İnsan amniyotik membran türevli medyumun, pankreas kanser hücreleri üzerinde anti-tümöral özelliklerinin araştırılması
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Tarih
2024
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Yayıncı
Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amniyotik membran (AM); embriyoyu çevreleyen, içi amniyotik sıvı ile dolu ve plasentanın iç tarafında bulunan ince bir membrandır. Plasentaya esneklik katmasıyla birlikte ona bir yaşam alanı sağlar. Son yıllarda amniyotik membran yapı ve fonksiyonları, tıbbın pek çok alanında araştırma konusu haline gelmiştir. AM; anti-viral, anti-tümöral, anti-inflamatuar ve anti-bakteriyel özelliklerin yanı sıra anti-anjiyogenik ve anti-proleratif özellikler sunmaktadır ve tıpta kullanımında etik açıdan sorun teşkil etmemektedir. Umut vaat eden fizyolojik özellikleri nedeniyle AM, artık günümüzde ölüm nedenleri arasında ilk sıralarda gösterilen kanser tedavisi için de mercek altına alınmıştır. Kadın ver erkeklerde yüksek sayılarda seyreden bir kanser türü olan pankreas kanseri için de alternatif tedavi yollarına ihtiyaç duyulmaktadır. Bu tez çalışması ile ölüm oranı giderek artmakta olan pankreas kanseri üzerinde amniyotik membran ile koşullandırılmış medyumun etkilerinin araştırılması hedeflenmiştir. Aynı zamanda AM kullanılarak ilk kez pankreas kanseri hücrelerine yönelik bir çalışma yapılmıştır. AM kullanılarak oluşturulan kondisyon medyumunun etkileri proliferasyon, migrasyon, invazyon, gen ekspresyonu ve protein bazında incelenmiştir. Proliferasyon için XTT analizi yapıldı ve hAM-CM ile muamele edilen PANC-1 hücrelerinin canlılığında anlamlı bir azalma olduğu görülmüştür. hAM-CM ile muamele edilen HEK-293 hücrelerinin morfolojilerinde herhangi bir değişiklik gözlenmezken, PANC-1 hücrelerinin morfolojisinde anlamlı değişiklikler gözlenmiştir. Migrasyon analizi için yara tahlili deneyi kullanıldı ve hAM-CM'nin HEK-293 hücrelerinin migrasyonuna anlamlı bir etkisi olmadığı (p˃0,05) görüldü. Buna karşılık PANC-1 hücrelerinin migrasyonunu anlamlı seviyede inhibe ettiği (p<0,05) görülmüş olup DMEM ve hAM-CM ile muamele edilen grupların yara kapanma yüzdeleri sırasıyla %89 ve %11,83 olarak hesaplandı. İnvazyon yüzdelerine bakıldığında ise HEK-293 hücrelerinin invazyon yüzdeleri birbirine yakın (%37,93 ve % 36,88) hesaplanırken PANC-1 hücrelerinin invazyon yüzdeleri DMEM ve hAM-CM ile muamele edilen grupta sırasıyla %23,70 ve %13,15 olarak hesaplandı. Gen ekspresyon sonuçlarına göre ise hAM-CM, PANC-1 hücrelerinde 48 saatlik muameleden sonra IL1β, IL-6, IL-17 ve HIF1α genlerinin ifadesini artırmıştır (p ˃ 0,06, p<0,06, p ˃ 0,03, p<0,01). ELISA sonuçlarında 48 saat hAM-CM ile muamele edilen PANC-1 hücrelerinin kontrole göre total IL-17 proteinin miktarını azaltırken, total HIF1α ve total VEGF proteinin miktarını arttırdığı gözlenmiştir. Bununla birlikte, IL-17 sekrete proteininin miktarını arttırmakta ve HIF1α ve VEGF sekrete proteinlerinin miktarını azaltmaktadır. Sonuç olarak hAM-CM muamelesi; PANC-1 hücrelerinin morfolojisinde değişikliğe yol açarak, hücre canlılığı, migrasyon ve invazyon yeteneğini düşürmede etkili olmuştur. hAM-CM'nin anti-tümöral özelliklerinin daha fazla çalışmayla aydınlatılmasına ihtiyaç vardır.
The amniotic membrane (AM) is a thin membrane surrounding the embryo, filled with amniotic fluid and located on the inner side of the placenta. Along with adding flexibility to the placenta, it provides it with a living space. In recent years, the structure and functions of the amniotic membrane have become the subject of research in many fields of medicine. AM offers anti-viral, anti-tumoral, anti-inflammatory and anti-bacterial properties as well as anti-angiogenic and anti-prolerative properties and does not pose ethical problems in its use in medicine. Due to its promising physiological properties, AM has also been put under the spotlight for the treatment of cancer, which is now ranked first among the causes of death today. Dec. Alternative treatment options are also needed for pancreatic cancer, a type of cancer that occurs in high numbers in women and men. This thesis study aims to investigate the effects of medium conditioned with amniotic membrane on pancreatic cancer, whose mortality rate is increasing. At the same time, a study was conducted on pancreatic cancer cells for the first time using AM. The effects of the conditioning medium created using AM have been studied on the basis of proliferation, migration, invasion, gene expression and protein. XTT analysis was performed for proliferation and it was found that there was a significant decrease in the viability of PANC-1 cells treated with hAM-CM. While no changes were observed in the morphologies of HEK-293 cells treated with hAM-CM, significant changes were observed in the morphology of PANC-1 cells. A wound assay experiment was used for migration analysis, and it was found that HAM-CM did not have a significant effect on the migration of HEK-293 cells (p0,05). In contrast, it was found to significantly inhibit the migration of PANC-1 cells (p<0.05), and the wound closure percentages of the groups treated with DMEM and hAM-CM were calculated as 89% and 11.83%, respectively. Looking at the invasion percentages, the invasion percentages of HEK-293 cells were calculated close to each other (37.93% and 36.88%), while the invasion percentages of PANC-1 cells were calculated as 23.70% and 13.15% in the group treated with DMEM and hAM-CM, respectively. According to the gene expression results, hAM-CM increased the expression of IL1ß, IL-6, IL-17 and HIF1a genes in PANC-1 cells after 48 hours of treatment (p 0.06, p<0.06, p 0.03, p<0.01). ELISA results showed that PANC-1 cells treated with hAM-CM for 48 hours decreased the amount of total IL-17 protein and increased the amount of total HIF1α and total VEGF protein compared to the control. However, it increases the amount of IL-17 secreted protein and decreases the amount of HIF1α and VEGF secreted proteins. As a result, hAM-CM treatment has been effective in reducing cell viability, migration and invasion ability by causing changes in the morphology of PANC-1 cells. The anti-tumoral properties of HAM-CM need to be elucidated by further studies.
The amniotic membrane (AM) is a thin membrane surrounding the embryo, filled with amniotic fluid and located on the inner side of the placenta. Along with adding flexibility to the placenta, it provides it with a living space. In recent years, the structure and functions of the amniotic membrane have become the subject of research in many fields of medicine. AM offers anti-viral, anti-tumoral, anti-inflammatory and anti-bacterial properties as well as anti-angiogenic and anti-prolerative properties and does not pose ethical problems in its use in medicine. Due to its promising physiological properties, AM has also been put under the spotlight for the treatment of cancer, which is now ranked first among the causes of death today. Dec. Alternative treatment options are also needed for pancreatic cancer, a type of cancer that occurs in high numbers in women and men. This thesis study aims to investigate the effects of medium conditioned with amniotic membrane on pancreatic cancer, whose mortality rate is increasing. At the same time, a study was conducted on pancreatic cancer cells for the first time using AM. The effects of the conditioning medium created using AM have been studied on the basis of proliferation, migration, invasion, gene expression and protein. XTT analysis was performed for proliferation and it was found that there was a significant decrease in the viability of PANC-1 cells treated with hAM-CM. While no changes were observed in the morphologies of HEK-293 cells treated with hAM-CM, significant changes were observed in the morphology of PANC-1 cells. A wound assay experiment was used for migration analysis, and it was found that HAM-CM did not have a significant effect on the migration of HEK-293 cells (p0,05). In contrast, it was found to significantly inhibit the migration of PANC-1 cells (p<0.05), and the wound closure percentages of the groups treated with DMEM and hAM-CM were calculated as 89% and 11.83%, respectively. Looking at the invasion percentages, the invasion percentages of HEK-293 cells were calculated close to each other (37.93% and 36.88%), while the invasion percentages of PANC-1 cells were calculated as 23.70% and 13.15% in the group treated with DMEM and hAM-CM, respectively. According to the gene expression results, hAM-CM increased the expression of IL1ß, IL-6, IL-17 and HIF1a genes in PANC-1 cells after 48 hours of treatment (p 0.06, p<0.06, p 0.03, p<0.01). ELISA results showed that PANC-1 cells treated with hAM-CM for 48 hours decreased the amount of total IL-17 protein and increased the amount of total HIF1α and total VEGF protein compared to the control. However, it increases the amount of IL-17 secreted protein and decreases the amount of HIF1α and VEGF secreted proteins. As a result, hAM-CM treatment has been effective in reducing cell viability, migration and invasion ability by causing changes in the morphology of PANC-1 cells. The anti-tumoral properties of HAM-CM need to be elucidated by further studies.
Açıklama
Anahtar Kelimeler
Migrasyon, Migration, Amniyotik Membran, Gen Ekspresyonu, Kondisyon Medyumu, Pankreas Kanseri, Pancreatic Cancer, Conditioning Medium, Gene Expression, Amniotic Membrane
Kaynak
WoS Q Değeri
Scopus Q Değeri
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Sayı
Künye
Ertuğrul, A. (2024). İnsan amniyotik membran türevli medyumun, pankreas kanser hücreleri üzerinde anti-tümöral özelliklerinin araştırılması. (Yüksek Lisans Tezi). Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü, Konya.
