Potential Cytotoxic Activity of Galegine on Human Melanoma Cells

Document Type : Original paper

Authors

1 Clinical Biochemistry Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.

2 Department of Medical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

3 Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, ‎Tehran, Iran.

4 Department of Medical Laboratory Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran.

5 Department of Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

6 Department of Physiology and Pharmacology, Faculty of Medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran.

Abstract

Background and objectives: Melanoma, the most lethal type of skin cancer, has a high recurrence rate within one year in melanoma patients following treatment by chemotherapy or immunotherapy. In an effort toward reducing this event, the present study aimed to investigate whether galegine has inhibitory effects on human melanoma cell lines. Galegine is a natural active compound found in Galega officinalis which was known and used in Europe for medicinal purposes for centuries.  Methods: Cell viability by MTT assay was conducted to measure the 50% inhibitory concentration (IC50) of galegine on DFW and SK-MEL-5 cells. Also, apoptosis level was determined using Annexin V/FITC-propidium iodide (PI) flow cytometry. In addition, quantitative real-time PCR (qRT-PCR) for Bax, Bcl2, and p53 genes was performed with specific primers to evaluate their expression pattern in each group. Results: The experimental results indicated that galegine induced cytotoxicity in a concentration-dependent manner with IC50 of 630 µM and 3300 µM in DFW and SK-MEL-5 cells, respectively. Also, apoptosis induction occurred in both melanoma cell lines, in a way that 12.4% of the DFW cells and 41.8% of SK-MEL-5 were detected in the apoptotic phase. Furthermore, it was found that the Bax/Bcl-2 ratio was upregulated in both melanoma cells. An upregulation in p53 gene expression was observed in SK-MEL-5 cells, as well. Conclusion: The results of the present study revealed that galegine induced cytotoxicity and apoptosis in human melanoma cells with the potential toward more research as a novel therapeutic candidate for melanoma treatment.

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Main Subjects

References

  • Patel H, Yacoub N, Mishra R, White A, Long Y, Alanazi S, Garrett JT. Current advances in the treatment of BRAF-mutant melanoma. Cancers(Basel). 2020; 12(2): 1–44.
  • Griffin M, Scotto D, Josephs DH, Mele S, Crescioli S, Bax HJ, Pellizzari G, Wynne MD, Nakamura M, Hoffmann RM, Ilieva KM, Cheung A, Spicer JF, Papa S, Lacy KE, Karagiannis SN. BRAF inhibitors: resistance and the promise of combination treatments for melanoma. Oncotarget. 2017; 8(44): 78174–78192.
  • Soengas MS, Lowe SW. Apoptosis and melanoma chemoresistance. Oncogene. 2003; 22(20): 3138–3151.
  • Miller AJ, Mihm MC. Melanoma. N Engl J Med. 2006; 355(1): 51–65.
  • American Cancer Society. 5-Year relative survival rates for melanoma skin cancer. [Accessed 2021]. Available from: https://www.cancer.org/cancer/melanoma-skin-cancer/detection-diagnosis-staging/survival-rates-for-melanoma-skin-cancer-by-stage.html.
  • Meier P, Finch A, Evan G. Apoptosis in development. Nature. 2000; 407(6805): 796–801.
  • Zhang YP, Li YQ, Lv YT, Wang JM. Effect of curcumin on the proliferation, apoptosis, migration, and invasion of human melanoma A375 cells. Genet Mol Res. 2015; 14(1): 1056–1067.
  • Wu Z, Liu B, Cailing E, Liu J, Zhang Q, Liu J, Chen N, Chen R, Zhu R. Resveratrol inhibits the proliferation of human melanoma cells by inducing G1/S cell cycle arrest and apoptosis. Mol Med Rep. 2015; 11(1): 400–404.
  • Mollazadeh H, Afshari AR, Hosseinzadeh H. Review on the potential therapeutic roles of Nigella sativa in the treatment of patients with cancer: involvement of apoptosis: black cumin and cancer. J Pharmacopuncture. 2017; 20(3): 158–172.
  • Sadeghnia HR, Jamshidi R, Afshari AR, Mollazadeh H, Forouzanfar F, Rakhshandeh H. Terminalia chebula attenuates quinolinate-induced oxidative PC12 and OLN-93 cell death. Mult Scler Relat Disord. 2017; 14: 60–67.
  • Afshari AR, Karimi Roshan M, Soukhtanloo M, Ghorbani A, Rahmani F, Jalili-Nik M, Vahedi MM, Hoseini A, Sadeghnia HR, Mollazadeh H, Mousavi SH. Cytotoxic effects of auraptene against a human malignant glioblastoma cell line. Avicenna J Phytomed. 2019; 9(4): 334–346.
  • Afshari AR, Jalili-Nik M, Soukhtanloo M, Ghorbani A, Sadeghnia HR, Mollazadeh H, Karimi Roshan M, Rahmani F, Sabri H, Vahedi MM, Mousavi SH. Auraptene-induced cytotoxicity mechanisms in human malignant glioblastoma (U87) cells: role of reactive oxygen species (ROS). EXCLI J. 2019; 18: 576–590.
  • Afshari AR, Roshan MK, Soukhtanloo M, Askari VR, Mollazadeh H, Nik MJ, Yazdi AAJ, Kia FA, Mousavi SH. Investigation of cytotoxic and apoptogenic effects of Terminalia chebula hydro-alcoholic extract on glioblastoma cell line. Shefaye Khatam. 2018; 6(4): 14–23.
  • Davoodi P, Ghoreishi SM, Hedayati A. Optimization of supercritical extraction of galegine from Galega officinalis: neural network modeling and experimental optimization via response surface methodology. Korean J Chem Eng. 2016; 34(3): 854–865.
  • Witters LA. The blooming of the French lilac. J Clin Invest. 2001; 108(8): 1105–1107.
  • Penagos Tabares F, Bedoya Jaramillo JV, Ruiz-Cortés ZT. Pharmacological overview of galactogogues. Vet Med Int. 2014; Article ID 602894.
  • Mooney MH, Fogarty S, Stevenson C, Gallagher AM, Palit P, Hawley SA, Hardie DG, Coxon GD, Waigh RD, Tate RJ, Harvey AL, Furman BL. Mechanisms underlying the metabolic actions of galegine that contribute to weight loss in mice. Br J Pharmacol. 2008; 153(8): 1669–1677.
  • Park JH, Kim YH, Park EH, Lee SJ, Kim H, Kim A, Lee SB, Shim S, Jang H, Myung JK, Park S, Lee SJ, Kim MJ. Effects of metformin and phenformin on apoptosis and epithelial-mesenchymal transition in chemoresistant rectal cancer. Cancer Sci. 2019; 110(9): 2834–2845.
  • Huxtable CR, Dorling PR, Colegate SM. Identification of galegine, an isoprenyl guanidine, as the toxic principle of Schoenus asperocarpus (poison sedge). Aust Vet J. 1993; 70(5): 169–171.
  • Lee JS, Kim WS, Kim JJ, Chin YW, Jeong HC, Choi JS, Min HG, Cha HJ. Identification of anti-melanogenic natural compounds from Galega officinalis and further drug repositioning. J Dermatol Sci. 2012; 67(1): 61–63.
  • Safe S, Nair V, Karki K. Metformin-induced anticancer activities: recent insights. Biol Chem. 2018; 399(4): 321–335.
  • Garcia Rubino ME, Carrillo E, Ruiz Alcala G, Dominguez-Martin A, Juan AM, Boulaiz H. Phenformin as an anticancer agent: challenges and prospects. Int J Mol Sci. 2019; 20(13): 1–17.
  • Bridges HR, Jones AJ, Pollak MN, Hirst J. Effects of metformin and other biguanides on oxidative phosphorylation in mitochondria. Biochem J. 2014; 462(3): 475–487.
  • Geoghegan F, Chadderton N, Farrar GJ, Zisterer DM, Porter RK. Direct effects of phenformin on metabolism/bioenergetics and viability of SH-SY5Y neuroblastoma cells. Oncol Lett. 2017; 14(5): 6298–6306.
  • Ali Z, Yousaf N, Larkin J. Melanoma epidemiology, biology and prognosis. EJC Suppl. 2013; 11(2): 81–91.
  • Schadendorf D, van Akkooi ACJ, Berking C, Griewank KG, Gutzmer R, Hauschild A, Stang A, Roesch A, Ugurel S. Melanoma. Lancet. 2018; 392(10151): 971–984.
  • Pasquali S, Hadjinicolaou AV, Chiarion Sileni V, Rossi CR, Mocellin S. Systemic treatments for metastatic cutaneous melanoma. Cochrane Database Syst Rev. 2018; Article ID CD011123.
  • Yang TY, Wu YJ, Chang CI, Chiu CC, Wu ML. The effect of bornyl cis-4-hydroxycinnamate on melanoma cell apoptosis is associated with mitochondrial dysfunction and endoplasmic reticulum stress. Int J Mol Sci. 2018; 19(5): 1–18.
  • Sharma A, Kaur M, Katnoria J, Nagpal A. Polyphenols in food: cancer prevention and apoptosis induction. Curr Med Chem. 2018; 25(36): 4740–4757.
  • Mousavi SH, Tayarani-Najaran Z, Hersey P. Apoptosis: from signalling pathways to therapeutic tools. Iran J Basic Med Sci. 2008; 11(3): 121–142.
  • Parsaee H, Asili J, Mousavi SH, Soofi H, Emami SA, Tayarani-Najaran Z. Apoptosis induction of Salvia chorassanica root extract on human cervical cancer cell line. Iran J Pharm Res. 2013; 12(1): 75–83.
  • Piperigkou Z, Manou D, Karamanou K, Theocharis AD. Strategies to target matrix metalloproteinases as therapeutic approach in cancer. Methods Mol Biol. 2018; 1731: 325–348.
  • Shi L, Chen J, Yang J, Pan T, Zhang S, Wang Z. MiR-21 protected human glioblastoma U87MG cells from chemotherapeutic drug temozolomide induced apoptosis by decreasing Bax/Bcl-2 ratio and caspase-3 activity. Brain Res. 2010; 1352: 255–264.
  • Jiang Z, Zheng X, Rich KM. Down-regulation of Bcl-2 and Bcl-xL expression with bispecific antisense treatment in glioblastoma cell lines induce cell death. J Neurochem. 2003; 84(2): 273–281.
  • Bode AM, Dong Z. Post-translational modification of p53 in tumorigenesis. Nat Rev Cancer. 2004; 4(10): 793–805.
  • Tapia N, Scholer HR. P53 connects tumorigenesis and reprogramming to pluripotency. J Exp Med. 2010; 207(10): 2045–2048.
  • Clewell RA, Sun B, Adeleye Y, Carmichael P, Efremenko A, McMullen PD, Pendse S, Trask OJ, White A, Andersen ME. Profiling dose-dependent activation of p53-mediated signaling pathways by chemicals with distinct mechanisms of DNA damage. Toxicol Sci. 2014; 142(1): 56–73.

 

Research Journal of Pharmacognosy
Volume 9, Issue 3
July 2022
Pages 15-23

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