YUMURTALIK KANSERİ HÜCRELERİ İLE BİRLİKTE KÜLTÜRLENEN MEME HÜCRELERİ ÜZERİNDE DNAZ I ETKİSİ: HÜCRESEL ETKİLEŞİMLER VE CANLILIK ÜZERİNE BİR ARAŞTIRMA

Year 2024, Volume: 25 Issue: 4, 351 - 364, 22.12.2024

Didem Turgut Coşan , İbrahim Uğur Çalış

https://doi.org/10.69601/meandrosmdj.1540484

Abstract

Giriş: Hücre dışı DNAlar (cf-DNA'lar), hücre apoptozu, nekroz veya yaşayan hücrelerden aktif salgılama gibi mekanizmalarla kan dolaşımına girebilir ve kanser hücrelerinin plazma ve seruma saldığı DNA'lar da bunların kaynağı olabilir. Çeşitli çalışmalar, cf-DNA'ların kanser gelişimi ve metastazdaki önemli rolünü vurgulamıştır. Bu cf-DNA'lar, hücre hasarı, apoptoz veya aktif salgılama nedeniyle kan dolaşımına girebildiği gibi, kanser hücreleri tarafından da salınabilir. Bilim insanları arasında cf-DNA'ların kanserin ilerlemesi ve metastaz üzerindeki etkileri konusunda süregelen bir tartışma vardır. Bu çalışma, yumurtalık kanseri hücrelerinin tümörojenik olmayan meme hücrelerinin çoğalma ve yaşama yeteneklerini nasıl etkilediğini araştırmayı ve ilk gözlemleri sunmayı amaçlamaktadır.
Yöntem ve Gereçler: İnsan yumurtalık kanseri hücreleri ve normal insan meme hücreleri, laboratuvarımızda uygun ortamlarda %5 CO2 içeren 37°C'de kültüre edilmiştir. Ortak kültür deneyleri için hücreler, DNase I içeren veya içermeyen ortamlarda 72 saat boyunca plakalar ve insertlerde ayrı ayrı inkübe edilmiştir. Hücre canlılığı, tripan mavisi dışlama testi ile değerlendirilirken, çoğalma ve yapışma XTT test kiti ile analiz edilmiştir.
Sonuçlar: Bu çalışma, OVCAR-3 hücreleri ile birlikte kültüre edilen MCF-10A hücrelerinin canlılık ve çoğalma oranlarının arttığını gözlemlemiştir. Ortak kültürdeki çoğalma, tekil kültürlerle kıyaslandığında daha yüksekti. DNase I uygulaması, OVCAR-3 hücrelerinin çoğalma (p<0.01) ve canlılığını önemli ölçüde azaltmış, ancak MCF-10A hücrelerinin çoğalma ve canlılığı üzerinde anlamlı bir etki göstermemiştir. DNase I varlığında veya yokluğunda OVCAR-3 hücrelerine özgü p53 ekzon 7 mutasyonları veya hiper-metilasyon tespit edilmemiştir.

Keywords

cf-DNA, OVCAR-3, MCF-10A, Kanser, DNaz I

Project Number

201811066

DNASE I IMPACT ON BREAST CELLS CO-CULTURED WITH OVARIAN CANCER CELLS: A STUDY OF CELLULAR INTERACTIONS AND VIABILITY

Abstract

Introduction: Cell-free DNAs (cf-DNAs) are released into the bloodstream through cell apoptosis, necrosis, or active secretion, often originating from cancer cells. These cf-DNAs have been associated with cancer development and metastasis, although their precise role remains under debate. DNase I, an enzyme that degrades extracellular DNA, has shown potential to impact cf-DNAs and influence cancer progression. This study investigates the effects of ovarian cancer cells on the proliferation and viability of non-tumorigenic breast cells, with a focus on DNase I’s role.
Materials and Methods: Human ovarian cancer cells (OVCAR-3) and normal human breast cells (MCF-10A) were cultured under standard conditions (37°C, 5% CO₂). Co-culture experiments were conducted by incubating cells separately in plates and inserts, with or without DNase I, for 72 hours. Cell viability was assessed using the trypan blue exclusion test, while proliferation and adhesion were measured with an XTT assay.
Results: DNase I significantly reduced OVCAR-3 proliferation (p<0.001) and adhesion (p<0.01) without affecting MCF-10A cells. DNase I also decreased OVCAR-3 cell viability but did not significantly impact MCF-10A viability. Genetic analysis identified p53 exon 7 mutations and methylation of APC1A, APC1B, and RASSF1A genes in OVCAR-3 cells, which were unaffected by DNase I. No mutations or methylation were detected in MCF-10A cells.
Conclusion: The results suggest that the impact of DNase I on the proliferation and viability of cancer cells is significant and warrants further investigation. The potential effects of prolonged exposure between different cell types could yield even more compelling findings.

Keywords

cf-DNA, OVCAR-3, MCF-10A, cancer, methylation, DNase I

Ethical Statement

Çalışma ticari olarak satın alınan hücre kültürleri üzerinde yapıldığından etik kurul onay belgesine gerek yoktur.

Supporting Institution

This study was supported by a Grant from the Research Foundation of Eskisehir Osmangazi University, Türkiye (Project no: 201811066).

Project Number

201811066

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