Influence of PD-0332991 treatment on cell cycle regulatory genes in breast cancer stem cells

Hasan Onur Çağlar; Sunde Yılmaz Süslüer; Cumhur Gündüz; Ayfer Haydaroğlu

doi:10.19161/etd.414615

TR EN

Meme kanseri kanser kök hücrelerinde PD-0332991 uygulanmasının hücre döngüsü düzenleyici genler üzerine etkisi

Öz

Amaç: Palbociclib (PD-0332991) siklin bağımlı kinaz 4/6 kompleksi için bir inhibitördür ve luminal ER+ meme kanserlerinde daha fazla aktivite sergilemektedir. Ancak, meme kanseri kök hücrelerinin (MKKH) PD-0332991 uygulamasına olan hassasiyeti ve hücre döngüsü düzenleyici genleri üzerine PD-0332991’in etkisi MKKH’leri için açıklığa kavuşmamıştır. Bu çalışma, MKKH’lerinin PD-0332991 uygulamasına olan yanıtın belirlenmesini amaçlamaktadır.

Gereç ve Yöntem: Meme kanseri hücre hatları üzerinde deneysel bir in vitro çalışma tasarlanmıştır. Bu çalışmada, MCF-7 ve MKKH hücre hatları kullanıldı. Suda çözünür tetrazolyum tuzu-1 (WST-1) testi sitotoksisite deneyi için kullanıldı. Hücre döngüsü dağılımı ve apoptosis, 48. saat IC50 değerlerine göre flow sitometri ile incelendi. CDKN1A, CHEK1, CDKN2A, CDC25A ve CCND1 genlerinin ifade profillerinin belirlenmesinde Real-Time PCR kullanıldı.

Bulgular: Her iki hücre hattında, PD-0332991 hücre proliferasyonunu azalttı. Her iki hücre hattı için G0/G1 tutuklanması belirlendi. PD-0332991’in apoptotik etkisi MCF-7 ve MKKH hücrelerinde bulunmadı. MCF-7 hücrelerinde, CDKN1A, CDKN2A ve CCND1 ifade düzeyleri sırasıyla 3,11; 3,21 ve 1,05 kat arttı. CHEK1 ve CDC25A ifade düzeyleri sırasıyla 4,75 ve 3,73 kat azaldı. MKKH’lerinde CDKN1A, CHEK1, CDKN2A ve CCND1 ifade düzeleri sırasıyla 1,15; 2,01; 1,32 ve 1,68 kat azaldı. MKKH’lerinde, CDC25A geni ifadesi bulunmadı.

Sonuç: Bu çalışmada, MKKH ve meme kanseri hücreleri arasında PD-0332991’in hücre döngüsü düzenleyici genler için farklı ifade profillerine neden olduğu gözlenmiştir.

Anahtar Kelimeler

PD-0332991,meme tümörleri,neoplastik kök hücreler,G1 hücre döngüsü bloğu,hücre döngüsü.

Influence of PD-0332991 treatment on cell cycle regulatory genes in breast cancer stem cells

Abstract

Aim: Palbociclib (PD-0332991) is an inhibitor for cyclin-dependent kinase 4/6 complex and exhibits more activity in luminal ER+ breast cancer. However, sensitivity of breast cancer stem cells (BCSCs) to PD-0332991 treatment and expression patterns of cell cycle regulatory genes after PD-0332991 treatment in BCSCs are still unclear. This study aims to determine response of BCSCs to PD-0332991 treatment.

Materials and Methods: An experimental in vitro study was designed on breast cancer cell lines. MCF-7 and BCSCs cell lines were used in this study. Water soluble tetrazolium salt-1 (WST-1) test was used for the cytotoxicity assay. Cell cycle distribution pattern and apoptosis were examined with flow cytometry according to IC50 values at 48th h. Real-Time PCR was used to detect expression profiles of CDKN1A, CHEK1, CDKN2A, CDC25A, and CCND1 genes.

Results: PD-0332991 decreased cell proliferation in both cell lines. G0/G1 arrest was detected for both cell lines. There was no apoptotic effect of PD-0332991 on MCF-7 cells and BCSCs. In MCF-7 cells, expression levels of CDKN1A, CDKN2A, and CCND1 genes were increased as 3.11, 3.21, and 1.05 folds, respectively. Expression levels of CHEK1 and CDC25A genes were decreased as 4.75 and 3.73 folds, respectively. In BCSCs, expression levels of CDKN1A, CHEK1, CDKN2A, and CCND1 were decreased as 1.15, 2.01, 1.32, and 1.68 folds, respectively. No expression of CDC25A gene was found in BCSCs group.

Conclusion: In this study, it was observed that PD-0332991 leads to different expression profiles for cell cycle regulatory genes between BCSCs and breast cancer cells.

Keywords

PD-0332991,breast neoplasms,neoplastic stem cells,G1 cell cycle arrest,cell cycle

Kaynakça

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Ayrıntılar

Birincil Dil

İngilizce

Konular

Sağlık Kurumları Yönetimi

Bölüm

Araştırma Makalesi

Yazarlar

Hasan Onur Çağlar ^*
0000-0002-3637-4755
Türkiye

Sunde Yılmaz Süslüer
0000-0002-0535-150X
Türkiye

Cumhur Gündüz
0000-0002-6593-3237
Türkiye

Ayfer Haydaroğlu
0000-0001-5709-0981
Türkiye

Yayımlanma Tarihi

1 Mart 2018

Gönderilme Tarihi

22 Ağustos 2016

Kabul Tarihi

28 Şubat 2018

Yayımlandığı Sayı

Yıl 2018 Cilt: 57 Sayı: 1

DOI

https://doi.org/10.19161/etd.414615

IZ

https://izlik.org/JA77FH62RB

Kaynak Göster

RIS / Bibtex

Vancouver

1.Hasan Onur Çağlar, Sunde Yılmaz Süslüer, Cumhur Gündüz, Ayfer Haydaroğlu. Influence of PD-0332991 treatment on cell cycle regulatory genes in breast cancer stem cells. ETD. 01 Mart 2018;57(1):36-45. doi:10.19161/etd.414615