Fenretinide reduces angiogenesis by downregulating CDH5, FOXM1 and eNOS genes and suppressing microRNA-10b

  • Elif Isil Yücel
  • Mehmet SahinEmail author
Original Article


Angiogenesis is a new vessel formation process that plays a role in various physiological and pathological conditions. This process is controlled by the balance between pro-angiogenic and anti-angiogenic mediators in the organism. Angiogenesis is needed for the growth and metastasis of solid tumors. Therefore, the anti-angiogenic treatment approach is seen as an interesting option in cancers. Fenretinide, a synthetic retinoic acid analog, is an effective agent on angiogenesis. In this study, we aimed to investigate the effects of the fenretinide on some miRNAs involving in angiogenesis process and on the expression of CDH5, FOXM1 and eNOS genes upregulated in angiogenesis. In addition, it was shown the effects of this agent on cell proliferation, cell migration and capillary-like tube formation. In our study, the data were analyzed using Kruskal–Wallis and Dunn’s test. Fenretinide applied to the cells for 24 and 48 h periods reduced cell proliferation (P < 0.001) and cell migration, and suppressed tube formation (P < 0.001) as a dose dependent manner. Endothelial cells were cultured in growth-inducing media containing a variety of growth factors such as VEGF, FGF, IGF and EGF. As a result of simultaneous PCR analysis, we found that angiogenesis-promoting miR-10b was effectively suppressed (P < 0.001) and interestingly angiogenesis-modulating miR-126 was slightly increased (P < 0.05), but other miRNAs, including miR-31, miR-21, miR-101, miR-340, miR-29c, miR-206 and miR-146a were not affected. Besides, a significant decrease was observed in the levels of some angiogenesis-inducing genes, CDH5 (P < 0.001), FOXM1 (P < 0.001) and eNOS (P < 0.01 and P < 0.001) in endothelial cells treated with fenretinide. Our results have shown that fenretinide exhibited anti-angiogenic activity through the down-regulation of CDH5, FOXM1 and eNOS genes, and suppression of miR-10b.


Fenretinide Angiogenesis CDH5 eNOS FOXM1 miRNA 



Funding for this study was provided by grants from Gaziantep University Scientific Research Project Unit (Project Number: TF.YLT.18.43). Also this study was supported by Gaziantep University Health Sciences Institute.

Compliance with ethical standards

Conflict of interest

We declare that all authors have no potential conflict of interest.


  1. 1.
    Sahin M, Sahin E, Gumuslu S (2009) Cyclooxygenase-2 in cancer and angiogenesis. Angiology 60(2):242–253. CrossRefPubMedGoogle Scholar
  2. 2.
    Sahin M, Sahin E, Gumuslu S, Erdogan A, Gultekin M (2010) DNA methylation or histone modification status in metastasis and angiogenesis-related genes: a new hypothesis on usage of DNMT inhibitors and S-adenosylmethionine for genome stability. Cancer Metastasis Rev 29(4):655–676. CrossRefPubMedGoogle Scholar
  3. 3.
    Bussolino F, Mantovani A, Persico G (1997) Molecular mechanisms of blood vessel formation. Trends Biochem Sci 22(7):251–256CrossRefGoogle Scholar
  4. 4.
    Du J, Yang Q, Luo L, Yang D (2017) C1qr and C1qrl redundantly regulate angiogenesis in zebrafish through controlling endothelial Cdh5. Biochem Biophys Res Commun 483(1):482–487. CrossRefPubMedGoogle Scholar
  5. 5.
    Gartel AL (2010) A new target for proteasome inhibitors: FoxM1. Expert Opin Investig Drugs 19(2):235–242. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Park S, Sorenson CM, Sheibani N (2015) PECAM-1 isoforms, eNOS and endoglin axis in regulation of angiogenesis. Clin Sci (Lond) 129(3):217–234. CrossRefGoogle Scholar
  7. 7.
    Al Tanoury Z, Piskunov A, Rochette-Egly C (2013) Vitamin A and retinoid signaling: genomic and nongenomic effects. J Lipid Res 54(7):1761–1775. CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Huang P, Chandra V, Rastinejad F (2014) Retinoic acid actions through mammalian nuclear receptors. Chem Rev 114(1):233–254. CrossRefPubMedGoogle Scholar
  9. 9.
    Sogno I, Vene R, Ferrari N, De Censi A, Imperatori A, Noonan DM, Tosetti F, Albini A (2010) Angioprevention with fenretinide: targeting angiogenesis in prevention and therapeutic strategies. Crit Rev Oncol Hematol 75(1):2–14. CrossRefPubMedGoogle Scholar
  10. 10.
    Anding AL, Jones JD, Newton MA, Curley RW Jr, Clagett-Dame M (2018) 4-HPR Is an endoplasmic reticulum stress aggravator and sensitizes breast cancer cells resistant to TRAIL/Apo2L. Anticancer Res 38(8):4403–4416. CrossRefPubMedGoogle Scholar
  11. 11.
    Anding AL, Chapman JS, Barnett DW, Curley RW Jr, Clagett-Dame M (2007) The unhydrolyzable fenretinide analogue 4-hydroxybenzylretinone induces the proapoptotic genes GADD153 (CHOP) and Bcl-2-binding component 3 (PUMA) and apoptosis that is caspase-dependent and independent of the retinoic acid receptor. Cancer Res 67(13):6270–6277. CrossRefPubMedGoogle Scholar
  12. 12.
    Ferrari N, Morini M, Pfeffer U, Minghelli S, Noonan DM, Albini A (2003) Inhibition of Kaposi's sarcoma in vivo by fenretinide. Clin Cancer Res 9(16 Pt 1):6020–6029PubMedGoogle Scholar
  13. 13.
    Ferrari N, Pfeffer U, Dell'Eva R, Ambrosini C, Noonan DM, Albini A (2005) The transforming growth factor-beta family members bone morphogenetic protein-2 and macrophage inhibitory cytokine-1 as mediators of the antiangiogenic activity of N-(4-hydroxyphenyl)retinamide. Clin Cancer Res 11(12):4610–4619. CrossRefPubMedGoogle Scholar
  14. 14.
    Samuel W, Kutty RK, Nagineni S, Vijayasarathy C, Chandraratna RA, Wiggert B (2006) N-(4-hydroxyphenyl)retinamide induces apoptosis in human retinal pigment epithelial cells: retinoic acid receptors regulate apoptosis, reactive oxygen species generation, and the expression of heme oxygenase-1 and Gadd153. J Cell Physiol 209(3):854–865. CrossRefPubMedGoogle Scholar
  15. 15.
    O'Brien J, Hayder H, Zayed Y, Peng C (2018) Overview of MicroRNA biogenesis, mechanisms of actions, and circulation. Front Endocrinol (Lausanne) 9:402. CrossRefGoogle Scholar
  16. 16.
    Zhuang G, Wu X, Jiang Z, Kasman I, Yao J, Guan Y, Oeh J, Modrusan Z, Bais C, Sampath D, Ferrara N (2012) Tumour-secreted miR-9 promotes endothelial cell migration and angiogenesis by activating the JAK-STAT pathway. EMBO J 31(17):3513–3523. CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Hu J, Zeng L, Huang J, Wang G, Lu H (2015) miR-126 promotes angiogenesis and attenuates inflammation after contusion spinal cord injury in rats. Brain Res 1608:191–202. CrossRefPubMedGoogle Scholar
  18. 18.
    Xu Z, Zhu C, Chen C, Zong Y, Feng H, Liu D, Feng W, Zhao J, Lu A (2018) CCL19 suppresses angiogenesis through promoting miR-206 and inhibiting Met/ERK/Elk-1/HIF-1alpha/VEGF-A pathway in colorectal cancer. Cell Death Dis 9(10):974. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Liu L, Bi N, Wu L, Ding X, Men Y, Zhou W, Li L, Zhang W, Shi S, Song Y, Wang L (2017) MicroRNA-29c functions as a tumor suppressor by targeting VEGFA in lung adenocarcinoma. Mol Cancer 16(1):50. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Isanejad A, Alizadeh AM, Amani Shalamzari S, Khodayari H, Khodayari S, Khori V, Khojastehnjad N (2016) MicroRNA-206, let-7a and microRNA-21 pathways involved in the anti-angiogenesis effects of the interval exercise training and hormone therapy in breast cancer. Life Sci 151:30–40. CrossRefPubMedGoogle Scholar
  21. 21.
    Umezu T, Imanishi S, Azuma K, Kobayashi C, Yoshizawa S, Ohyashiki K, Ohyashiki JH (2017) Replenishing exosomes from older bone marrow stromal cells with miR-340 inhibits myeloma-related angiogenesis. Blood Adv 1(13):812–823. CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Chakrabarti M, Khandkar M, Banik NL, Ray SK (2012) Alterations in expression of specific microRNAs by combination of 4-HPR and EGCG inhibited growth of human malignant neuroblastoma cells. Brain Res 1454:1–13. CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Herrero Martin D, Boro A, Schafer BW (2013) Cell-based small-molecule compound screen identifies fenretinide as potential therapeutic for translocation-positive rhabdomyosarcoma. PLoS ONE 8(1):e55072. CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Folkman J, Shing Y (1992) Angiogenesis. J Biol Chem 267(16):10931–10934PubMedGoogle Scholar
  25. 25.
    Otrock ZK, Makarem JA, Shamseddine AI (2007) Vascular endothelial growth factor family of ligands and receptors: review. Blood Cells Mol Dis 38(3):258–268. CrossRefPubMedGoogle Scholar
  26. 26.
    Golubkov V, Garcia A, Markland FS (2005) Action of fenretinide (4-HPR) on ovarian cancer and endothelial cells. Anticancer Res 25(1A):249–253PubMedGoogle Scholar
  27. 27.
    Bassani B, Bartolini D, Pagani A, Principi E, Zollo M, Noonan DM, Albini A, Bruno A (2016) Fenretinide (4-HPR) targets caspase-9, ERK 1/2 and the Wnt3a/beta-catenin pathway in medulloblastoma cells and medulloblastoma cell spheroids. PLoS ONE 11(7):e0154111. CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Tang NN, Zhu H, Zhang HJ, Zhang WF, Jin HL, Wang L, Wang P, He GJ, Hao B, Shi RH (2014) HIF-1alpha induces VE-cadherin expression and modulates vasculogenic mimicry in esophageal carcinoma cells. World J Gastroenterol 20(47):17894–17904. CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Yasuo M, Mizuno S, Allegood J, Kraskauskas D, Bogaard HJ, Spiegel S, Voelkel NF (2013) Fenretinide causes emphysema, which is prevented by sphingosine 1-phoshate. PLoS ONE 8(1):e53927. CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Liu X, Guan Y, Wang L, Niu Y (2017) MicroRNA-10b expression in node-negative breast cancer-correlation with metastasis and angiogenesis. Oncol Lett 14(5):5845–5852. CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Sheedy P, Medarova Z (2018) The fundamental role of miR-10b in metastatic cancer. Am J Cancer Res 8(9):1674–1688PubMedPubMedCentralGoogle Scholar
  32. 32.
    Plummer PN, Freeman R, Taft RJ, Vider J, Sax M, Umer BA, Gao D, Johns C, Mattick JS, Wilton SD, Ferro V, McMillan NA, Swarbrick A, Mittal V, Mellick AS (2013) MicroRNAs regulate tumor angiogenesis modulated by endothelial progenitor cells. Cancer Res 73(1):341–352. CrossRefPubMedGoogle Scholar
  33. 33.
    Zhu N, Zhang D, Xie H, Zhou Z, Chen H, Hu T, Bai Y, Shen Y, Yuan W, Jing Q, Qin Y (2011) Endothelial-specific intron-derived miR-126 is down-regulated in human breast cancer and targets both VEGFA and PIK3R2. Mol Cell Biochem 351(1–2):157–164. CrossRefPubMedGoogle Scholar
  34. 34.
    van Solingen C, Seghers L, Bijkerk R, Duijs JM, Roeten MK, van Oeveren-Rietdijk AM, Baelde HJ, Monge M, Vos JB, de Boer HC, Quax PH, Rabelink TJ, van Zonneveld AJ (2009) Antagomir-mediated silencing of endothelial cell specific microRNA-126 impairs ischemia-induced angiogenesis. J Cell Mol Med 13(8A):1577–1585. CrossRefPubMedGoogle Scholar
  35. 35.
    Wang S, Aurora AB, Johnson BA, Qi X, McAnally J, Hill JA, Richardson JA, Bassel-Duby R, Olson EN (2008) The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis. Dev Cell 15(2):261–271. CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Fish JE, Santoro MM, Morton SU, Yu S, Yeh RF, Wythe JD, Ivey KN, Bruneau BG, Stainier DY, Srivastava D (2008) miR-126 regulates angiogenic signaling and vascular integrity. Dev Cell 15(2):272–284. CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Lee S, Chen TT, Barber CL, Jordan MC, Murdock J, Desai S, Ferrara N, Nagy A, Roos KP, Iruela-Arispe ML (2007) Autocrine VEGF signaling is required for vascular homeostasis. Cell 130(4):691–703. CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Jing BQ, Ou Y, Zhao L, Xie Q, Zhang YX (2017) Experimental study on the prevention of liver cancer angiogenesis via miR-126. Eur Rev Med Pharmacol Sci 21(22):5096–5100. CrossRefPubMedGoogle Scholar
  39. 39.
    Chen H, Li L, Wang S, Lei Y, Ge Q, Lv N, Zhou X, Chen C (2014) Reduced miR-126 expression facilitates angiogenesis of gastric cancer through its regulation on VEGF-A. Oncotarget 5(23):11873–11885. CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Feng R, Chen X, Yu Y, Su L, Yu B, Li J, Cai Q, Yan M, Liu B, Zhu Z (2010) miR-126 functions as a tumour suppressor in human gastric cancer. Cancer Lett 298(1):50–63. CrossRefPubMedGoogle Scholar
  41. 41.
    Li XM, Wang AM, Zhang J, Yi H (2011) Down-regulation of miR-126 expression in colorectal cancer and its clinical significance. Med Oncol 28(4):1054–1057. CrossRefPubMedGoogle Scholar
  42. 42.
    Liu Y, Zhou Y, Feng X, An P, Quan X, Wang H, Ye S, Yu C, He Y, Luo H (2014) MicroRNA-126 functions as a tumor suppressor in colorectal cancer cells by targeting CXCR4 via the AKT and ERK1/2 signaling pathways. Int J Oncol 44(1):203–210. CrossRefPubMedGoogle Scholar
  43. 43.
    Zhang J, Du YY, Lin YF, Chen YT, Yang L, Wang HJ, Ma D (2008) The cell growth suppressor, mir-126, targets IRS-1. Biochem Biophys Res Commun 377(1):136–140. CrossRefPubMedGoogle Scholar
  44. 44.
    Saito Y, Friedman JM, Chihara Y, Egger G, Chuang JC, Liang G (2009) Epigenetic therapy upregulates the tumor suppressor microRNA-126 and its host gene EGFL7 in human cancer cells. Biochem Biophys Res Commun 379(3):726–731. CrossRefPubMedGoogle Scholar
  45. 45.
    Liu B, Peng XC, Zheng XL, Wang J, Qin YW (2009) MiR-126 restoration down-regulate VEGF and inhibit the growth of lung cancer cell lines in vitro and in vivo. Lung Cancer 66(2):169–175. CrossRefPubMedGoogle Scholar
  46. 46.
    Jusufovic E, Rijavec M, Keser D, Korosec P, Sodja E, Iljazovic E, Radojevic Z, Kosnik M (2012) let-7b and miR-126 are down-regulated in tumor tissue and correlate with microvessel density and survival outcomes in non–small–cell lung cancer. PLoS ONE 7(9):e45577. CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Li Z, Chen J (2011) In vitro functional study of miR-126 in leukemia. Methods Mol Biol 676:185–195. CrossRefPubMedGoogle Scholar
  48. 48.
    de Leeuw DC, Denkers F, Olthof MC, Rutten AP, Pouwels W, Schuurhuis GJ, Ossenkoppele GJ, Smit L (2014) Attenuation of microRNA-126 expression that drives CD34+38- stem/progenitor cells in acute myeloid leukemia leads to tumor eradication. Cancer Res 74(7):2094–2105. CrossRefPubMedGoogle Scholar
  49. 49.
    Sun Y, Bai Y, Zhang F, Wang Y, Guo Y, Guo L (2010) miR-126 inhibits non-small cell lung cancer cells proliferation by targeting EGFL7. Biochem Biophys Res Commun 391(3):1483–1489. CrossRefPubMedGoogle Scholar
  50. 50.
    Sun YQ, Zhang F, Bai YF, Guo LL (2010) miR-126 modulates the expression of epidermal growth factor-like domain 7 in human umbilical vein endothelial cells in vitro. Nan Fang Yi Ke Da Xue Xue Bao 30(4):767–770PubMedGoogle Scholar
  51. 51.
    Das E, Bhattacharyya NP (2014) MicroRNA-432 contributes to dopamine cocktail and retinoic acid induced differentiation of human neuroblastoma cells by targeting NESTIN and RCOR1 genes. FEBS Lett 588(9):1706–1714. CrossRefPubMedGoogle Scholar
  52. 52.
    Das S, Foley N, Bryan K, Watters KM, Bray I, Murphy DM, Buckley PG, Stallings RL (2010) MicroRNA mediates DNA demethylation events triggered by retinoic acid during neuroblastoma cell differentiation. Cancer Res 70(20):7874–7881. CrossRefPubMedPubMedCentralGoogle Scholar
  53. 53.
    Foley NH, Bray I, Watters KM, Das S, Bryan K, Bernas T, Prehn JH, Stallings RL (2011) MicroRNAs 10a and 10b are potent inducers of neuroblastoma cell differentiation through targeting of nuclear receptor corepressor 2. Cell Death Differ 18(7):1089–1098. CrossRefPubMedPubMedCentralGoogle Scholar
  54. 54.
    Poliakov E, Samuel W, Duncan T, Gutierrez DB, Mata NL, Redmond TM (2017) Inhibitory effects of fenretinide metabolites N-[4-methoxyphenyl]retinamide (MPR) and 4-oxo-N-(4-hydroxyphenyl)retinamide (3-keto-HPR) on fenretinide molecular targets beta-carotene oxygenase 1, stearoyl-CoA desaturase 1 and dihydroceramide Delta4-desaturase 1. PLoS ONE 12(4):e0176487. CrossRefPubMedPubMedCentralGoogle Scholar

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© Springer Nature B.V. 2020

Authors and Affiliations

  1. 1.Department of Medical Biology, Faculty of MedicineGaziantep UniversityGaziantepTurkey

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