İnsan diş pulpası kaynaklı mezenkimal kök hücrelerin meme kanseri kök hücreleri üzerine etkisi

Gizem İnetaş Yengin; Öykü Gönül Geyik; Hüsnü Alper Bağrıyanık; Zeynep Yüce; Gülperi Öktem

doi:10.19161/etd.1360783

TR EN

İnsan diş pulpası kaynaklı mezenkimal kök hücrelerin meme kanseri kök hücreleri üzerine etkisi

Öz

Amaç: Kanser kök hücreleri; kendini yenileyebilen, farklılaşma kapasitesi yüksek ve uzun süreli proliferasyon ile normal dokuya invazyon kabiliyeti olan hücrelerdir. Bu yetenekleriyle geleneksel kanser tedavisine direnç oluşturarak tümör büyümesi ve metastazda rol oynar. Başarılı kanser tedavileri için kanser kök hücre mekanizmalarına yönelik araştırmalar yapmak önem taşımaktadır. Bu çalışmanın amacı, insan diş pulpası kaynaklı mezenkimal kök hücrelerin meme kanseri kök hücreleri üzerine etkisinin hücre canlılığı, hücre döngüsü ve apoptoz yöntemleriyle araştırılmasıdır. Gereç ve Yöntem: Meme kanseri hücreleri (MCF7) akış sitometrisi ile CD44+/CD24- boyaması yapılarak ayrılmıştır. CD44+/CD24- popülasyonuna meme kanseri kök hücresi denilmiştir. Diş pulpasından izole edilen mezenkimal kök hücreler kültüre edilip karakterizasyonu yapılmıştır. Mezenkimal kök hücre grubu mCitrine, meme kanseri kök hücresi grubu ise mCherry ile plazmit transfeksiyonu yapılarak işaretlenmiştir. Bu hücreler 48 saat boyunca ko-kültüre edilmiş ve sonrasında hücre canlılığı, hücre döngüsü ve apoptoz analizleri yapılmıştır. Bulgular: Diş pulpası kaynaklı mezenkimal kök hücreler ile ko-kültüre edilen meme kanseri kök hücrelerinin kontrol grubuna göre hücre canlılığı, hücre döngüsü ve apoptoz değerlerinde zamana bağlı olarak istatistiksel anlamlı değişiklikler görülmüştür. Ko-kültüre grubu kontrole göre kıyaslandığında zamana bağlı olarak G0/G1 evresinde artış gözlenmiştir. Ko-kültüre edilen hücrelerin floresan mikroskop ile yapılan incelemesinde sarı floresan işaretli hibrit hücreler gözlenmiştir ve immüno-floresan Ki67 boyamasında hücre sayısında azalma gözlenmiştir. Sonuç: Ko-kültür sonrası diş pulpası kaynaklı mezenkimal kök hücrelerin meme kanser kök hücreleri üzerinde hücre proliferasyonunu inhibe edici etkileri olduğu ve apoptozu teşvik ettiği gözlenmiştir. Sonuç olarak, meme kanser kök hücreleri üzerinde diş pulpası kaynaklı mezenkimal kök hücrelerin tedaviye yönelik bir etkisi olabilir.

Anahtar Kelimeler

Effect of human dental pulp derived mesenchymal stem cells on breast cancer stem cells

Abstract

Aim: Cancer stem cells are cells that can renew themselves, have a high differentiation capacity and have the ability to invade normal tissue with long-term proliferation. With these abilities, it plays a role in tumor growth and metastasis by creating resistance to traditional cancer treatment. It is important to conduct research on cancer stem cell mechanisms for successful cancer treatments. The aim of this study is to investigate the effects of human dental pulp-derived mesenchymal stem cells on breast cancer stem cells by cell viability, cell cycle and apoptosis methods. Materials and Methods: Breast cancer cells (MCF7) were separated by flow cytometry with CD44+/CD24- staining. The CD44+/CD24- population was called breast cancer stem cells. Mesenchymal stem cells isolated from dental pulp were cultured and characterized. Mesenchymal stem cell group was labeled with mCitrine and breast cancer stem cell group with mCherry by plasmid transfection. These cells were co-cultured for 48 hours and then cell proliferation, cell cycle and apoptosis analyzes were performed. Results: Statistically significant changes were observed in cell viability, cell cycle and apoptosis values of breast cancer stem cells co-cultured with dental pulp-derived mesenchymal stem cells compared to the control group over time. When the co-culture group was compared to the control, an increase in the G0/G1 stage was observed depending on time. Yellow fluorescent-labeled hybrid cells were observed in the examination of co-cultured cells with fluorescent microscope, and a decrease in cell number was observed in immunofluorescent Ki67 staining. Conclusion: When mesenchymal stem cells from dental pulp were co-cultured with breast cancer stem cells, an increase in the G0/G1 stage of breast cancer stem cells was observed depending on time. After co-culture, it has been observed that dental pulp-derived mesenchymal stem cells have cell proliferation inhibitory effects on breast cancer stem cells and promote apoptosis. In conclusion, dental pulp derived mesenchymal stem cells may have a therapeutic effect on breast cancer stem cells.

Keywords

Thanks

Çalışma, Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi TYL-2019-20624 numaralı proje kapsamında desteklenmiştir. Çalışmada dental pulpa temini katkılarından dolayı Dr. Dt. İlkay Aydoğdu ve Doç.Dr. Onur Şahin’e teşekkür ederiz.

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Details

Primary Language

Turkish

Subjects

Regenerative Medicine (Incl. Stem Cells) , Histology and Embryology

Journal Section

Research Article

Authors

Gizem İnetaş Yengin ^*
0000-0002-9421-651X
Türkiye

Öykü Gönül Geyik
0000-0003-3014-1253
Türkiye

Hüsnü Alper Bağrıyanık
0000-0003-4064-7637
Türkiye

Zeynep Yüce
0000-0002-2762-0942
Türkiye

Gülperi Öktem
0000-0003-1914-2503
Türkiye

Publication Date

September 18, 2023

Submission Date

December 6, 2022

Acceptance Date

March 9, 2023

Published in Issue

Year 1970 Volume: 62 Number: 3

DOI

https://doi.org/10.19161/etd.1360783

IZ

https://izlik.org/JA44RX65XU

Cite

RIS / Bibtex

Vancouver

1.Gizem İnetaş Yengin, Öykü Gönül Geyik, Hüsnü Alper Bağrıyanık, Zeynep Yüce, Gülperi Öktem. İnsan diş pulpası kaynaklı mezenkimal kök hücrelerin meme kanseri kök hücreleri üzerine etkisi. EJM. 2023 Sep. 1;62(3):396-405. doi:10.19161/etd.1360783