KOLAJENAZ UYGULAMA SÜRESİNİN PRİMER MEME KANSERİ HÜCRE PROTEOMUNA ETKİLERİ

Year 2024, , 222 - 232, 30.09.2024

Merve Gulsen Bal Albayrak , Murat Kasap , Gürler Akpınar , Sevinc Yanar , Turgay Şimşek , Zafer Cantürk

https://doi.org/10.69601/meandrosmdj.1542417

Abstract

Amaç: Hücrelerin davranış mekanizmaları ve hastalıkların mekanizmalarını incelemek için kritik öneme sahip olan primer hücre izolasyonu metodunda kollajenaz aracılı doku ayrıştırması kritik bir rol oynamaktadır. Ancak, kollajenaz enziminin uygulama süresinin primer hücrelerin proteomuna, özellikle de meme kanseri araştırmalarında, etkisi yeterince araştırılmamıştır. Bu çalışma, kollajenaz II uygulama süresinin birincil meme kanseri hücrelerinin proteomik profilleri üzerindeki etkilerini araştırmayı amaçlamaktadır.
Materyal ve Metotlar: İnvaziv duktal karsinom teşhisi konmuş hastalardan alınan meme kanseri dokuları, kollajenaz II ile 1 saat ve 3 saat süreyle muamele edilmiştir. Sonrasında proteomik analiz, sıvı kromatografisi-tandem kütle spektrometrisi (LC-MS/MS) kullanılarak gerçekleştirilmiştir. Tanımlanan proteinler, farklı tedavi sürelerinin neden olduğu proteomik değişikliklerin işlevsel etkilerini belirlemek için biyoinformatik analizlere tabi tutulmuştur.
Sonuçlar: Biyoinformatik analizler, 1 saatlik uygulamanın öncelikli olarak hücre iskeleti organizasyonu ve hücre yapışmasıyla ilgili proteinleri etkilediğini ve aktin hücre iskeleti dinamikleri ile yapısal molekül aktivitelerinde belirgin zenginleşme olduğunu göstermiştir. Buna karşılık, 3 saatlik tedavi önemli metabolik yeniden programlamalara yol açmış ve enerji üretimi ile ilgili yolların, özellikle TCA döngüsü ve glikoliz, düzenlenmesini artırmıştır.
Sonuç: Bu çalışma, kollajenaz II tedavi süresinin, birincil meme kanseri hücrelerinin proteomik profilini önemli ölçüde değiştirdiğini ve daha kısa sürelerin yapısal proteinleri, daha uzun sürelerin ise metabolik değişiklikleri indüklediğini ilk kez ortaya koymaktadır. Bu sebeple, hedefli proteomik çalışmalar için tedavi süresinin optimize edilmesi oldukça önemlidir.

Keywords

Primer Hücre, meme kanseri, Kolajenaz, uygulama süresi, proteomik

DECIPHERING THE IMPACT OF COLLAGENASE TREATMENT DURATION ON PRIMARY BREAST CANCER CELL PROTEOME: A COMPREHENSIVE STUDY

Abstract

Aim: Primary cell isolation is essential for studying cellular behavior and disease mechanisms, with collagenase-mediated tissue dissociation playing a critical role in the process. However, the impact of collagenase treatment duration on the proteome of primary cells, particularly in breast cancer research, remains underexplored. This study aims to investigate the effects of collagenase II treatment duration on the proteomic profiles of primary breast cancer cells.
Materials and Methods: Breast cancer tissues from patients diagnosed with infiltrating ductal carcinoma were treated with collagenase II for either 1 or 3 hours. Subsequent proteomic analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Identified proteins were subjected to bioinformatic analyses to determine the functional implications of the proteomic changes induced by the different treatment durations.
Results: Bioinformatic analyses showed that 1-hour treatment predominantly affected proteins involved in cytoskeletal organization and cell adhesion, with significant enrichment in actin cytoskeleton dynamics and structural molecule activity. In contrast, 3-hour treatment led to significant metabolic reprogramming, with enhanced regulation of pathways involved in energy production, including the TCA cycle and glycolysis.
Conclusion: This study reveals for the first time that, collagenase II treatment duration significantly alters the proteomic profile of primary breast cancer cells, with shorter durations affecting structural proteins and longer durations inducing metabolic changes. Optimizing treatment time is crucial for targeted proteomic studies.

Keywords

Primary cells, collagenase, treatment duration, proteomics

Ethical Statement

This study has been approved by the Ethics Committee of Kocaeli University with approval number; KU GOKAEK-2019/16.04 2019/139.

Supporting Institution

Not applicable

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