Hidrojen peroksit ve mitokondrinin insan meme kanseri hücrelerine etkisi

Year 2023, Volume: 48 Issue: 1, 109 - 116, 31.03.2023

Zehra Çiçek

https://doi.org/10.17826/cumj.1187258

Abstract

Amaç: Bu çalışmada, hidrojen peroksit (H2O2), insan mezenkimal kök hücreleri (İMKH) ve insan meme kanseri hücrelerinden (MCF-7) izole edilen mitokondrilerin, MCF-7 hücre proliferasyonuna etkisinin değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntem: İMKH ve MCF-7 hücrelerinin mitokondrileri kitte belirtilen protokollere göre izole edildi ve protein miktarı bikinkoninik asit yöntemiyle (BCA) ölçüldü. H2O2 dozları (0-1000 µM), MCF-7 ve İMKH’lerden elde edilen mitokondrilerin, MCF-7 hücrelerine transferi sonrasında hücre canlılığı tetrazolyum tuzu (MTT) yöntemiyle değerlendirildi. Örneklerin absorbans değerleri (570 nm) plate okuyucuyla ölçülerek hücre yüzdeleri hesaplandı.
Bulgular: İMKH’lerden izole edilen mitokondrilerin ATP düzeyinin, MCF-7’lerden daha yüksek olduğu bulundu. MCF-7 hücrelerinin canlılığı H2O2’nin 30 µM (%19), 40 µM (%26), 50 µM (%39), 75 µM (%39), 100 µM (%36), 200 µM (%28), 400 µM (%23), 800 µM (%22) ve 1000 µM dozunda %23 oranında anlamlı olarak azaldı. İMKH’lerin mitokondrilerinin tek başına transferinin hücre proliferasyonunda azalma eğilimine neden olduğu belirlenirken, herhangi bir anlamlılık tespit edilmedi. Ancak, 10 µM ve 1000 µM H2O2’yle birlikte yapılan sağlıklı mitokondri transferinin MCF-7’lerin çoğalmasını azalttığı belirlendi (sırasıyla %14 ve %8).
Sonuç: H2O2 ve İMKH’lerden izole edilen sağlıklı mitokondrilerin MCF-7 hücrelerine transferinin proliferatif süreçlerle ilişkili olduğu görülürken, kanser hücrelerine sağlıklı mitokondri naklinin, hastalığın tedavisinde umut vadeden yeni bir yöntem olabileceği düşünülmektedir.

Keywords

kök hücre, kültür, proliferasyon

The effects of hydrogen peroxide and mitochondria on human breast cancer cells

Abstract

Purpose: It was aimed to evaluate the effect of hydrogen peroxide (H2O2), mitochondria isolated from human breast cancer cells (MCF-7) and human mesenchymal stem cells (IMSCs) on MCF-7 cell proliferation in this study.
Materials and Methods: Mitochondria of IMSCs and MCF-7 cells were isolated according to the protocols specified in the kit and the protein amount of mitochondria was measured by the bicinchoninic acid method (BCA). H2O2 doses (0-1000 µM), mitochondria obtained from MCF-7 and IMSCs were transferred to MCF-7 cells and the viability was evaluated by tetrazolium salt (MTT) method. The cell percentages were calculated by measuring absorbance of the samples at 570 nm with a plate reader.
Results: It was found that, ATP level of the mitochondria isolated from IMSCs was higher than MCF-7s. MCF-7 cell viability decreased significantly in 30 µM (%19), 40 µM (%26), 50 µM (%39), 75 µM (%39), 100 µM (%36), 200 µM (%28), 400 µM (%23), 800 µM (%22) and 1000 µM (%23) H2O2 doses. The transfer of IMSCs mitochondria caused a tendency to decrease in cell proliferation, but no significance was detected. But, it was found that healthy mitochondria transfer with 10 µM and 1000 µM H2O2 reduced the proliferation of MCF-7s (respectively %14 and % 8).
Conclusion: It was determined that the transfer of healthy mitochondria isolated from IMSCs and H2O2 to MCF-7 is associated with proliferative processes, however transplantation of the healthy mitochondria into cancer cells is thought to be a promising new method in the treatment of the disease.

Keywords

stem cell, culture, proliferation

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