Meme kanseri hücrelerinde anastaz fenomeninin tanımlanması

Amaç: Çalışmada, dünya genelinde en yaygın kanser tiplerinden biri olan meme kanserinde, apoptoz ile ilişkili genlerin anastaz sürecindeki ifadesel değişimlerinin ve potansiyel rollerinin tanımlanması amaçlanmıştır. Gereç ve Yöntem: Farklı tipteki meme kanseri hücre hatları (MCF7 ve MDA-MB-231), meme kanseri kök hücreleri ve sağlıklı meme hücre hattı (MCF10A) kullanıldı. Apoptotik ve anastatik hücre yüzdeleri, akış sitometrisi aracılığıyla Annexin V testi ile belirlendi. Apoptotik hücrelere göre anastatik hücrelerin gen ekspresyon değişimleri qRT-PCR ve 2-ΔΔCt yöntemi ile belirlendi. Anlamlı değişim gösteren genlerin görev aldıkları yolak ve biyolojik süreçler STRING v11.5 veri tabanı kullanılarak belirlendi. Bulgular: Tüm hücre gruplarında etanol uygulaması sonucu kontrole göre apoptoz yüzdesinin arttığı ve apoptozu uyaran etmenin uzaklaştırılmasıyla apoptotik hücre yüzdesinin azaldığı belirlendi. Kontrol, apoptoz ve anastaz grupları arasında apoptotik hücre yüzdesindeki değişim en fazla MCF7 hücrelerinde belirlendi. Uyumlu şekilde bu hücre hattında en fazla sayıda gende ifade değişimi belirlendi. CASP7 ve APAF1 genleri tüm hücre hatlarında ekspresyon azalışı sergiledi. Tüm hücre gruplarında anastazın sistein tipi endopeptidaz aktivitesini (GO_ID: 0043027) ve ilaç direnci ilişkili yolakları (KEGG_ID: hsa01524) ortak şekilde etkilediği belirlendi. Sonuç: Hücrelerde anastaz fenomeninin apoptoz düzenleyici mekanizmalar ile etkileşiminin tanımlanması, hem sağlıklı hücrelerin onkojenik dönüşümünün hem de kanserde ilaç direnci mekanizmalarının aydınlatılabilmesi açısından önem taşımaktadır.

Identification of the phenomenon of anastasis in breast cancer cells

Aim: This study aimed to define the expression changes and potential roles of the apoptosis-related genes in the anastasis process in breast cancer, which is one of the most common cancer types. Materials and Methods: Different types of breast cancer cell lines (MCF7 and MDA-MB-231), breast cancer stem cells, and healthy breast cell line (MCF10A) were used. Apoptotic and anastatic cell percentages were determined by the Annexin V test and flow cytometry. Gene expression changes in anastatic cells compared to apoptotic cells were determined by the qRT-PCR and 2-ΔΔCt method. The pathways and biological processes of genes that show significant changes were determined using the STRING v11.5 database. Results: In all cell groups, it was determined that the percentage of apoptosis increased as a result of ethanol application, and the percentage of apoptotic cells decreased with the removal of the apoptosis-inducing factor. The change in the percentage of apoptotic cells between the control, apoptosis, and anastasis groups was determined the most in MCF7 cells. Consistently, expression changes were determined in the largest number of genes in this cell line. CASP7 and APAF1 genes downregulated in all cell lines. In all cell groups, it was determined that anastasis affects Cysteine-type endopeptidase activity involved in execution phase of apoptosis (GO_ID: 0043027), and drug resistance-related pathways (KEGG_ID: hsa01524). Conclusion: The definition of the interaction of the anastasis phenomenon in cells with apoptosis regulatory mechanisms is important in terms of elucidating both the oncogenic transformation of healthy cells and the mechanisms of drug resistance in cancer.

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