Wharton Jölesi Kökenli Mezenkimal Kök Hücrelerden Elde Edilen Eksozomların Akciğer, Pankreas ve Karaciğer Kanseri Hücrelerinin Canlılığı ve Metabolik Aktivitesi Üzerindeki Etkilerinin Araştırılması

Abstract

Amaç: Mezenkimal kök hücreler (MKH) sitokinler, kemokinler, eksozomlar salgılayarak veya doğrudan temas yoluyla aynı mikroçevredeki kanser hücreleri üzerinde etki gösterir. WJ-MKH kökenli eksozomların akciğer, pankreas ve karaciğer kanseri hücre hatlarının canlılığı ve metabolik aktivitesi üzerindeki etkileri incelenmesi amaçlanmıştır. Yöntem: Bu çalışmada Wharton Jölesi (WJ)-MKH kökenli eksozomlar A549, Panc-1 ve HepG2 hücrelerine 1-5-10-15 ve 20 milyon partikül eksozom olarak uygulandı. Canlılık üzerindeki etkiler 48 ve 96. saatlerde MTT analizi ile değerlendirilmiştir. Metabolik aktivitesi üzerindeki etkiler 96. saatin sonunda kalsein boyaması ile floresan yoğunluğu ile değerlendirilmiştir. Hücre metabolik aktivitesi floresan yoğunluğu kullanılarak hesaplanmıştır. Bulgular: Kontrol grubunda 96. saatin sonunda A549 hücrelerinde % 62, Panc-1 hücrelerinde %95 ve HepG2 hücrelerinde %67 artış gözlemlenirken 20 milyon eksozom uygulanan grupta ise A549 hücrelerinde %36, Panc-1 hücrelerinde %43 ve HepG2 hücrelerinde %31 artış gözlenmiştir. Proliferasyon oranı 20 milyon eksozom uygulanan grupta kontrol grubuna göre (% 100) A549: %61, Panc-1: %64 ve HepG2: %68 olarak hesaplanmıştır. Ön bulgular, WJ-MSC kökenli eksozomların kanser hücrelerine uygulandığında kanser hücrelerinin canlılığını ve metabolik aktivitesini artırmadığını ve zaman-konsantrasyona bağlı olarak azalabildiği de gösterilmiştir. Sonuç: WJ-MKH türevi eksozomlar, yüksek stroma içeriğine sahip tümörlerde in vitro hücre canlılığı ve metabolik aktivite üzerinde etkili olabilir ve tümör hücreleri üzerindeki diğer mekanizmalar üzerindeki etkilerinin araştırılması gerekmektedir.

Keywords

• WJ-MKH
• MKH kökenli eksozom
• kanser hücresi
• tümör stroması

Exploring the Effects of WJ-MSC-Derived Exosomes on the Viability and Metabolic Activity of Lung, Pancreatic and Liver Cancer Cells

Abstract

Aim: Mesenchymal stem cells (MSCs) act on cancer cells in the same microenvironment by secreting cytokines, chemokines, exosomes, or by direct contact. The purpose of this study is to analyse the impact of WJ-MSC-derived exosomes on the viability and metabolic activity of lung, pancreatic, and liver cancer cells. Method: In this study, Wharton's Jelly (WJ)-MSCs-derived exosomes were added to A549, Panc-1, and HepG2 cells as 1-5-10-15 and 20 million particle exosomes. The effects on viability were evaluated by MTT analysis at 48 and 96 hours. The effects on metabolic activity were evaluated by calcein staining at the end of the 96th hour. Cell metabolic activity was calculated using fluorescence intensity. Results: At the end of the 96th hour, 62% increase in A549 cells, 95% increase in Panc-1 cells and 67% increase in HepG2 cells were detected in the control group. In the group which 20 million exosomes were applied, 36% increase in A549 cells, 43% increase in Panc-1 cells and 31% increase in HepG2 cells were observed. The fluorescence intensity of 20 million exosomes was calculated as: A549: 61%, Panc-1: 64% and HepG2: 68%. Preliminary findings indicate that WJ-MSC-derived exosomes do not increase the viability and metabolic activity of cancer cells when applied to them. It was also shown that the viability and metabolic activity can decrease depending on time and concentration. Conclusion: WJ-MSC-derived exosomes may be effective on cell viability and metabolic activity in vitro in tumors with high stroma content, and their effects on other mechanisms on tumor cells need to be investigated.

Keywords

• WJ-MSC
• MSC derived exosome
• cancer cell
• tumor stroma

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