A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells

Baran, Yusuf; YAŞAR, Mustafa; GÖKBULUT, Aysun Adan

A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells

Aysun Adan GÖKBULUT, (İzmir Teknoloji Enstitüsü Moleküler Biyoloji ve Genetik Bölümü, İzmir, Türkiye)

Mustafa YAŞAR, (Naturin Doğal Ürünler, İzmir, Türkiye)

Yusuf BARAN (Abdullah Gül Üniversitesi, Tıp Fakültesi, Hayat ve Doğa Bilimleri, Bölümü, Kayseri, Türkiye)

Turkish Journal of Hematology

2 0

Yıl: 2015 Cilt: 32 Sayı: 2 Sayfa Aralığı: 118 - 126 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells

Öz:

Amaç: Bu çalışmanın amacı yeni bitkisel bir ürün olan KL-21in [naturin doğal ürünler şirketi (İzmir, Türkiye) tarafındanüretilen] 232B4 kronik lenfostik lösemi (KLL) hücreleri üzerindeki sitotoksik ve apoptotik etkilerinin araştırılmasıdır. Ayrıca,KL-21in BEAS-2B sağlıklı insan bronşial epitelyum hücreleri üzerindeki sitotoksik etkisine de bakılmıştır. Gereç ve Yöntemler: KL-21in sitotoksik etkisine MTT hücre çoğalma testiyle bakılmıştır. Kaspaz-3 enzim aktivitesindekive mitokondri membran potansiyelindeki değişimlere sırasıyla kaspaz-3 kolorimetrik testi ve JC-1 boyasına dayalı bir yöntemkullanılarak bakılmıştır. Apoptotik hücre popülasyonunu belirlemek amacıyla Anneksin 5-FITC/PI ikili boyama yöntemikullanılmıştır. KL-21in KLL hücrelerinin hücre siklusu üzerindeki etkilerine akım sitometresi ile bakılmıştır. Bulgular: KL-21in KLL hücrelerinin çoğalması üzerine etkisi zamana ve doza bağımlı olarak artmıştır. Bununlar beraber, KL- 21in özellikle yüksek konsantrasyonlarda KLL hücreleri ile karşılaştırıldığında BEAS-2B hücreleri üzerinde daha az sitotoksik etkigösterdiği saptanmıştır. Anneksin-5/PI ikili boyaması 232B4 hücrelerinde apoptotik hücre popülasyonunun arttığını göstermiştir.KL-21in artan konsantrayonları kaspaz-3 enzim aktivitesini arttırmış ve mitokondri membran potansiyelindeki kayıplarıindüklemiştir. KL-21 1 mg/ml konsantrasyonuna kadar G0/G1 fazındaki hücrelerin yüzdesinde küçük artışlara ve S fazındakihücre popülasyonunda ise azalmalara neden olmaktadır. En yüksek konsantrasyonda ise hücrelerin büyük bir çoğunluğu G0/G1fazında birikmiştir. Sonuç: Elde edilen sonuçlar, KL-21in 232B4 KLL hücreleri üzerinde büyümeyi inhibe edici bir etkisi olduğunu göstermiştir.Ayrıca, K-21 apoptozu indüklemekte ve G0/G1 fazında hücre siklusunun tutulumuna neden olmaktadır.

Anahtar Kelime:

Konular: Hematoloji

Yeni Doğal Bir Ürün Olan KL-21 Kronik Lenfositik Lösemi Hücrelerinin Çoğalmasını İnhibe Etmekte ve Apoptozu İndüklemektedir

Öz:

Objective: The aims of this study were to examine the cytotoxic and apoptotic effects of KL-21, a novel plant product(produced by naturin natural Products, İzmir, Turkey), on 232B4 chronic lymphocytic leukemia (CLL) cells and to determinethe cytotoxic effects on healthy BEAS-2B human bronchial epithelial cells.Materials and Methods: The cytotoxic effect of KL-21 was determined by MTT cell proliferation assay. Changes incaspase-3 enzyme activity were measured using the caspase-3 colorimetric assay. Changes in mitochondrial membranepotential were determined using the J C-1 dye-based method. Annexin V-FITC/PI double staining was performed to measurethe apoptotic cell population. Effects of KL-21 on cell cycle profiles of CLL cells were investigated by flow cytometry.Results: We detected time- and concentration-dependent increases in the cytotoxic effect of KL-21 on 232B4 CLL cells.However, we also showed that, especially at higher concentrations, KL-21 was less cytotoxic towards BEAS-2B healthy cellsthan towards CLL cells. Annexin-V/PI double staining results showed that the apoptotic cell population increased in 232B4cells. Increasing concentrations of KL-21 increased caspase-3 enzyme activity and induced loss of mitochondrial membranepotential. KL-21 administration resulted in small increases in the percentage of the cells in the G0/G1 phase while it decreasedthe S phase cell population up to 1 mg/mL. At the highest concentration, most of the cells accumulated in the G0/G1 phase.Conclusion:KL-21 has a growth-inhibitory effect on 232B4 CLL cells. KL-21 causes apoptosis and cell cycle arrest at G0/G1.

Anahtar Kelime:

Konular: Hematoloji

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Chiorazzi N, Ferrarini M. B cell chronic lymphocytic leukemia: lessons learned from studies of the B cell antigen receptor. Annu Rev Immunol 2003;21:841-894.
2. Caligaris-Cappio F, Ghia P. Novel insights in chronic lymphocytic leukemia: are we getting closer to understanding the pathogenesis of the disease? J Clin Oncol 2008;26:4497- 4503.
3. Rozman C, Montserrat E. Chronic lymphocytic leukemia. N Engl J Med 1995;333:1052-1057.
4. Veliz M, Pinilla-Ibarz J. Role of ofatumumab in treatment of chronic lymphocytic leukemia. J Blood Med 2011;2:71-77.
5. Byrd J C, Stilgenbauer S, Flinn IW. Chronic lymphocytic leukemia. Hematology 2004;2004:163-183.
6. Flinn IW, Grever MR. Chronic lymphocytic leukemia. Cancer Treat Rev 1996;96:1-13.
7. Hainsworth JD, Litchy S, Barton JH, Houston GA, Hermann RC, Bradof JE, Greco FA; Minnie Pearl Cancer Research Network. Single-agent rituximab as first-line and maintenance treatment for patients with chronic lymphocytic leukemia or small lymphocytic lymphoma: a phase II trial of the Minnie Pearl Cancer Research Network. J Clin Oncol 2003;21:1746- 1751.
8. OBrien SM, Kantarjian HM, Thomas DA, Cortes J, Giles FJ, Wierda WG, Koller CA, Ferrajoli A, Browning M, Lerner S, Albitar M, Keating MJ. Alemtuzumab as treatment for residual disease after chemotherapy in patients with chronic lymphocytic leukemia. Cancer 2003;98:2657-2663.
9. Dreger P, Montserrat E. Autologous and allogeneic stem cell transplantation for chronic lymphocytic leukemia. Leukemia 2002;16:985-992.
10. Esteve J, Villamor N, Colomer D, Cervantes F, Campo E, Carreras E, Montserrat E. Stem cell transplantation for chronic lymphocytic leukemia: different outcome after autologous and allogeneic transplantation and correlation with minimal residual disease status. Leukemia 2001;5:445-451.
11. Berdeja J G, Jones RJ, Zahurak ML, Piantadosi S, Abrams RA, Borowitz MJ, Vogelsang GB, Noga SJ, Ambinder RF, Flinn IW. Allogeneic bone marrow transplantation in patients with sensitive low-grade lymphoma or mantle cell lymphoma. Biol Blood Marrow Transplant 2001;7:561-567.
12. Veliz M, Pinilla-Ibarz J. Treatment of relapsed or refractory chronic lymphocytic leukemia. Cancer Control 2012;19:37- 53.
13. Podhorecka M, Halicka D, Klimek P, Kowal M, Chocholska S, Dmoszynska A. Resveratrol increases rate of apoptosis caused by purine analogues in malignant lymphocytes of chronic lymphocytic leukemia. Ann Hematol 2011;90:173-183.
14. Russo M, Spagnuolo C, Volpe S, Mupo A, Tedesco I, Russo GL. Quercetin induced apoptosis in association with death receptors and fludarabine in cells isolated from chronic lymphocytic leukaemia patients. Br J Cancer 2010;103:642- 648.
15. Baran Y, Bielawski J, Gunduz U, Ogretmen B. Targeting glucosylceramide synthase sensitizes imatinib-resistant chronic myeloid leukemia cells via endogenous ceramide accumulation. J Cancer Res Clin Oncol 2011;137:1535-1544.
16. Avcı ÇB, Gündüz C, Baran Y, Şahin F, Yılmaz S, Dogan ZO, Saydam G. Caffeic acid phenethyl ester triggers apoptosis through induction of loss of mitochondrial membrane potential in CCRF-CEM cells. J Cancer Res Clin Oncol 2011;137:41-47.
17. Brown JR. The treatment of relapsed refractory chronic lymphocytic leukemia. Hematology Am Soc Hematol Educ Program 2011;2011:110-118.
18. Battle TE, Arbiser J, Frank DA. The natural product honokiol induces caspase-dependent apoptosis in B-cell chronic lymphocytic leukemia (B-CLL) cells. Blood 2005;106:690- 697.
19. Gokbulut AA, Apohan E, Baran Y. Resveratrol and quercetin- induced apoptosis of human 232B4 chronic lymphocytic leukemia cells by activation of caspase-3 and cell cycle arrest. Hematology 2013;18:144-150.
20. König A, Schwartz GK, Mohammad RM, Al-Katib R, Gabrilove JL. The novel cyclin-dependent kinase inhibitor flavopiridol downregulates Bcl-2 and induces growth arrest and apoptosis in chronic B-cell leukemia lines. Blood 1997;90:4307-4312.
21. Csupor-Löffler B, Hajdú Z, Zupkó I, Réthy B, Falkay G, Forgo P, Hohmann J. Antiproliferative effect of flavonoids and sesquiterpenoids from Achillea millef olium s.l. on cultured human tumour cell lines. Phytother Res 2009;23:672-676.
22. Akbay P, Basaran AA , Undeger U, Basaran N. In vitro immunomodulatory activity of flavonoid glycosides from Urtica dioica L. Phytother Res 2003;17:34-37.
23. Yoshinobu Y. Antitumor activity of crude protein extracted from Equisetum arvense LINNE. Journal of Analytical Bio- Science 1992;22:421-424.
24. Asgarpanah J, Roohi E. Phytochemistry and pharmacological properties of Equisetum arvense L. J Med Plants Res 2012;6:3689-3693.
25. Jimenez-Arellanes A, Martinez R, García R, Leon-Diaz R, Luna-Herrera J, Molina-Salinas G, Said-Fernández S. Thymus vulgaris as a potencial source of antituberculous compounds. Pharmacologyonline 2006;3:569-574.
26. Kienle G. Influence of Viscum album L (European mistletoe) extracts on quality of life in cancer patients: a systematic review of controlled clinical studies. Integr Cancer Ther 2010;9:142-157.
27. Sabova L, Pilátová M, Szilagyi K, Sabo R, Mojzis J. Cytotoxic effect of mistletoe (Viscum album L.) extract on Jurkat cells and its interaction with doxorubicin. Phytother Res 2010;24:365- 368.
28. Singh R, Sharma PK, Malviya R. Pharmacological properties and ayurvedic value of Indian buch plant (Acorus calamus): a short review. Advances in Biological Research 2011;5:145-154.
29. Demir H, Açık L, Bali EB, Koç LY, Kaynak G. Antioxidant and antimicrobial activities of Solidago virgaurea extracts. Afr J Biotechnol 2009;8:274-279.
30. Gross SC, Goodarzi G, Watabe M, Bandyopadhyay S, Pai SK, Watabe K. Antineoplastic activity of Solidago virgaurea on prostatic tumor cells in an SCID mouse model. Nutr Cancer 2002;43:76-81.
31. Davis-Searles PR, Nakanishi Y, Kim NC, Graf TN, Oberlies NH, Wani MC, Wall ME, Agarwal R, Kroll DJ. Milk thistle and prostate cancer: differential effects of pure flavonolignans from Silybum marianum on antiproliferative end points in human prostate carcinoma cells. Cancer Res 2005;65:4448- 4457.
32. Huseini HF, Larijani B, Heshmat R, Fakhrzadeh H, Radjabipour B, Toliat T, Raza M. The efficacy of Silybum marianum (L.) Gaertn. (silymarin) in the treatment of type II diabetes: a randomized, double-blind, placebo-controlled, clinical trial. Phytother Res 2006;20:1036-1039.
33. Chattopadhyay I, Biswas K, Bandyopadhyay U, Banerjee RK. Turmeric and curcumin: biological actions and medicinal applications. Curr Sci India 2004;87:44-53.
34. Al-Sereitia MR, Abu-Amerb KM, Sena P. Pharmacology of rosemary (Rosmarinus off icinalis Linn.) and its therapeutic potentials. Indian J Exp Biol 1999;37:124-131.
35. Gören AC, Topçu G, Bilsel G, Bilsel M, Aydoğmus Z, Pezzuto JM. The chemical constituents and biological activity of essential oil of Lavandula stoechas ssp. stoechas. Z Naturforsch 2002;57c:797-800.
36. Neves JM, Matos C, Moutinho C, Queiroz E, Gomes LR. Ethnopharmacological notes about ancient uses of medicinal plants in Tras-os-Montes (northern of Portugal). J Ethnopharmacol 2009;124:270-283.
37. Manske RHF. The alkaloids of fumariaceous plants: xviii. Fumaria off icinalis L. Can J Res 1938;16:438-444.
38. Sigstedt SC, Hooten CJ, Callewaert MC, Jenkins AR, Romero AE, Pullin MJ, Kornienko A, Lowrey TK, Slambrouck SV, Steelant WF. Evaluation of aqueous extracts of Taraxacum off icinale on growth and invasion of breast and prostate cancer cells. Int J Oncol 2008;32:1085-1090.
39. Hu C, Kitts DD. Antioxidant, prooxidant, and cytotoxic activities of solvent-fractionated dandelion (Taraxacum off icinale) flower extracts in vitro. J. Agric Food Chem 2003;51:301-310.
40. Chen CJ, Deng AJ, Liu C, Shi R, Qin HL, Wang AP. Hepatoprotective activity of Cichorium endivia L. extract and its chemical constituents. Molecules 2011;16:9049-9066.
41. Al-Amin ZM, Thomson M, Al-Qattan KK, Peltonen-Shalaby R, Ali M. Anti-diabetic and hypolipidaemic properties of ginger (Zingiber off icinale) in streptozotocin-induced diabetic rats. Br J Nutr 2006;96:660-666.
42. Mirzaei M. Treatment of natural tropical theileriosis with the extract of the plant Peganum harmala. Korean J Parasitol 2007;45:267-271.
43. Lamchouri F, Settaf A, Cherrah Y, Zemzami M, Lyoussi B, Zaid A, Atif N, Hassar M. Antitumour principles from Peganum harmala seeds. Therapie 1999;54:753-758.
44. Pepeljnjak S, Kosalec I, Kalodera Z, Blazevic N. Antimicrobial activity of juniper berry essential oil (J uniperus communis L., Cupressaceae). Acta Pharm 2005;55:417-422.
45. Ali BH, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytother Res 2003;17:299-305.
46. Silva BA, Ferreres F, Malva J O, Dias ACP. Phytochemical and antioxidant characterization of Hypericum perf oratum alcoholic extracts. Food Chem 2005;90:157-167.
47. Lefebvre T, Foster BC, Drouin CE, Krantis A, Livesey JF, Jordan SA. In vitro activity of commercial valerian root extracts against human cytochrome P450 3A4. J Pharm Pharmaceut Sci 2004;72:265-273.
48. Akhondzadeh S, Noroozian M, Mohammadi M, Ohadinia S, Jamshidi A, Khani M. Melissa off icinalis extract in the treatment of patients with mild to moderate Alzheimers disease: a double blind, randomised, placebo controlled trial. J Neurol Neurosurg Psychiatry 2003;74:863-866.
49. Ren W, Qiao Z, Wang H, Zhu L, Zhang L. Flavonoids: promising anticancer agents. Med Res Rev 2003;23:519-534.
50. Salminen A, Lehtonen M, Suuronen T, Kaarniranta K, Huuskonen J. Terpenoids: natural inhibitors of NF-B signaling with anti-inflammatory and anticancer potential. Cell Mol Life Sci 2008;65:2979-2999.
51. Rabi T, Bishayee A. Terpenoids and breast cancer chemoprevention. Breast Cancer Res Treat 2009;115:223- 239.
52. Vandenbogaerde AL, de Witte PA. Hypericin as a natural photosensitizer with cytotoxic and antitumor effects. Phytother Res 1996;10:S150-S152.
53. Roscetti G, Franzese O, Comandini A, Bonmassar E. Cytotoxic activity of Hypericum perf oratum L. on K562 erythroleukemic cells: differential effects between methanolic extract and hypericin. Phytother Res 2004;18:66-72.

APA	GÖKBULUT A, YAŞAR M, Baran Y (2015). A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. , 118 - 126.
Chicago	GÖKBULUT Aysun Adan,YAŞAR Mustafa,Baran Yusuf A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. (2015): 118 - 126.
MLA	GÖKBULUT Aysun Adan,YAŞAR Mustafa,Baran Yusuf A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. , 2015, ss.118 - 126.
AMA	GÖKBULUT A,YAŞAR M,Baran Y A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. . 2015; 118 - 126.
Vancouver	GÖKBULUT A,YAŞAR M,Baran Y A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. . 2015; 118 - 126.
IEEE	GÖKBULUT A,YAŞAR M,Baran Y "A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells." , ss.118 - 126, 2015.
ISNAD	GÖKBULUT, Aysun Adan vd. "A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells". (2015), 118-126.

APA	GÖKBULUT A, YAŞAR M, Baran Y (2015). A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. Turkish Journal of Hematology, 32(2), 118 - 126.
Chicago	GÖKBULUT Aysun Adan,YAŞAR Mustafa,Baran Yusuf A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. Turkish Journal of Hematology 32, no.2 (2015): 118 - 126.
MLA	GÖKBULUT Aysun Adan,YAŞAR Mustafa,Baran Yusuf A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. Turkish Journal of Hematology, vol.32, no.2, 2015, ss.118 - 126.
AMA	GÖKBULUT A,YAŞAR M,Baran Y A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. Turkish Journal of Hematology. 2015; 32(2): 118 - 126.
Vancouver	GÖKBULUT A,YAŞAR M,Baran Y A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells. Turkish Journal of Hematology. 2015; 32(2): 118 - 126.
IEEE	GÖKBULUT A,YAŞAR M,Baran Y "A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells." Turkish Journal of Hematology, 32, ss.118 - 126, 2015.
ISNAD	GÖKBULUT, Aysun Adan vd. "A novel natural Product, KL-21, Inhibits Proliferation and Induces Apoptosis in Chronic Lymphocytic Leukemia Cells". Turkish Journal of Hematology 32/2 (2015), 118-126.