İzzet İSLAM, Hande AYTUĞ, İbrahim BULDUK, Funda KARABAĞ ÇOBAN, Alper KARAGÖZ

insan akciğer hücre hatlarında noskapinin antianjiogenik ve apopitotik etkileri

Giriş ve Amaç: Günümüzün en önemli sağlık sorunlarından biri olan akciğer kanseri hem kadınlarda hem de erkeklerde en sık ölüme neden olan kanser türüdür. İnsan alveolar karsinom hücrelerinden kaynaklanan A549 akciğer hücreleri, tip II alveolar hücre fenotipiyle eşleşmesine rağmen, insan primer alveolar epitel hücrelerinin sahip olduğu birçok özelliğe de sahiptir. Noskapin, haşhaş somniferum (afyon) içindeki bileşenlerden oluşur. İlk olarak 1817'de Papaver somniferum'dan (afyon) izole edilmiştir. Morfinden sonra afyon bitkisinde (toplam bileşimin %10'una kadar) bulunan en bol opioidlerden oluşur. Narcotine, Nectodon, Nospen, Anarcotine ve (arkaik) Opiane olarak da bilinir ve S, R stereokimyasına sahip bir (-) izomerdir (fitalid-karbonda S stereokimyası ve izokinolin-karbonda R). Noskapin yapısal ve kimyasal olarak morfin, kodein, tebain, papaverin ve narsein gibi diğer afyon alkaloidlerinden farklıdır. Bu çalışmanın amacı, insan akciğer kanseri (A549) hücrelerinde noskapin'in apoptotik ve antianjiogenik etkilerinin araştırılmasıdır. Gereç ve Yöntemler: Çalışmada 10 ppm, 15 ppm, 20 ppm, 25 ppm, 40 ppm, 50 ppm, 60 ppm, 65 ppm, 70 ppm, 75 ppm, 80 ppm, 90 ppm ve 100 ppm konsantrasyonları kullanılmıştır. Bulgular: Canlılık değerlerinin çoğalma deneyi oranı. Düştüğü gözlendi ve LD50 değeri 40 ppm olarak belirlendi. Kullanılan istatistiksel analiz: Analizlerin istatistiksel analizi SPSS Statistics 20,0 programı kullanılarak yapıldı. Bulgular: VEGF değerleri 20 ppm, 40 ppm ve 80 ppm noskapin konsantrasyonlarında kontrol grubuna göre azaldı. 80 ppm'de PARP değerlerinde önemli bir düşüş oldu. Sonuç: Bulgular sonucunda; Noskapin konsantrasyonlarının 10 ppm, 20 ppm olduğu düşünülmektedir. 40 ppm, VEGF seviyelerini düşürerek anjiyogenezi azaltabilir ve metastazı önleyebilir. PARP seviyeleri tüm noskapin konsantrasyonlarında azaldı, ancak en önemli fark 80 ppm'de görüldü.

Anahtar Kelimeler:

A549, Noscapine, PARP, Papaver Somniferum, VEGF

Apoptotic And Antiangiogenic Effects of Noscapine In Human Lung Cancer (A549) Cells

Objective: Lung cancer, one of the most important health problems today, is the most common type of cancer that causes death in both women and men. Although A549 lung cells originating from human alveolar carcinoma cells match the type II alveolar cell phenotype, it also has many characteristics that human primary alveolar epithelial cells have. Noscapine consists of the components in the poppy somniferum (opium). It was first isolated from Papaver somniferum (opium) in 1817. It consists of the most abundant opioids found in the opium plant (up to 10% of the total composition) after morphine. It is also known as Narcotine, Nectodon, Nospen, Anarcotine and (archaic) Opiane, and S is an (-) isomer with R stereochemistry (S stereochemistry in phthalid-carbon and R in isoquinoline-carbon). Noscapine is structurally and chemically different from other opium alkaloids such as morphine, codeine, thebaine, papaverine and narcein.The aim of the study the ınvestıgatıon of the apoptotıc and antıangıogenıc effects of noscapine ın human lung cancer (A549) cells. Materials and Methods: In the study, concentrations of 10 ppm, 15 ppm, 20 ppm, 25 ppm, 40 ppm, 50 ppm, 60 ppm, 65 ppm, 70 ppm, 75 ppm, 80 ppm, 90 ppm and 100 ppm were used in the proliferation experiment proportion of vitality values. It was observed that it decreased and the LD50 value was determined as 40 ppm. Statistical analysis used: Statistical analysis of the analyzes was performed using SPSS Statistics 20,0 program. Results: VEGF values decreased at 20 ppm, 40 ppm and 80 ppm noscapine concentrations compared to the control group. There was a significant decrease in PARP values at 80 ppm. Conclusions: As a result of the findings; It is thought that concentrations of noscapine 10 ppm, 20 ppm and 40 ppm can reduce angiogenesis and prevent metastasis by lowering VEGF levels. PARP levels decreased at all noscapine concentrations but the most significant difference was seen at 80 ppm noscapine concentration.

Keywords:

A549, Noscapine, PARP, Papaver Somniferum, VEGF,

PDF

___

Gelband, H, Sankaranarayanan, R, Gauvreau, C.L, Horton, S, Anderson, B.O, Bray, F, et al., Costs, affordability, and feasibility of an essential package of cancer control interventions in low-income and middle-income countries: key messages from Disease Control Priorities, The Lancet, 2016, 387(10033):2133-44.
Cairns, R.A, Mak, T.W, The current state of cancer metabolism, Nature Reviews Cancer, 2016, 16(10), 613. Levitsky, D.O, Dembitsky, V.M, Anti-breast cancer agents derived from plants, Natural products and bioprospecting, 2015, 5(1), 1-6.
Tagne, RS, Telefo, B.P, Nyemb, J.N, Yemele, D.M, Njina, S.N, Goka, S.M, et al. Anticancer and antioxidant activities of methanol extracts and fractions of some Cameroonian medicinal plants, Asian Pacific journal of tropical medicine, 2014, 7, S442-7.
Torre, L.A, Siegel, R.L, Ward, E.M, Jemal, A, Global cancer incidence and mortality rates and trends—an update, Cancer Epidemiology and Prevention Biomarkers, 2016, 25(1), 16-27.
Giard, D.J, Aaronson, S.A, Todaro, G.J, Arnstein, P, Kersey, J.H, Dosik, H, et al, In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors, Journal of the National Cancer Institute, 1973, 51(5):1417-23.
Lieber, M, Todaro, G, Smith, B, Szakal, A, Nelson‐Rees, W.A continuous tumor‐cell line from a human lung carcinoma with properties of type II alveolar epithelial cells, International journal of cancer, 1976, 17(1), 62-70.
Fang, R, Aust, A.E, Induction of ferritin synthesis in human lung epithelial cells treated with crocidolite asbestos, Archives of biochemistry and biophysics, 1997, 340(2), 369-75.
Shimizu, K, Endo, O, Goto, S, Sakoda, A, Ono, Y, Sakai, Y, Bioassay-based evaluation of toxicity of suspended particulate matter in humans: integrated uses of alveolar cells (A549) in air–liquid interface culture and hepatocarcinoma cells (Hep G2), Biochemical engineering journal, 2004, 22(1), 1-9.
Tian, D, Zhu, M, Li, J, Ma, Y, Wu, R, Cigarette smoke extract induces activation of β-catenin/TCF signaling through inhibiting GSK3β in human alveolar epithelial cell line, Toxicology letters, 2009, 187(1), 58-62.
Jiang, R.D, Shen, H, Piao, Y.J, The morphometrical analysis on the ultrastructure of A549 cells, Rom. Journal of Morphology Embryoly, 2010, 51, 663-7.
Bunn, P.A, Epidemiologic aspects of lung cancer, Cancer Journal of Clinics, 2000, 50, 7-33.
Önmez, H, Papaver somniferum bitkisinden elde edilen alkaloidlerin ekstraksiyonunda kullanılan çözücü ve metodların karşılaştırılması (Doctoral dissertation, Selçuk Üniversitesi Fen Bilimleri Enstitüsü), 2007.
Küçük, Y, Türkiye’nin Çeşitli Yörelerinde Yetiştirilen Haşhaş Bitkilerinde Alkaloidlerin Esktraksiyonu ve Ekstraksiyonların Susuz Fen Ortamlarda Özelliklerinin İncelenmesi, Basılmamış Doktora Tezi, Ankara Üniversitesi Bilimleri Enstitüsü, Ankara,1996.
Tanker, M, Tanker, N, Farmakognozi. Ankara: Ankara Üniversitesi Eczacılık Fakültesi Yayınları, Cilt 1-2, (66), 2003.
Ye, K, Ke, Y, Keshava, N, Shanks, J, Kapp, J.A, Tekmal, R.R, et al., Opium alkaloid noscapine is an antitumor agent that arrests metaphase and induces apoptosis in dividing cells, Proceedings of the National Academy of Sciences, 1998, 95(4):1601-6.
Ke, Y, Ye, K, Grossniklaus, HE, Archer, DR, Joshi, HC, Kapp, JA. Noscapine inhibits tumor growth with little toxicity to normal tissues or inhibition of immune responses. Cancer Immunology, Immunotherapy, 2000, 49(4-5):217-25.
Joshi, HC, Zhou, J. Noscapine and analogues as potential chemotherapeutic agents. Drug news & perspectives, 2000, 13(9), 543-6.
Gottesman, M.M, Pastan, I. Biochemistry of multidrug resistance mediated by the multidrug transporter, Annual review of biochemistry, 1993, 62(1), 385-427.
Kavallaris, M, Burkhart, C.A, Horwitz, S.B, Antisense oligonucleotides to class III β-tubulin sensitize drug-resistant cells to Taxol, British journal of cancer, 1999, 80(7), 1020-5.
Giannakakou, P, Sackett, D.L, Kang, Y.K, Zhan, Z, Buters, J.T, Fojo, T, et al., Paclitaxel-resistant human ovarian cancer cells have mutant β-tubulins that exhibit impaired paclitaxel-driven polymerization, Journal of Biological Chemistry, 1997, 272(27), 17118-25.
Verma, A.K, Bansal, S, Singh, J, Tiwari, R.K, Sankar, V.K, Tandon, V, et al., Synthesis and in vitro cytotoxicity of haloderivatives of noscapine, Bioorganic & medicinal chemistry, 2006, 14(19), 6733-6.
Zhou, J, Gupta, K, Yao, J, Ye, K, Panda, D, Giannakakou, P, et al., Paclitaxel-resistant human ovarian cancer cells undergo c-Jun NH2-terminal kinase-mediated apoptosis in response to noscapine, Journal of Biological Chemistry, 2002, 277(42), 39777-85.
Aneja, R, Zhou, J, Zhou, B, Chandra, R, Joshi, HC, Treatment of hormone-refractory breast cancer: apoptosis and regression of human tumors implanted in mice, Molecular cancer therapeutics, 2006, 5(9), 2366-77.
Landen, J.W, Lang, R, McMahon, S.J, Rusan, N.M, Yvon, A.M, Adams, A.W, et al., Noscapine alters microtubule dynamics in living cells and inhibits the progression of melanoma, Cancer research, 2002, 62(14), 4109-14.
Aneja, R, Ghaleb, A..M, Zhou, J, Yang, VW, Joshi, H.C, p53 and p21 determine the sensitivity of noscapine-induced apoptosis in colon cancer cells, Cancer research, 2007, 67(8), 3862-70.
Jackson, T, Chougule, M.B, Ichite, N, Patlolla, R.R, Singh, M, Antitumor activity of noscapine in human non-small cell lung cancer xenograft model, Cancer chemotherapy and pharmacology, 2008, 63(1), 117-26.
Rouleau, M, Patel, A, Hendzel, M.J, Kaufmann, S.H, Poirier, G.G, PARP inhibition: PARP1 and beyond, Nature reviews cancer, 2010, 10(4):293-301.
Zhu, G, Chang, P, Lippard, S.J. Recognition of platinum− DNA damage by poly (ADP-ribose) polymerase-1, Biochemistry, 2010, 49(29), 6177-83.
Quisbert-Valenzuela, E.O, Calaf, G.M, Apoptotic effect of noscapine in breast cancer cell lines, International journal of oncology, 2016, 48(6), 2666-74.
Chougule, M, Patel, A.R, Sachdeva, P, Jackson, T, Singh, M. Anticancer activity of Noscapine, an opioid alkaloid in combination with Cisplatin in human non-small cell lung cancer, Lung cancer, 2011, 71(3), 271-82.
Tian, X, Liu, M, Zhu, Q, Tan, J, Liu, W, Wang, Y, et al., Down-regulation of liver-intestine cadherin enhances noscapine-induced apoptosis in human colon cancer cells, Expert review of anticancer therapy, 2017, 17(9), 857-63.
Rabzia, A, Khazaei, M, Rashidi, Z, Khazaei, M.R, Synergistic anticancer effect of paclitaxel and noscapine on human prostate cancer cell lines. Iranian journal of pharmaceutical research, 2017, 16(4), 1432.
Elisa Cleaved PARP Kit, (2011), Katalog No: KHO0741, Invitrogen Corporation, Kalifornia, A.B.D. (www.invitrogen.com),
Kocak, C, Kocak, F.E, Ozturk, B, Tekin, G, Vatansev, H. Cytotoxic, anti-proliferative and apoptotic effects of noscapine on human estrogen receptor positive (MCF-7) and negative (MDA-MB-231) breast cancer cell lines, Bratislavske lekarske listy, 2020, 121(1), 43-50.
Kiechle, F.L, Zhang, X, Apoptosis: biochemical aspects and clinical implications, Clinica Chimica Acta, 2002, 326(1-2), 27-45.
Bursch, W, Karwan, A, Mayer, M, Dornetshuber, J, Fröhwein, U, Schulte-Hermann, R, et al., Cell death and autophagy: cytokines, drugs, and nutritional factors, Toxicology, 2008, 254(3), 147-57.
Shen, J.K, Du, H.P, Yang, M, Wang, Y.G, Jin, J, Casticin induces leukemic cell death through apoptosis and mitotic catastrophe, Annals of hematology, 2009, 88(8), 743-52.
Chougule, M, Patel, A.R, Sachdeva, P, Jackson, T, Singh, M. Anticancer activity of Noscapine, an opioid alkaloid in combination with Cisplatin in human non-small cell lung cancer, Lung cancer, 2011, 71(3), 271-82.
Konukoglu, D, Turhan, M.S. Anjiogenezin temel moleküler mekanizmaları ve tümör anjiogenezi, Cerrahpasa Tıp Dergisi, 2005, 36(1), 42-48.
Distler, J.H, Hirth, A, Kurowska-Stolarska, M, Gay, R.E, Angiogenic and angiostatic factors in the molecular control of angiogenesis, The Quarterly Journal of Nuclear Medicine and Molecular Imaging, 2003, 47(3),149.
Ferrara, N, Davis-Smyth, T. The biology of vascular endothelial growth factor. Endocrine reviews, 1997, 18(1), 4-25.
Ferrara, N, Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney international, 1999, 56(3),794-814.
Fox, S.B, Gatter, K.C, Haris, A.L, Tumor angiogenesis, Journal of Pathol, 1996, 179, 232-237.
Bloemendal, H.J, Logtenberg, T, Voest, E.E, New strategies in anti-vascular cancer therapy, European journal of clinical investigation, 1999, 29(9), 802-9.
Red-Horse, K, Ferrara, N, Imaging tumor angiogenesis, Journal of Clinical Investigation, 2006, 116(10), 2585-7.
O'Connell, J.B, Maggard, MA, Liu, JH, Etzioni, D.A, Rates of colon and rectal cancers are increasing in young adults, The American surgeon, 2003, 69(10), 866.
Johnson, I.T, Lund, E.K, Nutrition, obesity and colorectal cancer, Alimentary pharmacology & therapeutics, 2007, 26(2), 161-81.
Willett, W.C, Diet and cancer: an evolving Picture, Jama, 2005, 293(2), 233-4.