Dongeui University, Busan, Republic of Korea, Department of Microbiology, College of Medicine, Inje University, Busan, Republic of Korea, Department of Internal Medicine, Busan Paik Hospital, College of Medicine, Inje University, Busan, Republic of Korea, Department of Biochemistry, College of Medicine, Inje University, Busan, Republic of Korea, Department of Physiology, Kangwon National University School of Medicine, Chuncheon, Gangwon-do, Republic of Korea, Department of Molecular Microbiology and Immunology, Warren Alpert School of Medicine, Providence, RI, USA, Department of Biomedical Engineering and Center for Marine-Integrated Biomedical Technology (BK21 Plus), Pukyong National University, Busan, Republic of Korea
Pages: 301-308
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Published online on: May 5, 2015

https://doi.org/10.3892/ijmm.2015.2202
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Abstract

Oxidative stress in the retinal pigment epithelium (RPE) can lead to the pathological causes of age-related macular degeneration (AMD). Hypoxia induces oxidative damage in retinal pigment epithelial cells (RPE cells). In this study, we investigated the capacity of 3,3'-diindolylmethane (DIM) to reduce the expression of vascular endothelial growth factor (VEGF) under hypoxic conditions, as well as the molecular mechanisms involved. Human RPE cells (ARPE-19 cells) were treated with cobalt chloride (CoCl2, 200 µM) and/or DIM (10 and 20 µM). The production of VEGF was measured by enzyme-linked immunosorbent assay. The translocation of hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) was determined by western blot analysis. The binding activity of HIF-1α and NF-κB was analyzed by electrophoretic mobility shift assay. The phosphorylation levels of mitogen-activated protein kinases (MAPKs) were measured by western blot analysis. The levels of mitochondrial reactive oxygen species (ROS) were detected by fluorescence microplate assay. The results revealed that DIM significantly attenuated the CoCl2-induced expression of VEGF in the ARPE-19 cells. The CoCl2-induced translocation and activation of HIF-1α and NF-κB were also attenuated by treatment with DIM. In addition, DIM inhibited the CoCl2-induced activation of p38 MAPK in the ARPE-19 cells. Pre-treatment with YCG063, a mitochondrial ROS inhibitor, led to the downregulation of the CoCl2-induced production of VEGF by suppressing HIF-1α and NF-κB activity. Taken together, the findings of our study demonstrate that DIM inhibits the CoCl2-induced production of VEGF by suppressing mitochondrial ROS production, thus attenuating the activation of HIF-1α and p38 MAPK/NF-κB.

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1	Costagliola C, Agnifili L, Arcidiacono B, Duse S, Fasanella V, Mastropasqua R, Verolino M and Semeraro F: Systemic thromboembolic adverse events in patients treated with intravitreal anti-VEGF drugs for neovascular age-related macular degeneration. Expert Opin Biol Ther. 12:1299–1313. 2012. View Article : Google Scholar : PubMed/NCBI
2	Campochiaro PA: Ocular neovascularization. J Mol Med Berl. 91:311–321. 2013. View Article : Google Scholar : PubMed/NCBI
3	Simó R, Villarroel M, Corraliza L, Hernández C and Garcia-Ramírez M: The retinal pigment epithelium: Something more than a constituent of the blood-retinal barrier - implications for the pathogenesis of diabetic retinopathy. J Biomed Biotechnol. 2010:1907242010. View Article : Google Scholar
4	Yang XM, Wang YS, Zhang J, Li Y, Xu JF, Zhu J, Zhao W, Chu DK and Wiedemann P: Role of PI3K/Akt and MEK/ERK in mediating hypoxia-induced expression of HIF-1alpha and VEGF in laser-induced rat choroidal neovascularization. Invest Ophthalmol Vis Sci. 50:1873–1879. 2009. View Article : Google Scholar
5	Ke Q and Costa M: Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol. 70:1469–1480. 2006. View Article : Google Scholar : PubMed/NCBI
6	Banerjee S, Kong D, Wang Z, Bao B, Hillman GG and Sarkar FH: Attenuation of multi-targeted proliferation-linked signaling by 3,3′-diindolylmethane (DIM): From bench to clinic. Mutat Res. 728:47–66. 2011. View Article : Google Scholar : PubMed/NCBI
7	Brignall MS: Prevention and treatment of cancer with indole-3-carbinol. Altern Med Rev. 6:580–589. 2001.
8	Shertzer HG and Senft AP: The micronutrient indole-3-carbinol: Implications for disease and chemoprevention. Drug Metabol Drug Interact. 17:159–188. 2000. View Article : Google Scholar
9	Lee SH, Kim JS, Yamaguchi K, Eling TE and Baek SJ: Indole-3-carbinol and 3,3′-diindolylmethane induce expression of NAG-1 in a p53-independent manner. Biochem Biophys Res Commun. 328:63–69. 2005. View Article : Google Scholar : PubMed/NCBI
10	Kunimasa K, Kobayashi T, Kaji K and Ohta T: Antiangiogenic effects of indole-3-carbinol and 3,3′-diindolylmethane are associated with their differential regulation of ERK1/2 and Akt in tube-forming HUVEC. J Nutr. 140:1–6. 2010. View Article : Google Scholar
11	Yu BC, Lee DS, Bae SM, Jung WK, Chun JH, Urm SH, Lee DY, Heo SJ, Park SG, Seo SK, et al: The effect of cilostazol on the expression of matrix metalloproteinase-1 and type I procollagen in ultraviolet-irradiated human dermal fibroblasts. Life Sci. 92:282–288. 2013. View Article : Google Scholar : PubMed/NCBI
12	Kiriakidis S, Andreakos E, Monaco C, Foxwell B, Feldmann M and Paleolog E: VEGF expression in human macrophages is NF-kappaB-dependent: Studies using adenoviruses expressing the endogenous NF-kappaB inhibitor IkappaBalpha and a kinase-defective form of the IkappaB kinase 2. J Cell Sci. 116:665–674. 2003. View Article : Google Scholar : PubMed/NCBI
13	Sodhi A, Montaner S, Miyazaki H and Gutkind JS: MAPK and Akt act cooperatively but independently on hypoxia inducible factor-1alpha in rasV12 upregulation of VEGF. Biochem Biophys Res Commun. 287:292–300. 2001. View Article : Google Scholar : PubMed/NCBI
14	Vadlapatla RK, Vadlapudi AD, Pal D, Mukherji M and Mitra AK: Ritonavir inhibits HIF-1α-mediated VEGF expression in retinal pigment epithelial cells in vitro. Eye (Lond). 28:93–101. 2014. View Article : Google Scholar
15	Guo M, Song LP, Jiang Y, Liu W, Yu Y and Chen GQ: Hypoxia-mimetic agents desferrioxamine and cobalt chloride induce leukemic cell apoptosis through different hypoxia-inducible factor-1α independent mechanisms. Apoptosis. 11:67–77. 2006. View Article : Google Scholar
16	Ratcliffe PJ, O’Rourke JF, Maxwell PH and Pugh CW: Oxygen sensing, hypoxia-inducible factor-1 and the regulation of mammalian gene expression. J Exp Biol. 201:1153–1162. 1998.PubMed/NCBI
17	Ratcliffe PJ, Pugh CW and Maxwell PH: Targeting tumors through the HIF system. Nat Med. 6:1315–1316. 2000. View Article : Google Scholar : PubMed/NCBI
18	Fay J, Varoga D, Wruck CJ, Kurz B, Goldring MB and Pufe T: Reactive oxygen species induce expression of vascular endothelial growth factor in chondrocytes and human articular cartilage explants. Arthritis Res Ther. 8:R1892006. View Article : Google Scholar : PubMed/NCBI
19	Xia C, Meng Q, Liu LZ, Rojanasakul Y, Wang XR and Jiang BH: Reactive oxygen species regulate angiogenesis and tumor growth through vascular endothelial growth factor. Cancer Res. 67:10823–10830. 2007. View Article : Google Scholar : PubMed/NCBI
20	Cervellati F, Cervellati C, Romani A, Cremonini E, Sticozzi C, Belmonte G, Pessina F and Valacchi G: Hypoxia induces cell damage via oxidative stress in retinal epithelial cells. Free Radic Res. 48:303–312. 2014. View Article : Google Scholar
21	Gupta B, Chiang L, Chae K and Lee DH: Phenethyl isothiocyanate inhibits hypoxia-induced accumulation of HIF-1α and VEGF expression in human glioma cells. Food Chem. 141:1841–1846. 2013. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions

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