Adipojenik mezenkimal kök hücrelerden farklılaştırılan doğal öldürücü hücrelerin over kanseri üzerindeki in vitro etkisinin incelenmesi
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Dosyalar
Tarih
2019
Yazarlar
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Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Over kanseri, kadınlarda kanserden ölümlerde ikinci sırada yer alan en yaygın jinekolojik
kanserlerdendir. Geç dönemde belirti vermesi nedeni ile hastaların %70’i teşhis konulduktan sonra beş yıl içinde hayatını kaybeder. Mezenkimal kök hücreler (MKH), multipotent erişkin kök hücre tipidir ve stromal kökenli oldukları için “destek hücresi” özelliğI de taşırlar. Bu hücreler özellikle son dönemlerde doku mikroçevresi için önemli olmaları ve immün sistem üzerine baskılayıcı özellik taşımalarından dolayı dikkat çekmektedirler. Kemik iliği ve adipoz doku gibi birçok dokudan elde edilebilirler ve çoğaltılmaya elverişli olduklarından in vitro ortamda pasajlanarak çoğaltılabilen hücrelerdir. Genellikle cerrahi işlemlerle kolay ve bol miktarda elde edilebilen adipoz doku, zengin bir MKH kaynağıdır. Doğal öldürücü (NKNatural killer) hücreler malignansi ve enfeksiyon durumunda konak immünitesinin kritik mediyatörleridir. NK hücrelerinin güçlü antitümör yanıtlara aracılık ettiği preklinik fare ve klinik insan çalışmalarında gösterilmiştir. Fonksiyonel NK hücrelerinin aktivasyonu; kemokin ve sitokin üretimi (özellikle INFγ hedef hücrelerde güçlü düzenleyici etkiler gösterir) yanısıra NK hücresi aracılı sitotoksisiteye (perforin ve granzim B gibi litik moleküllerin salınımı ile) yol açarak malign hücreleri apoptoza sürükleyerek yokeder. Bu projede adipojenik mezenkimal kök hücreler (ADMKH)’den farklılaştırılan NK hücrelerinin HeyA8 over kanser hücreleri üzerindeki etkisi in vitro olarak incelenmiştir. Çalışma kapsamında, NK hücresi diferansiyasyonu yapılmıştır ve karakterize edilmiştir. Karakterizasyon için, NK hücrelerine özgü CD314 antikoru kullanılarak flow sitometri analizi yapılmış ve NK özelliğine sahip hücreler, HeyA8 hücreleri ile ko-kültüre edilmiştir. Kokültür ortamında NK hücrelerinin varlığı ve sitotoksik etkisi belirlenmiştir. NK hücrelerinin, HeyA8 hücreleri ile ko-kültür yapldığında bu hücrelerde apoptozu tetiklediği gösterilmiştir. NK hücreleri, over kanser hücreleri üzerinde yapılan bu in vitro çalışmanın sonuçlarına göre, immünoterapötik yaklaşım açısından verimli ve etkin bir hücresel tedavi modeli oluşturmaktadır.
Ovarian cancer is one of the most common cancers which takes/represents the second place among deaths from cancer in women. Since it shows its symptoms at a later stage, 70% of the patients pass away within the five years after diagnosis. Mesenchymal stem cells (MSC) are multipotent adult stem cells and because they originate from stroma, they also show the characteristics of “supporting cells”. These cells recently gained attention due to their importance for tissue microenvironment and their suppressive effects on immune system. They could be obtained from various tissues, such as bone marrow and adipose tissue and since they represent a favorable cell type for proliferation, they can be expanded in in vitro conditions via subculturing. Adipose tissue, which could generally be obtained easily and in massive amounts via surgical procedures, is a rich source of MSCs. Natural Killer cells are critical mediators of host immunity in cases of cellular malignancy and infection. It was shown with both preclicinal and clinical studies on mice and humans, respectively that, Natural Killer cells mediate strong antitumoral responses. Activation of functional NK cells; besides causing the release of chemokines and cytokines (especially INFγ shows strong regulatory effects on targeted cells), it leads to NK cell-mediated cytotoxicity (via releasing lytic molecules such as perforine and granzyme B) which in turn kills malignant cells through leading to apoptosis. In this project, in vitro effect of NK cells differentiated from adipogenic mesenchymal stem cells (AD-MSCs) on HeyA8 ovarian cancer cells was investigated. In scope of the study, differentiation to NK cells was done and characterized via analysing NK cell-specific antibody-CD314 by flow cytometry. The cells showing characteristics of NK cells were determined and then, co-cultured with HeyA8 cells. Presence of NK cells and their cytotoxic effect was determined in co-culturing media. It was shown that these cells stimulated apoptosis when they were co-cultured with HeyA8 cells. Based on the results obtained in this in vitro study on ovarian cancer cells, NK cells represent an efficient and effective cellular treatment model from the perspective of immunotherapeutic approach.
Ovarian cancer is one of the most common cancers which takes/represents the second place among deaths from cancer in women. Since it shows its symptoms at a later stage, 70% of the patients pass away within the five years after diagnosis. Mesenchymal stem cells (MSC) are multipotent adult stem cells and because they originate from stroma, they also show the characteristics of “supporting cells”. These cells recently gained attention due to their importance for tissue microenvironment and their suppressive effects on immune system. They could be obtained from various tissues, such as bone marrow and adipose tissue and since they represent a favorable cell type for proliferation, they can be expanded in in vitro conditions via subculturing. Adipose tissue, which could generally be obtained easily and in massive amounts via surgical procedures, is a rich source of MSCs. Natural Killer cells are critical mediators of host immunity in cases of cellular malignancy and infection. It was shown with both preclicinal and clinical studies on mice and humans, respectively that, Natural Killer cells mediate strong antitumoral responses. Activation of functional NK cells; besides causing the release of chemokines and cytokines (especially INFγ shows strong regulatory effects on targeted cells), it leads to NK cell-mediated cytotoxicity (via releasing lytic molecules such as perforine and granzyme B) which in turn kills malignant cells through leading to apoptosis. In this project, in vitro effect of NK cells differentiated from adipogenic mesenchymal stem cells (AD-MSCs) on HeyA8 ovarian cancer cells was investigated. In scope of the study, differentiation to NK cells was done and characterized via analysing NK cell-specific antibody-CD314 by flow cytometry. The cells showing characteristics of NK cells were determined and then, co-cultured with HeyA8 cells. Presence of NK cells and their cytotoxic effect was determined in co-culturing media. It was shown that these cells stimulated apoptosis when they were co-cultured with HeyA8 cells. Based on the results obtained in this in vitro study on ovarian cancer cells, NK cells represent an efficient and effective cellular treatment model from the perspective of immunotherapeutic approach.
Açıklama
Anahtar Kelimeler
Over Kanseri, İmmun Sistem, İmmunoterapi, NK Hücreleri, Ovarian Cancer, Immune System, Immunotherapy, NK Cells
