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The anticancer activity of visnagin, isolated from Ammi visnaga L., against the human malignant melanoma cell lines, HT 144


Melanoma is a cancer of melanocyte cells and has the highest global incidence. There is a need to develop new drugs for the treatment of this deadly cancer, which is resistant to currently used treatment modalities. We investigated the anticancer activity of visnagin, a natural furanochromone derivative, isolated from Ammi visnaga L., against malignant melanoma (HT 144) cell lines. The singlet oxygen production capacity of visnagin was determined by the RNO bleaching method while cytotoxic activity by the MTT assay. Further, HT 144 cells treated with visnagin were also exposed to visible light (λ ≥ 400 nm) for 25 min to examine the illumination cytotoxic activity. The apoptosis was measured by flow cytometry with annexin V/PI dual staining technique. The effect of TNF-α secretion on apoptosis was also investigated. In standard MTT assay, visnagin (100 µg/mL) exhibited 80.93% inhibitory activity against HT 144 cancer cell lines, while in illuminated MTT assay at same concentration it showed lesser inhibitory activity (63.19%). Visnagin was induced apoptosis due to the intracellular generation of reactive oxygen species (ROS) and showed an apoptotic effect against HT 144 cell lines by 25.88%. However, it has no effect on TNF-α secretion. Our study indicates that visnagin can inhibit the proliferation of malignant melanoma, apparently by inducing the intracellular oxidative stress.

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  1. Bamhill RL, Fandrey K, Levy MA, Mihm ML, Hyman B (1992) Angiogenesis and tumor progression of melanoma. Quantification of vascularity in melanocytic nevi and cutaneous malign melanoma. Lab Invest 67:331–337

    Google Scholar 

  2. Watson M (2012) Drugs in clinical development for melanoma: summary and table. Pharm Med 26:171–183

    Article  Google Scholar 

  3. Rogers HW, Weinstock MA, Hinckley MR, Feldman SR, Fleischer AB, Coldiron BM (2010) Incidence estimate of nonmelanoma skin cancer in the United States, 2006. Arch Dermatol 146:283–287

    Article  PubMed  Google Scholar 

  4. Matsuo Y, Kamitani T (2010) Parkinson’s disease-related protein, a-synuclein, in malignant melanoma. PLoS ONE 5:1–8

    Google Scholar 

  5. Disse M, Reich H, Lee PK, Schram SS (2016) A review of the association between parkinson disease and malignant melanoma. Dermatol Surg 42:141–146

    Article  CAS  PubMed  Google Scholar 

  6. Thompson JF, Scolyer RA, Kefford RF (2005) Cutaneous melanoma. Lancet 365:687–701

    Article  CAS  PubMed  Google Scholar 

  7. Batistatou A, Cook MG, Massi D (2009) Histopathology report of cutaneous melanoma and sentinel lymph node in Europe: a web-based survey by the Dermatopathology Working Group of the European Society of Pathology. Virchows Arch 454:505–511

    Article  PubMed  Google Scholar 

  8. Soengas MS, Lowe SW (2003) Apoptosis and melanoma chemoresistance. Oncogene 22:3138–3151

    Article  CAS  PubMed  Google Scholar 

  9. Gunaydin K, Beyazit N (2004) The chemical investigations on the ripe fruits of Ammi visnaga (Lam.) Lamarck growing in Turkey. Nat Prod Res 18:169–175

    Article  CAS  PubMed  Google Scholar 

  10. Rauwald HW, Brehm O, Odenthal KP (1994) The Involvement of a Ca2+ channel blocking mode of action in the pharmacology of Ammi visnaga fruits. Planta Med 60:101–105

    Article  CAS  PubMed  Google Scholar 

  11. Martelli P, Bovalini L, Fe S, Franchi GG, Bari M (1985) Active oxygen forms in photoreaction between DNA and furanochromones khellin and visnagin. FEBS Lett 189:255–257

    Article  CAS  PubMed  Google Scholar 

  12. Chen X, Kagan J (1993) Photosensitized cleavage and cross-linking of pBR322 DNA with khellin and visnagin. J Photochem Photobiol B 20:183–189

    Article  CAS  PubMed  Google Scholar 

  13. Cuong TD, Lim CJ, Kim SW, Park JE, Hung TM, Min BS (2011) Isolation of compounds from Cimicifugae Rhizoma and their cytotoxic activity. Nat Prod Sci 17:80–84

    CAS  Google Scholar 

  14. El-Nakkady SS, Roaiah HF, El-Serwy WS, Soliman AM, El-Moez SIA, Abdel-Rahman AA-H (2012) Antitumor and antimicrobial activities of some hetero aromatic benzofurans derived from naturally occurring visnagin. Acta Pol Pharm 69:645–655

    CAS  PubMed  Google Scholar 

  15. Pakfetrat H, Nemati N, Shiravi A (2015) Cytotoxicity effects of Ammi visnaga extract on Hela and MCF-7 cancer cell line. Anim Biol 7:25–33

    Google Scholar 

  16. Kraljic I, Mohsni S (1978) A new method for the detection of singlet oxygen in aqueous solutions. Photochem Photobiol 28:577–581

    Article  CAS  Google Scholar 

  17. Aydoğmuş-Öztürk F, Günaydin K, Öztürk M, Jahan H, Duru ME, Choudhary MI (2018) Effect of Sideritis leptoclada against HT-144 human malignant melanoma. Melanoma Res 28:502–509

    Article  PubMed  Google Scholar 

  18. Wu D, Yotnda P (2011) Production and detection of reactive oxygen species (ROS) in cancer. J Vis Exp 57:1–4

    Google Scholar 

  19. Jahan H, Choudhary MI, Shah Z, Khan KM (2017) Derivatives of 6-nitrobenzimidazole inhibit fructose-mediated protein glycation and intracellular reactive oxygen species production. Med Chem 13:577–584

    Article  CAS  PubMed  Google Scholar 

  20. Vermes I, Haanen C, Steffensnakken H, Reutelingsperger C (1995) A novel assay for apoptosis-flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein-labeled annexin-V. J Immunol Methods 184:39–51

    Article  CAS  PubMed  Google Scholar 

  21. Iram N, Mildner M, Prior M, Petzelbauer P, Fiala C, Hacker S, Schoppl A, Tschachler E, Elbe-Burger A (2012) Age-related changes in expression and function of Toll-like receptors in human skin. Development 139:4210–4219

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bristow MR, Mason JW, Billingham ME, Daniels JR (1978) Doxorubicin cardiomyopathy: evaluation by phonocardiography, endomyocardial biopsy, and cardiac catheterization. Ann Intern Med 88:168–175

    Article  CAS  PubMed  Google Scholar 

  23. Xi L (2016) Visnagin—a new protectant against doxorubicin cardiotoxicity? Inhibition of mitochondrial malate dehydrogenase 2 (MDH2) and beyond. Ann Transl Med 4:65–69

    Article  PubMed  PubMed Central  Google Scholar 

  24. Sayed H, Mohamed MH, Farag SF, Mohamed GA, Proksch P (2007) A New Steroid Glycoside and Furochromones from Cyperus rotundus L. Nat Prod Res 21:343–350

    Article  CAS  PubMed  Google Scholar 

  25. Beltagy AM, Beltagy DM (2015) Chemical composition of Ammi visnaga L. and new cytotoxic activity of its constituents khellin and visnagin. J Pharm Sci Res 7:285–291

    CAS  Google Scholar 

  26. Circu ML, Aw TY (2010) Reactive oxygen species, cellular redox system and apoptosis. Free Radic Biol Med 48:749–762

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Matthews N, Neale ML, Jackson SK, Stark JM (1987) Tumour cell killing by tumour necrosis factor: inhibition by anaerobic conditions, free-radical scavengers and inhibitors of arachidonate metabolism. Immunology 62:153–155

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Larrick JW, Wright SC (1990) Cytotoxic mechanism of tumor necrosis factor-alpha. FASEB J 4:3215–3223

    Article  CAS  PubMed  Google Scholar 

  29. Shakibaei M, Schulze-Tanzil G, Takada Y, Aggarwal BB (2005) Redox regulation of apoptosis by members of the TNF superfamily. Antioxid Redox Signal 7:482–496

    Article  CAS  PubMed  Google Scholar 

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This study is a part of F.A.Ö’s Ph.D. thesis and was supported by the Research Fund of Istanbul University (Project Number: TP-19969). Prof. Dr. KerimanGünaydın would like to thank all staff of the Dr. Panjwani Center for Molecular Medicine and Drug Research (ICCBS), University of Karachi, Pakistan, for providing research facilities for her studies.

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Correspondence to Keriman Günaydın.

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Aydoğmuş-Öztürk, F., Jahan, H., Beyazit, N. et al. The anticancer activity of visnagin, isolated from Ammi visnaga L., against the human malignant melanoma cell lines, HT 144. Mol Biol Rep 46, 1709–1714 (2019).

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  • Visnagin
  • Ammi visnaga L.
  • Malignant melanoma
  • Apoptosis