Exploring the Antiproliferative Mechanisms of Urtica dioica L. extract in Human Promyelocytic Leukemia Cell Line

Yıl 2021, , 468 - 474, 29.12.2021

Ebru Temiz , İsmail Koyuncu , Shameem Saadat , Özgür Yüksekdağ Yazan Award

https://doi.org/10.35440/hutfd.1012538

Cited By: 3

Öz

Background: Nature is a source of therapeutic compounds which have fewer side effects compared to synthetic drugs. Stinging nettle extract, widely-used in Anatolia, has a strong antiproliferative effect on many types of cancer. However, the underlying molecular mechanisms of this effect is still not known.
Materials and Methods: In this study, the interaction of Urtica dioica L. extract at different concentrations with apoptosis and autophagy pathways in the human promyelocytic cell line (HL-60) was studied to elucidate how it triggers the antiproliferative effects. In this context, firstly, the plant leaves were extracted in water with the Soxhlet extraction method. HL-60 cells were incubated with the extract at different concentrations for 24 hours, and the activated antitumoral effect pathway was investigated with advanced following molecular tests: MTT staining, Nitric oxide (NO) level, Annexin-V in flow cytometry, cell cycle, mitochondrial membrane potential measurement (MMP) and qPCR for evaluation of apoptosis and autophagy mediator genes.
Results: It was determined that cell proliferation was suppressed at a concentration of 100 µg/mL and cells were kept in G0/G1 phase, MMP was impaired in cells and the rate of apoptotic cells increased. These apoptotic markers were confirmed by statistically increased expression of apoptotic and autophagy genes and NO level.
Conclusions: Taken together, it is predicted that Urtica dioica L. water extract initiates apoptosis in HL-60 cells and could be promising compound candidate for cancer treatment.

Anahtar Kelimeler

HL-60, Urtica dioica, Cancer, Apoptosis, APL

İnsan Promiyelositik Lösemi Hücre Hattında Isırgan Ekstraktının Antiproliferatif Mekanizmalarının Araştırılması

Öz

Amaç: Doğa, sentetik ilaçlara kıyasla daha az yan etkiye sahip terapötik bileşiklerin kaynağıdır. Anadolu'da yaygın olarak kullanılan ısırgan otu özütü, birçok kanser türü üzerinde güçlü bir antiproliferatif etkiye sahiptir. Bununla birlikte, bu etkinin altında yatan moleküler mekanizmalar hala bilinmemektedir.
Materyal ve Metod: Bu çalışmada, insan promiyelositik hücre hattında (HL-60) farklı konsantrasyonlarda hazırlanan ısırgan ekstraktının apoptozis ve otofaji yolakları ile etkileşimi ve antiproliferatif etkileri incelenmiştir. Bu kapsamda öncelikle bitki yaprakları Soxhlet ekstraksiyon yöntemi ile suda ekstrakte edilmiştir. HL-60 hücreleri, 24 saat boyunca farklı konsantrasyonlarda ekstrakt ile inkübe edildi ve aktif anti-tümöral etki yolu, ileri moleküler testler ile araştırıldı: MTT boyama, Nitrik oksit (NO) seviyesi, flow sitometride Annexin-V, hücre döngüsü, mitokondriyal zar potansiyeli ölçümü (MMP) ile apoptozis ve otofaji yolak genlerinin değerlendirilmesi için qPCR.
Bulgular: 100 µg/mL konsantrasyonda hücre proliferasyonunun baskılandığı ve hücrelerin G0/G1 fazında tutulduğu, hücrelerde MMP'nin bozulduğu ve apoptotik hücre yüzdesinin arttığı belirlendi. Bu apoptotik belirteçler, apoptotik ve otofaji genlerinin istatistiksel olarak artan ekspresyonu ve NO seviyesi ile doğrulandı.
Sonuç: Tüm sonuçlar birlikte ele alındığında, ısırgan su ekstraktının HL-60 hücrelerinde apoptozisi başlattığı ve kanser tedavisi için umut verici bitki adayı olabileceği tahmin edilmektedir.

Anahtar Kelimeler

HL-60, Isırgan, Kanser, Apoptozis, APL

Kaynakça

• 1. Jalili M, Salehzadeh-Yazdi A, Mohammadi S, Yaghmaie M, Ghavamzadeh A, and Alimoghaddam K. Meta-Analysis of Gene Expression Profiles in Acute Promyelocytic Leukemia Reveals Involved Pathways. Int J Hematol Oncol Stem Cell Res. 2017; 11(1): 1-12.
• 2. Ohno R, Asou N, and Ohnishi K. Treatment of acute promyelocytic leukemia: strategy toward further increase of cure rate. Leukemia. 2003; 17(8): 1454-63.
• 3. Lo-Coco F and Hasan SK. Understanding the molecular pathogenesis of acute promyelocytic leukemia. Best Pract Res Clin Haematol. 2014; 27(1): 3-9.
• 4. Mohammadi A, Mansoori B, Aghapour M, Shirjang S, Nami S, and Baradaran B. The Urtica dioica extract enhances sensitivity of paclitaxel drug to MDA-MB-468 breast cancer cells. Biomed Pharmacother. 2016; 83: 835-42.
• 5. Mohammadi A, Mansoori B, Aghapour M, and Baradaran B. Urtica dioica dichloromethane extract induce apoptosis from intrinsic pathway on human prostate cancer cells (PC3). Cell Mol Biol (Noisy-le-grand). 2016; 62(3): 78-83.
• 6. Mohammadi A, Mansoori B, Goldar S, Shanehbandi D, Khaze V, Mohammadnejad L, et al. Effects of Urtica dioica dichloromethane extract on cell apoptosis and related gene expression in human breast cancer cell line (MDA-MB-468). Cell Mol Biol (Noisy-le-grand). 2016; 62(2): 62-7.
• 7. Bnouham M, Merhfour FZ, Ziyyat A, Aziz M, Legssyer A, and Mekhfi H. Antidiabetic effect of some medicinal plants of Oriental Morocco in neonatal non-insulin-dependent diabetes mellitus rats. Hum Exp Toxicol. 2010; 29(10): 865-71.
• 8. El Haouari M, Bnouham M, Bendahou M, Aziz M, Ziyyat A, Legssyer A, et al. Inhibition of rat platelet aggregation by Urtica dioica leaves extracts. Phytother Res. 2006; 20(7): 568-72.
• 9. Konrad L, Muller HH, Lenz C, Laubinger H, Aumuller G, and Lichius JJ. Antiproliferative effect on human prostate cancer cells by a stinging nettle root (Urtica dioica) extract. Planta Med. 2000; 66(1): 44-7.
• 10. Testai L, Chericoni S, Calderone V, Nencioni G, Nieri P, Morelli I, et al. Cardiovascular effects of Urtica dioica L. (Urticaceae) roots extracts: in vitro and in vivo pharmacological studies. J Ethnopharmacol. 2002; 81(1): 105-9.
• 11. Mansoori B, Mohammadi A, Shir Jang S, and Baradaran B. Mechanisms of immune system activation in mammalians by small interfering RNA (siRNA). Artif Cells Nanomed Biotechnol. 2016; 44(7): 1589-96.
• 12. Mansoori B, Sandoghchian Shotorbani S, and Baradaran B. RNA interference and its role in cancer therapy. Adv Pharm Bull. 2014; 4(4): 313-21.
• 13. Roy P and Saikia B. Cancer and cure: a critical analysis. Indian journal of cancer. 2016; 53(3): 441.
• 14. Esposito S, Bianco A, Russo R, Di Maro A, Isernia C, and Pedone PV. Therapeutic perspectives of molecules from Urtica dioica extracts for cancer treatment. Molecules. 2019; 24(15): 2753.
• 15. Demiryürek AT, Saracaloglu A, Temiz E, Demiryürek S, and Sener B. Investigation of the Apoptotic Effects of Pistacia vera Nut Skin Methanol Extract on Human Gastric Cancer Cell Line HGC-27. in Multidisciplinary Digital Publishing Institute Proceedings. 2018.
• 16. Yumrutaş Ö, Pehlivan M, Güven C, Bozgeyik I, Bozgeyik E, YUMRUTAŞ P, et al. Investigation of cytotoxic effect of salvia pilifera extracts and synthetic chlorogenic and caffeic acids on DU145 prostate cancer cells line. Tarim ve Doga Dergisi. 2018; 21(2): 141.
• 17. Nobili S, Lippi D, Witort E, Donnini M, Bausi L, Mini E, et al. Natural compounds for cancer treatment and prevention. Pharmacological research. 2009; 59(6): 365-78.
• 18. Mohammadi A, Mansoori B, Aghapour M, Shirjang S, Nami S, and Baradaran B. The Urtica dioica extract enhances sensitivity of paclitaxel drug to MDA-MB-468 breast cancer cells. Biomedicine & Pharmacotherapy. 2016; 83: 835-42.
• 19. Rejhová A, Opattová A, Čumová A, Slíva D, and Vodička P. Natural compounds and combination therapy in colorectal cancer treatment. European journal of medicinal chemistry. 2018; 144: 582-94.
• 20. Nahata A, Saxena A, Suri N, Saxena AK, and Dixit VK. Sphaeranthus indicus induces apoptosis through mitochondrial-dependent pathway in HL-60 cells and exerts cytotoxic potential on several human cancer cell lines. Integrative cancer therapies. 2013; 12(3): 236-47.
• 21. Turk S, Malkan UY, Ghasemi M, Hocaoglu H, Mutlu D, Gunes G, et al. Growth inhibitory activity of Ankaferd hemostat on primary melanoma cells and cell lines. SAGE open medicine. 2017; 5: 205.
• 22. Kardan M, Rafiei A, Golpour M, Ebrahimzadeh MA, Akhavan-Niaki H, and Fattahi S. Urtica dioica Extract Inhibits Cell Proliferation and Induces Apoptosis in HepG2 and HTC116 as Gastrointestinal Cancer Cell Lines. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents). 2020; 20(8): 963-69.
• 23. Bock FJ and Tait SW. Mitochondria as multifaceted regulators of cell death. Nature reviews Molecular cell biology. 2020; 21(2): 85-100.
• 24. Chen C-Y, Chen J, He L, and Stiles BL. PTEN: tumor suppressor and metabolic regulator. Frontiers in endocrinology. 2018; 9: 338.
• 25. Mercogliano MF, Bruni S, Elizalde PV, and Schillaci R. Tumor necrosis factor α blockade: an opportunity to tackle breast Cancer. Frontiers in oncology. 2020; 10(1):12-17.
• 26. Abbastabar M, Kheyrollah M, Azizian K, Bagherlou N, Tehrani SS, Maniati M, et al. Multiple functions of p27 in cell cycle, apoptosis, epigenetic modification and transcriptional regulation for the control of cell growth: a double-edged sword protein. DNA repair. 2018; 69: 63-72.
• 27. Marquez RT and Xu L. Bcl-2: Beclin 1 complex: multiple, mechanisms regulating autophagy/apoptosis toggle switch. American journal of cancer research. 2012; 2(2): 214.
• 28. Bostancıklıoğlu M. An update on the interactions between Alzheimer's disease, autophagy and inflammation. Gene. 2019; 705: 157-66.
• 29. Mutlu H, Mutlu S, and Bostancıklıoğlu M. Profiling of Autophagy-Associated microRNAs in the Osteosarcoma Cell Line of U2OS. Anti-Cancer Agents in Medicinal Chemistry. 2020; 2021(21): 1-7.
• 30. Liu C, Xu P, Chen D, Fan X, Xu Y, Li M, et al. Roles of autophagy‑related genes Beclin‑1 and LC3 in the development and progression of prostate cancer and benign prostatic hyperplasia. Biomedical reports. 2013; 1(6): 855-60.
• 31. Choudhari SK, Chaudhary M, Bagde S, Gadbail AR, and Joshi V. Nitric oxide and cancer: a review. World journal of surgical oncology. 2013; 11(1): 1-11.
• 32. Olson SY and Garbán HJ. Regulation of apoptosis-related genes by nitric oxide in cancer. Nitric Oxide. 2008; 19(2): 170-76.
• 33. Harada K, Kawaguchi S-I, Tomitaro O, Yoshida H, and Sato M. Overexpression of iNOS gene suppresses the tumorigenicity and metastasis of oral cancer cells. in vivo. 2004; 18(4): 449-55.
• 34. Weiming X, Liu LZ, Loizidou M, Ahmed M, and Charles IG. The role of nitric oxide in cancer. Cell research. 2002; 12(5): 311-20.
• 35. Kim HH and Kim K. Enhancement of TNF‐α‐mediated cell death in vascular smooth muscle cells through cytochrome c‐independent pathway by the proteasome inhibitor. FEBS letters. 2003; 535(1-3): 190-94.