Association Between Anti-Proliferative Activity of Evernia Prunastri with the Cellular Apoptotic Pathway
Abstract
Evernia prunastri (L.) Ach, belonging to Parmeliaceae, is an
important lichen species in Turkey. Previous studies was reported that E. prunastri have significant
antioxidant, antimicrobial and anticancer compounds in its structure. Although
antiproliferative effects of E. prunastri
are determined in some types of cancer, there is little information about the
pathway of this activity. Accordingly, we aimed to determine the association
between anti-proliferative activities of E.
prunastri extracts with the cellular apoptotic pathway. Determination of
cell viability was assessed by MTT assay. The xCELLigence system was used to
monitor cell proliferation. To reveal induction apoptosis, expression of
apoptotic pathway proteins, F-actin staining, ANNEXIN
V/propodium iodide (PI) staining was used. As a result, E. prunastri MeOH extracts were found to have anti-proliferative activities
especially in high doses (p<0.05 for 1, 5 and 25 µg ml-1;
p<0.01 for 125 µg ml-1). Also, the extracts were found to be not inducing apoptosis in flow cytometric
measurements. In addition, there
was no significant decrease or increase in protein expressions of apoptotic
pathway proteins. Lastly, density of actin filaments was not affected. MeOH extract of E. prunastri was found to have
anti-proliferative activities, yet these anti-proliferative activities were not
related to cellular apoptotic pathways.
References
- [1]. R.L. Siegel, K.D. Miller, A. Jemal. Cancer statistics. Cancer J Clin. 1986, 65: 5-29.
- [2]. American Cancer Society, 2016. Breast Cancer Facts & Figures 2015-2016. Atlanta: American Cancer Society, Inc. Avaliable at: https://www.cancer.org/content/dam/cancer-org/research/cancer-facts-and statistics/breast-cancer-facts-and-figures/breast-cancer-facts-and-figures-2015-2016.pdf.
- [3]. M.J. Bak, S.D. Gupta, J. Wahler, N. Suh. Role of dietary bioactive natural products in estrogen receptor-positive breast cancer. Semin Cancer Biol. 2016,41,170-191.
- [4]. G. Shrestha, L.L. St Clair. Lichens: a promising source of antibiotic and anticancer drugs. Phytochem Rev. 2013, 12, 229-244.
- [5]. K.O. Vartia, Antibiotics in lichens. In: Ahmadjian V, Hale ME (ed.), The Lichens, New York, Academic Press, 1973, pp. 547-561
- [6]. V. Shukla, G.P. Joshi, M.S.M. Rawat. Lichens as a potential natural source of bioactive compounds: a review. Phytochem Rev. 2010, 9, 303-314.
- [7]. V. Cardile, A.C.E.Graziano, R. Avola, M. Piovano, A. Russo. Potential anticancer activity of lichen secondary metabolite physodic acid. Chem Biol Interact. 2017, 263, 36-45.
- [8]. M. Bessadottir, E.A. Skuladottir, S. Gowan, S. Eccles, S. Omarsdottir, H.M. Ogmundsdottir. Effects of anti-proliferative lichen metabolite, protolichesterinic acid on fatty acid synthase, cell signalling and drug response in breast cancer cells Phytomedicine. 2014, 21, 1717-1724.
- [9]. Russo, S. Caggia, M. Piovano, J. Garbarino, V. Cardile. Effect of vicanicin and protolichesterinic acid on human prostate cancer cells: role of Hsp70 pro- tein. Chem Biol Interact. 2012, 195, 1-10 .
- [10]. M. Backorova, R. Jendzelovsky, M. Kello, M. Backor, J. Mikes, P. Fedorocko. Lichen secondary metabolites are responsible for induction of apoptosis in HT-29 and A2780 human cancer cell lines Toxicol in Vitro. 2012, 26, 462-468.
- [11]. M. Kosanic, N. Manojlovic, S. Jankovic, T. Stanojkovic, B. Rankovic. Evernia prunastri and Pseudoevernia furfuraceae lichens and their major metabolites as antioxidant, antimicrobial and anticancer agents. Food Chem Toxicol. 2013, 53, 112-118.
- [12]. T. Mitrovic, S. Stamenkovic, V. Cvetkovic, S.Tosic, M. Stankovic, I. Radojevic, O. Stefanovic, L. Comic, D. Dacic, M. Curcic, S. Markovic. Antioxidant, Antimicrobial and Antiproliferative Activities of Five Lichen Species. Int. J Mol Sci. 2011, 12, 5428-5448.
- [13]. D. Triggiani, D. Ceccarelli, A. Tiezzi, T. Pisani, S. Munzi, C. Gaggi, S. Loppi. Antiproliferative activity of lichen extracts on murine myeloma cells. Biologia. 2009, 64, 59-62.
- [14]. S. Elmore. Apoptosis: A review of programmed cell death. Toxicol. Pathol. 2007, 35, 495-516.
- [15]. Vermes, C. Haanen, H. Steffens-Nakken, C. Reutellingsperger. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods. 1995, 184, 39-51.
- [16]. B. Burlando, E. Ranzato, A. Volante, G. Appendino, F. Pollastro, L. Verotta. Antiproliferative effects on tumour cells and promotion of keratinocyte wound healing by different lichen compounds. Planta medica. 2009, 75, 607-613.
- [17]. Russo, M. Piovano, L. Lombardo, J. Garbarino, V. Cardile. Lichen metabolites prevent UV light and nitric oxide-mediated plasmid DNA damage and induce apoptosis in human melanoma cells. Life Sci. 2008, 83, 468-474.
- [18]. M. Backorova, M. Backor, J. Mikes, R. Jendzelovski, P. Fedorocko. Variable responses of different human cancer cells to the lichen compounds parietin, atranorin, usnic acid and gyrophoric acid Toxicol in Vitro, 2011, 25, 37-44.
- [19]. S. Kothakota, T. Azuma, C. Reinhard, A. Klippel, J. Tang, K. Chu, T.J. McGarry, M.W. Kirschner, K. Koths, D.J. Kwiatkowski, L.T. Williams (1997). Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis. Science. 1997, 278, 294-298.
Abstract
Parmeliaceae’ye ait olan
Evernia prunastri (L.) Ach, Türkiye
yetişen önemli bir liken türüdür. önceki çalışmalarda E. Prunastri’nin yapısında bulanan kimyasallardan dolayı
antioksidan, antimikrobiyal ve antikanser aktivitelere sahip oldukları
belirtilmiştir. E. Prunastri’nin bazı
kanser hücre tiplerinde antiproliferatif
aktiviteleri bilinmekle birlikte bu aktivitenin yolağı hakkında bilgilerimiz
oldukça sınırlıdır. Bundan dolayı, çalışmamızda E. Prunastri ekstraktının antiproliferatif etkileri ve hücresel
apoptotik yolak arasında ilişkinin belirlenmesi amaçlanmıştır. Bu amaç için,
meme kanseri hücrelerini E. Prunastri ekstraktlarına
maruz bırakılmış ve hücre canlılığı MTT boyama ve xCELLigence system ile
belirlenmiştir. Annexine V/PI boyama ile Apoptozun indüksiyonu, westernblot ile
apoptotik proteinlerin ekspresyonu ve floresan boyama ile aktin flamentlerinin
hücresel yoğunluğu belirlenmiştir. E.
Prunastri MeOH ekstraktının yüksek dozlarda antiproliferatif etkiler
göstermiştir (1, 5 ve 25 µg ml-1
için p<0.05 ; 125 µg ml-1 için p<0.01). Ancak ekstrakt
uygulaması sonrasında apoptozun indüklenmediği ve apoptotik protein
seviyelerinin değişmediği belirlenmiştir. Ayrıca F-aktin seviyesininde
değişmediği gözlemlenmiştir. Sonuç olarak, E. Prunastri MeOH ekstraktının meme kanseri hücreleri üzerine
antiproliferatif bir etkiye sahip olduğu ancak bu etkinin apoptotik yolakla
ilişkili olmadığı düşünülmektedir.
References
- [1]. R.L. Siegel, K.D. Miller, A. Jemal. Cancer statistics. Cancer J Clin. 1986, 65: 5-29.
- [2]. American Cancer Society, 2016. Breast Cancer Facts & Figures 2015-2016. Atlanta: American Cancer Society, Inc. Avaliable at: https://www.cancer.org/content/dam/cancer-org/research/cancer-facts-and statistics/breast-cancer-facts-and-figures/breast-cancer-facts-and-figures-2015-2016.pdf.
- [3]. M.J. Bak, S.D. Gupta, J. Wahler, N. Suh. Role of dietary bioactive natural products in estrogen receptor-positive breast cancer. Semin Cancer Biol. 2016,41,170-191.
- [4]. G. Shrestha, L.L. St Clair. Lichens: a promising source of antibiotic and anticancer drugs. Phytochem Rev. 2013, 12, 229-244.
- [5]. K.O. Vartia, Antibiotics in lichens. In: Ahmadjian V, Hale ME (ed.), The Lichens, New York, Academic Press, 1973, pp. 547-561
- [6]. V. Shukla, G.P. Joshi, M.S.M. Rawat. Lichens as a potential natural source of bioactive compounds: a review. Phytochem Rev. 2010, 9, 303-314.
- [7]. V. Cardile, A.C.E.Graziano, R. Avola, M. Piovano, A. Russo. Potential anticancer activity of lichen secondary metabolite physodic acid. Chem Biol Interact. 2017, 263, 36-45.
- [8]. M. Bessadottir, E.A. Skuladottir, S. Gowan, S. Eccles, S. Omarsdottir, H.M. Ogmundsdottir. Effects of anti-proliferative lichen metabolite, protolichesterinic acid on fatty acid synthase, cell signalling and drug response in breast cancer cells Phytomedicine. 2014, 21, 1717-1724.
- [9]. Russo, S. Caggia, M. Piovano, J. Garbarino, V. Cardile. Effect of vicanicin and protolichesterinic acid on human prostate cancer cells: role of Hsp70 pro- tein. Chem Biol Interact. 2012, 195, 1-10 .
- [10]. M. Backorova, R. Jendzelovsky, M. Kello, M. Backor, J. Mikes, P. Fedorocko. Lichen secondary metabolites are responsible for induction of apoptosis in HT-29 and A2780 human cancer cell lines Toxicol in Vitro. 2012, 26, 462-468.
- [11]. M. Kosanic, N. Manojlovic, S. Jankovic, T. Stanojkovic, B. Rankovic. Evernia prunastri and Pseudoevernia furfuraceae lichens and their major metabolites as antioxidant, antimicrobial and anticancer agents. Food Chem Toxicol. 2013, 53, 112-118.
- [12]. T. Mitrovic, S. Stamenkovic, V. Cvetkovic, S.Tosic, M. Stankovic, I. Radojevic, O. Stefanovic, L. Comic, D. Dacic, M. Curcic, S. Markovic. Antioxidant, Antimicrobial and Antiproliferative Activities of Five Lichen Species. Int. J Mol Sci. 2011, 12, 5428-5448.
- [13]. D. Triggiani, D. Ceccarelli, A. Tiezzi, T. Pisani, S. Munzi, C. Gaggi, S. Loppi. Antiproliferative activity of lichen extracts on murine myeloma cells. Biologia. 2009, 64, 59-62.
- [14]. S. Elmore. Apoptosis: A review of programmed cell death. Toxicol. Pathol. 2007, 35, 495-516.
- [15]. Vermes, C. Haanen, H. Steffens-Nakken, C. Reutellingsperger. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods. 1995, 184, 39-51.
- [16]. B. Burlando, E. Ranzato, A. Volante, G. Appendino, F. Pollastro, L. Verotta. Antiproliferative effects on tumour cells and promotion of keratinocyte wound healing by different lichen compounds. Planta medica. 2009, 75, 607-613.
- [17]. Russo, M. Piovano, L. Lombardo, J. Garbarino, V. Cardile. Lichen metabolites prevent UV light and nitric oxide-mediated plasmid DNA damage and induce apoptosis in human melanoma cells. Life Sci. 2008, 83, 468-474.
- [18]. M. Backorova, M. Backor, J. Mikes, R. Jendzelovski, P. Fedorocko. Variable responses of different human cancer cells to the lichen compounds parietin, atranorin, usnic acid and gyrophoric acid Toxicol in Vitro, 2011, 25, 37-44.
- [19]. S. Kothakota, T. Azuma, C. Reinhard, A. Klippel, J. Tang, K. Chu, T.J. McGarry, M.W. Kirschner, K. Koths, D.J. Kwiatkowski, L.T. Williams (1997). Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis. Science. 1997, 278, 294-298.
Details
| Journal Section | Articles |
|---|---|
| Authors | |
| Publication Date | September 30, 2017 |
| Submission Date | March 9, 2017 |
| Acceptance Date | June 29, 2017 |
| Published in Issue | Year 2017 |