IR spektroskopi kullanılarak in vitro meme kanser kök hücrelerinin araştırılması

Günnur Güler; Ümmü Güven; Eda Açıkgöz; Gülperi Öktem

doi:10.19161/etd.790394

EN TR

Investigation of breast cancer stem cells in vitro by using IR spectroscopy

Abstract

Aim: Cancer stem cells (CSCs) lead to tumor initiation, progression, relapse, metastasis and therapeutic resistance due to the ability of tumor to self-renewal and differentiate into other cell types. Therefore, the characteristic features of breast CSCs need to be determined in detail. The aim of this study was to investigate the differences in cell biochemistry at the molecular level by using Fourier transform infrared (FTIR) spectroscopy after breast CSCs were isolated with flow cytometry. Materials and Methods: Breast CSCs with CD44+/CD24- surface marker properties in the MCF-7 cancer cell lines were isolated by using flow cytometry (FACS). MCF10A, MCF-7 breast cancer cell line (cancer non-stem cells or non-CSCs) and breast CSCs were re-suspended into 0.9% NaCl, and each cell type was measured with the FTIR spectrometer. Results: The portion of breast CSCs with CD44+/CD24- surface marker properties in MCF-7 was 2.0-2.3%. In the FTIR spectra, spectral similarities and differences among breast CSCs, non-CSCs and healthy cells were determined. In breast CSCs, the lipid and protein signals are quite strong accompanied with an increased cell membrane fluidity and dynamics. When non-CSCs are compared with healthy cells, a less amount of both α-helical proteins and DNA is detected while an increase in the signals of negatively charged carboxyl groups is noticed. These data clearly show that breast CSCs exhibit a very different profile in terms of structure, content and dynamics of cellular macromolecules compared to both non-CSCs and healthy cells. Conclusion: Drug studies (targeted therapy, drug-action mechanism) can be performed by examining small changes in the molecular structure and content of breast CSCs. This study shows that FTIR spectroscopy can be used in advanced cell studies as well as in the analysis of biological samples in medical field due to rapid, label-free and accurate measurement without complex sample preparation procedures.

Keywords

IR spektroskopi kullanılarak in vitro meme kanser kök hücrelerinin araştırılması

Öz

Amaç: Kanser kök hücreleri (KKH), tümör içinde kendi kendilerini yenileme ve diğer hücre tiplerine farklılaşabilme kapasitesi sebebiyle tümörün başlaması, ilerlemesi, nüksetmesi, metastaz ve terapötik dirence yol açmaktadır. Bu nedenle, meme kanser kök hücrelerinin (MKKH) karakteristik özelliklerinin belirlenmesi gerekmektedir. Bu çalışmanın amacı, MKKH‟lerin akış sitometrisi ile izole edildikten sonra Fourier dönüşümlü kızılötesi (FTIR) spektroskopisi kullanarak hücre biyokimyasındaki farklılaşmalarının moleküler seviyede araştırılmasıdır. Gereç ve Yöntem: MCF-7 meme kanser hücre hattındaki CD44+/CD24- yüzey belirteç özelliği gösteren MKKH‟ler akış sitometrisi ile izole edilmiştir. MCF10A, MCF-7 kanser hücre (KH) hattı ve bu hattan izole edilen CD44+/CD24- yüzey belirteç özelliklerine sahip MKKH‟'ler %0,9 NaCI içerisine resuspanse edildikten sonra FTIR spektrometre ile ölçülmüştür. Bulgular: MCF-7 içerisindeki CD44+/CD24- yüzey belirteç özelliğine sahip KKH‟lerinin sort oranı %2,0-2,3 olarak belirlenmiştir. Elde edilen FTIR spektrumlarında, MKKH, meme kanser hücreleri (KH, non-KKH, bulk populasyon) ve sağlıklı hücreler arasında spektral benzerlikler ve farklılıklar tespit edilmiştir. MKKH‟lerde lipit ve protein sinyalleri daha güçlü olup hücre zarı akışkanlığı ve dinamiği fazladır. Sağlıklı hücreler ile kıyaslandığında, KH‟lerde α-helikal proteinler ve DNA sinyallerinde azalmaya karşın negatif yüklü karboksil gruplarından kaynaklanan sinyallerde artış gözlenmektedir. Bu veriler, MKKH‟lerin, sağlıklı ve KH‟lere kıyasla yapı, içerik ve dinamiği bakımından oldukça farklı bir profil sergilediğini göstermektedir. Sonuç: Bu çalışma, MKKH‟lerinin moleküler yapısı ve içeriğindeki değişikliklerin incelemesi vasıtasıyla terapötik hedefli ilaç çalışmaları yapılabileceğini ortaya koymaktadır. FTIR spektroskopisi boyar madde gerektirmeden, hassas ve hızlı ölçüm alınması, örnek hazırlamada kolaylık ve az miktarda örnek gerektirmesi sebebiyle ileri hücre çalışmalarında ve medikal alanda biyolojik örneklerin analizlerinde kullanılabileceği de gösterilmiştir.

Anahtar Kelimeler

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Details

Primary Language

Turkish

Subjects

Health Care Administration

Journal Section

Research Article

Authors

Günnur Güler ^*
0000-0002-8485-7372
Türkiye

Ümmü Güven
0000-0002-5427-263X
Türkiye

Eda Açıkgöz
0000-0002-6772-3081
Türkiye

Gülperi Öktem
0000-0001-6227-9519
Türkiye

Publication Date

September 30, 2020

Submission Date

September 4, 2019

Acceptance Date

November 2, 2019

Published in Issue

Year 2020 Volume: 59 Number: 3

DOI

https://doi.org/10.19161/etd.790394

IZ

https://izlik.org/JA42YG35HY

Cite

RIS / Bibtex

Vancouver

1.Günnur Güler, Ümmü Güven, Eda Açıkgöz, Gülperi Öktem. IR spektroskopi kullanılarak in vitro meme kanser kök hücrelerinin araştırılması. EJM. 2020 Sep. 1;59(3):149-54. doi:10.19161/etd.790394