Effect of cisplatin and mitochondria transplantation isolated from human mesenchymal stem cells on DU-145 cell proliferation

Year 2021, , 1285 - 1292, 30.09.2021

Zehra Çiçek , Volkan Tekin

https://doi.org/10.17826/cumj.912336

Abstract

Purpose: The aim of this study is to evaluate proliferation and cytotoxicity by applying the antineoplastic drug cisplatin and mitochondria isolated from human mesenchymal stem cells (IMSCs) to human prostate cancer cells (DU-145).
Materials and Methods: IMSCs and DU-145 cell line were used in our study. IMSCs were multiplied to ~20x106 cells and mitochondria were isolated according to the protocols in kit. Protein content of the isolated mitochondria was measured by bicinchoninic acid method (BCA). DU-145 cells were seeded into 96-well plates at 10x103 cells in per well. Cisplatin 6 µM and 14 µM doses were administered for 24 hours. Isolated mitochondria (10x, 100x) were incubated for 24 hours again. Cell proliferation and viability were measured using the tetrazolium salt (MTT) method, and the absorbance values of 570 nm-630 nm were measured in a plate reader, and the cell proliferation percentages were calculated.
Results: 6 µM cisplatin application to DU-145 cells proliferation decreased by 35% compared to the control group, while 14 µM cisplatin reduced it by 68%. When compared with the Cis-6 µM group, there was no significant difference in proliferation in the Cis-6 µM+Mito-10x group while a significant decrease was observed in the Cis-6 µM+Mito-100x group.
Conclusion: When mitochondria are transplanted together with cisplatin to prostate cancer cells, proliferation is further reduced. In addition, mitochondria transplantation may reduce the proliferation of cancerous cells by affecting various intracellular proliferative, apoptotic signaling pathways and by regulating the microenvironment.

Keywords

Stem cell, culture, mitochondria transplantation

Sisplatin ve insan mezenkimal kök hücrelerden izole edilen mitokondri naklinin DU-145 hücre proliferasyonuna etkisi

Abstract

Amaç: Bu çalışmada antineoplastik bir ilaç olan sisplatin ve insan mezenkimal kök hücrelerinden (İMKH) izole edilen mitokondriler, insan prostat kanser hücrelerine (DU-145) uygulanarak, proliferasyon ve sitotoksisitenin değerlendirilmesi hedeflenmiştir.
Gereç ve Yöntem: Çalışmamızda, İMKH ve DU-145 hücre hatları kullanıldı. İMKH’ ler ~20x106 hücre olacak şekilde çoğaltılarak kitte belirtilen protokollere göre mitokondriler izole edildi. İzole mitokondrilerin protein miktarı, bikinkoninik asit yöntemiyle (BCA) ölçüldü. DU-145 hücreleri, her kuyuda 10x103 hücre olacak şekilde 96’ lık plate’e ekildi. Sisplatin’ in 6 µM ve 14 µM dozları 24 saat uygulandı. İnkübasyon sonunda hücre medyumları değiştirildi ve izole mitokondriler (10x, 100x) eklenerek tekrar 24 saat inkübe edildi. Hücre çoğalması ve canlılığı tetrazolyum tuzu (MTT) yöntemiyle, 570nm-630nm absorbans değerleri plate okuyucu da ölçülerek farkları alındı ve hücre çoğalma yüzdeleri hesaplandı.
Bulgular: DU-145 hücrelerine 6 µM sisplatin uygulaması kontrol grubuna göre proliferasyonu %35 oranında azaltırken, 14 µM sisplatin ise hücre proliferasyonunu %68 azalttı. Cis-6 µM grubuyla karşılaştırıldığında, Cis-6 µM+Mito-10x grubunda proliferasyonda anlamlı fark görülmezken, Cis-6 µM+Mito-100x grubunda anlamlı azalma görüldü.
Sonuç: Prostat kanseri hücrelerine sisplatin ile birlikte mitokondri transplantasyonu yapıldığında proliferasyonu daha da azalmaktadır. Ayrıca, mitokondri transplantasyonunun intrasellüler çeşitli proliferatif, apopitotik sinyal yolaklarını etkileyerek ve mikroçevreyi düzenleyerek kanserli hücrelerin çoğalmasını azaltabilir.

Keywords

kök hücre, kültür, mitokondri

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