Transgenik salmonella immunotoksininin insan meme tümörü üzerinde antikanser etkinliği mekanizması
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Tarih
2024
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Kırıkkale Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Anti-kanser tedaviler özellikle de mevcut meme kanseri tedavileri iştahsızlık; enfeksiyonlara karşı direnç kaybı, halsizlik ve yorgunluk, kilo alımı, erken menopoz ve tümöre dirençli gelişme gibi kritik yan etkileri içermektedir. Bu sebeple; kanserin oluşturduğu potansiyel tehdidi önlemek için teşhis ve tedavide yeni yaklaşımlara ihtiyaç duyulmaktadır. Bu çalışmada; Pseudomonas Ekzotoksin (PE38) ile zayıflatılmış Salmonella Typhimurium'un insan meme adenokarsinomu (MCF-7) üzerindeki anti-tümör etkinliğinin makroskobik ve mikroskobik olarak değerlendirilmesi, uygulanan immunoterapotik tedavinin anti-tümöral etkinliğinin mekanizmasının araştırılması için için apoptoz (Kaspaz 3, 8 ve 9) ve piroptoz (Kaspaz 1, 11 ve GasderminE) varlığının gösterimi ve yolaklarının incelenmesi amaçlanmıştır. Sunulan bu çalışmada, MCF-7 hücre hattının in-vitro pasajlanmasının ardından Atimik Nude farelere implantasyonu gerçekleştirildi. Tümör oluşum ve çapı düzenli kontrol edilerek her birinin çapı 200mm3'ün üzerine çıktığında Pseudomonas ekzotoksin (PE38) ile TGF? ve PE38 ihtiva eden avirülent Salmonella Typhimurium (?ppGpp; relA, spoT and Glm mutant) ile immunoterapi uygulandı. Tedavi sonrası ötenazileri gerçekleştirilen farelerin sistemik nekropsileri yapılarak, oluşan tümöral dokular histopatolojik ve immunohistokimyasal incelemeler için fikse edildi. Buna göre histopatolojik olarak; kontrol grubu farelerde sadece adenokarsinom ilişkili tümöral hücrelere rastlanırken, tedavi uygulanan farelerin tümör dokularında neoplastik epitel hücrelere eşlik eden bakteri kümeleri, nekrotik ve apoptotik figürlere sahip ölü hücre popülasyonları ve yangı hücresi infiltrasyonuna rastlandı. Yine tedavi grubunda yoğun anti-kaspaz 3 ve 11 aktivitesine rastlanırken, orta şiddette anti- kaspaz 1,8 ve 9 aktivitelerine, hafif anti-Gasdermin E immunoreaktivitesine rastlandı. Buna karşın kontrol grubunda hiçbir immun-reaksiyona rastlanmadı. Sunulan bu çalışma ile, avirülent Salmonella Typhimurium' un anti-tümöral etkinliği kanıtlanmış ve bu anti-tümöral mekanizmada; nekroz, apoptoz ve piroptoz varlığı tümör hücrelerinin ölüm mekanizmaları olarak gösterilmiştir. Buna göre; meme kanseri tedavisinde başarılı bir alternatif yöntemin geliştirilmesi ve diğer kanser tedavileri için de temel oluşturacak etki mekanizmalarının belirlenmesi için temel niteliktedir. Anahtar Sözcükler: Anti-tümöral etkinlik, Apoptoz, İmmunohistokimya, MCF-7, Piroptoz, Salmonella Typhimurium.
Anti-cancer treatments, especially current breast cancer therapies, encompass critical side effects such as appetite loss, resistance to infections, weakness and fatigue, weight gain, early menopause, and development of resistance to tumors. Therefore, there is a need for new approaches in diagnosis and treatment to prevent the potential threats posed by cancer. In this study, the anti-tumor efficacy of attenuated Salmonella Typhimurium with Pseudomonas Exotoxin (PE38) was evaluated both macroscopically and microscopically on human breast adenocarcinoma (MCF-7) and the mechanism of the applied immunotherapeutic treatment was investigated by demonstrating the presence and pathways of apoptosis pyroptosis. In this presented study, after in vitro passage of the MCF-7 cell line, implantation was performed in Atimic Nude mice. Tumor formation and diameter were regularly monitored, and when each reached a diameter above 200 mm3, immunotherapy with avirulent Salmonella Typhimurium containing Pseudomonas exotoxin (PE38) and TGF? and PE38 (?ppGpp; relA, spoT, and Glm mutant) was applied. After treatment, systemic necropsies of euthanized mice were performed, and the tumor tissues were fixed for histopathological and immunohistochemical examinations. Histopathologically, while only adenocarcinoma-associated tumoral cells were observed in the control group mice; bacterial clusters accompanying neoplastic epithelial cells, necrotic and apoptotic figures, and inflammatory cell infiltration were found in the tumor tissues of treated mice. While intense anti-caspase 3 and 11 activities were detected in the treatment group, moderate anti-caspase 1, 8, and 9 activities, as well as mild anti-Gasdermin E immunoreactivity, were observed. In contrast, no immunoreaction was found in the control group. With this presented study, the anti-tumoral activity of avirulent Salmonella Typhimurium has been demonstrated. In this anti-tumoral mechanism, the presence of necrosis, apoptosis, and pyroptosis has been shown as the mechanisms of cell death for tumor cells. Accordingly, this is of fundamental nature for the development of a successful alternative method in breast cancer treatment and for establishing the underlying mechanisms that will serve as a basis for other cancer treatments. Keywords: Anti-tumoral activity, Apoptosis, Immunohistochemistry, MCF-7, Pyroptosis, Salmonella Typhimurium.
Anti-cancer treatments, especially current breast cancer therapies, encompass critical side effects such as appetite loss, resistance to infections, weakness and fatigue, weight gain, early menopause, and development of resistance to tumors. Therefore, there is a need for new approaches in diagnosis and treatment to prevent the potential threats posed by cancer. In this study, the anti-tumor efficacy of attenuated Salmonella Typhimurium with Pseudomonas Exotoxin (PE38) was evaluated both macroscopically and microscopically on human breast adenocarcinoma (MCF-7) and the mechanism of the applied immunotherapeutic treatment was investigated by demonstrating the presence and pathways of apoptosis pyroptosis. In this presented study, after in vitro passage of the MCF-7 cell line, implantation was performed in Atimic Nude mice. Tumor formation and diameter were regularly monitored, and when each reached a diameter above 200 mm3, immunotherapy with avirulent Salmonella Typhimurium containing Pseudomonas exotoxin (PE38) and TGF? and PE38 (?ppGpp; relA, spoT, and Glm mutant) was applied. After treatment, systemic necropsies of euthanized mice were performed, and the tumor tissues were fixed for histopathological and immunohistochemical examinations. Histopathologically, while only adenocarcinoma-associated tumoral cells were observed in the control group mice; bacterial clusters accompanying neoplastic epithelial cells, necrotic and apoptotic figures, and inflammatory cell infiltration were found in the tumor tissues of treated mice. While intense anti-caspase 3 and 11 activities were detected in the treatment group, moderate anti-caspase 1, 8, and 9 activities, as well as mild anti-Gasdermin E immunoreactivity, were observed. In contrast, no immunoreaction was found in the control group. With this presented study, the anti-tumoral activity of avirulent Salmonella Typhimurium has been demonstrated. In this anti-tumoral mechanism, the presence of necrosis, apoptosis, and pyroptosis has been shown as the mechanisms of cell death for tumor cells. Accordingly, this is of fundamental nature for the development of a successful alternative method in breast cancer treatment and for establishing the underlying mechanisms that will serve as a basis for other cancer treatments. Keywords: Anti-tumoral activity, Apoptosis, Immunohistochemistry, MCF-7, Pyroptosis, Salmonella Typhimurium.
Açıklama
Sağlık Bilimleri Enstitüsü, Veterinerlik Patolojisi Ana Bilim Dalı, Veteriner Bilim Dalı
Anahtar Kelimeler
Patoloji, Pathology
