Anti kanser ajanlarının mikroorganizmalardan izolasyonu ve etkinliklerinin araştırılması
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Tarih
2019
Yazarlar
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Cilt Başlığı
Yayıncı
Ege Üniversitesi, Fen Bilimleri Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Dünyada her yıl 17 milyon kişi kansere yakalanmakta ve 8 milyonu kanser nedeniyle kaybedilmektedir. 2015 yılında, Dünya Sağlık Örgütü'nün verilerine göre, 172 ülkeden 91 ülkede erken yaşta ölümlerin birinci ve ikinci sebebi, 22 ülkede ise üçüncü ve dördüncü sebebi kanserdir. Kanserden ölümlerin başında akciğer kanseri gelmektedir. Tek başına akciğer kanseri, iskemik kalp hastalığı, inme ve diabetes mellitus'tan sonraki dördüncü global ölümcül hastalıktır. Tez kapsamında, kanser tedavisinde kullanılabilecek endofitlerden sekonder metaboliterinin saflaştırılması ve kanser tedavi potansiyellerinin in vitro ve in vivo koşullarda araştırılması hedeflenmiştir. Punica granatum' ın çiçekleri, meyveleri, yaprakları, dalları ve köklerinde yer alan mantarların izolasyonu yapıldı. İzole edilmiş endofitik mantarların, ürettikleri ekzoenzimler olan üreaz, jelatinaz, L-asparaginazve L-glutaminaz aktiviteleri tarandı. Üreaz (kök endofitleri: 5 (-); dal endofitleri: 3 (-); Meyve endofitleri: 1 (-), 1 (+)); jelatinaz (kök endofitleri: 6 (+)); L-asparaginaz: (kök endofitleri: 6 (+)), L-glutaminaz (kök endofiti: 5 (+), dal endofiti: 1 (+) olarak belirlendi. İzole edilen türün analizi yapılarak Aspergillus niger'in yeni bir türü tanımlandı. Aspergillus niger karmali'nin "pepton ile desteklenmiş pirinç" üreme ortamında fermentasyonu yapılarak kuru kolon vakumlu kromatografi ile fraksiyonlandı. Saflaştırma işlemlerinin ardından üretilen endofitin sekonder metabolit. 1H NMR, 13C NMR, 31P NMR ve LC-MS içeren spektroskopik analizleri gerçekleştirilerek C, H, O, P elementel bileşimi ve molekül kütlesi belirlenerek karakterize edildi. Endofit bileşiğin in vitro koşullarda kanser tedavi potansiyeli, over kanser hücresi SKOV3 ve akciğer kanser hücresi A549 kullanılarak sitotoksisite tayini ile belirlendi. Hücre kültürü denemeleri sonucunda 24.saatte IC50 değerleri SKOV3 hücresi için 1.08 µg/ml iken A549 hücresi için ise 11.7 µg/ml olarak saptanmıştır. Tez kapsamında saflaştırılan endofiti bileşiğin kanser tedavi potansiyelinin SKOV3 hücresi üzerinde daha etkili olduğu görülmektedir. in vivo koşullarda etkinliğinin belirlenmesi amacıyla biyouyumluluk testleri gerçekleştirilmiş ve biyouyumlu olduğu belirlenmiştir.
Every year 17 million new cases of cancer are diagnosed and 8 million people die due to cancer. In 2015, according to estimations of the World Health Organization (W.H.O.), cancer is the first or second cause of premature deaths in 91 countries out of 172. Additionally, cancer is the third or fourth cause of premature deaths in 22 countries. Lung cancer is the leading cause of cancer deaths. Lung cancer alone is the fourth global deadly disease after ischemic heart disease, stroke and diabetes mellitus. The aim of the thesis is to purify the secondary metabolites of endophytes which can be used in cancer treatment and to investigate the cancer treatment potentials in vitro and in vivo. Endophytic fungi have been isolated from flowers, fruits, leaves, twigs and roots of Punica granatum. The so isolated endophytes were screened for the production of exoenzymes including urease, gelatinase, L-asparaginase, and L-glutaminase and the following results were recorded: Urease (root endophytes: 5 (-); twig endophytes: 3 (-); Fruit endophytes: 1 (-), 1 (+)); gelatinase (root endophytes: 6 (+)); L-asparaginase: (root endophytes: 6 (+)), L-glutaminase (root endophyte: 5 (+), twig endophyte: 1 (+)). A new strain of Aspergillus niger i.e. Aspergillus niger karmalı. was identified by analysis of the isolated species. Aspergillus niger karmali, was grown on "rice supplemented with peptone" in a solid-state fermentation. The culture extract was fractionated using Dry Column Vacuum Chromatography (DCVC). Spectroscopic analyses of the sample including 1H NMR, 13C NMR, 31P NMR, LC-MS of the sample allowed for the determination of its elemental composition to be C,H,O,P with a molecular weight of 462. The cancer treatment potential of the endophytic compound in vitro, was determined as its cytotoxicity against ovarian cancer cell SKOV3 and lung cancer cell A549. As a result of cell culture experiments, IC50 values were determined as 1.08 µg / ml for SKOV3 cell and 11.7 µg / ml for A549 cell at 24 hours. The cancer treatment potential of the purified endophyte compound was found to be more effective on SKOV3 cell. Biocompatibility tests were performed in order to determine its effectiveness in vivo conditions and it was determined to be biocompatible.
Every year 17 million new cases of cancer are diagnosed and 8 million people die due to cancer. In 2015, according to estimations of the World Health Organization (W.H.O.), cancer is the first or second cause of premature deaths in 91 countries out of 172. Additionally, cancer is the third or fourth cause of premature deaths in 22 countries. Lung cancer is the leading cause of cancer deaths. Lung cancer alone is the fourth global deadly disease after ischemic heart disease, stroke and diabetes mellitus. The aim of the thesis is to purify the secondary metabolites of endophytes which can be used in cancer treatment and to investigate the cancer treatment potentials in vitro and in vivo. Endophytic fungi have been isolated from flowers, fruits, leaves, twigs and roots of Punica granatum. The so isolated endophytes were screened for the production of exoenzymes including urease, gelatinase, L-asparaginase, and L-glutaminase and the following results were recorded: Urease (root endophytes: 5 (-); twig endophytes: 3 (-); Fruit endophytes: 1 (-), 1 (+)); gelatinase (root endophytes: 6 (+)); L-asparaginase: (root endophytes: 6 (+)), L-glutaminase (root endophyte: 5 (+), twig endophyte: 1 (+)). A new strain of Aspergillus niger i.e. Aspergillus niger karmalı. was identified by analysis of the isolated species. Aspergillus niger karmali, was grown on "rice supplemented with peptone" in a solid-state fermentation. The culture extract was fractionated using Dry Column Vacuum Chromatography (DCVC). Spectroscopic analyses of the sample including 1H NMR, 13C NMR, 31P NMR, LC-MS of the sample allowed for the determination of its elemental composition to be C,H,O,P with a molecular weight of 462. The cancer treatment potential of the endophytic compound in vitro, was determined as its cytotoxicity against ovarian cancer cell SKOV3 and lung cancer cell A549. As a result of cell culture experiments, IC50 values were determined as 1.08 µg / ml for SKOV3 cell and 11.7 µg / ml for A549 cell at 24 hours. The cancer treatment potential of the purified endophyte compound was found to be more effective on SKOV3 cell. Biocompatibility tests were performed in order to determine its effectiveness in vivo conditions and it was determined to be biocompatible.
Açıklama
Anahtar Kelimeler
Endofitler, Mantarlar, Katı Hal Fermentasyonu, Jelatinaz, L-asparaginaz, L-glutaminaz, Üreaz, Amilaz, Punica Granatum, A549, Scov-3, Antikanser, Endophytes, Fungi, Solid State Fermentation, Gelatinase, L-asparaginase, L-glutaminase, Urease, Amylase, Punica Granatum, A549, SKOV-3, Anticancer
