Akciğer kanseri hastalarında dolaşımdaki tümör hücrelerini nasıl tespit ederiz? akım sitometrisi ile yapılan kısa bir çalışma

Year 2018, Volume: 3 Issue: 4, 179 - 189, 29.12.2018

Özen Özensoy Güler , Tuğba Uysal Ender Şimşek , Ahmet Çarhan

https://doi.org/10.26453/otjhs.410582

Abstract

Dolaşımdaki tümör hücreleri (CTC’ler)
karsinomların metastatik yayılımında önemli role sahiptir. Bu nedenle, son
yıllarda birçok kanser türünde hastalığın seyri ve tedavi etkinliğinin
anlaşılmasında yardımcı olabileceği düşünülen CTC’ler üzerine yoğunlaşılmıştır.
Teknik yaklaşımlardaki gelişmeler ile özellikle akciğer kanseri gibi dokularına
ulaşılması zor olan tümörlerde sıvı biyopsi olarak tüm kandan CTC tespitinin
değeri her geçen gün artmaktadır. Tanı anında sıklıkla metastaz yaptıkları
teşhis edilen akciğer kanseri, hem kadınlarda hem de erkeklerde kanser ölüm
nedenlerinin birincil sebebidir. Çalışmamızın amacı akım sitometrisi ile
akciğer kanserli hastaların periferik kan örneklerinde (7.5 mL) CTC tespitidir.
Zenginleştirme ve saptama adımlarından oluşan modifiye ettiğimiz yöntemimiz ile
CTC sayısının tespit edilmesi için 9 akciğer kanserli birey ve 9 sağlıklı birey
çalışmaya dahil edilmiştir. Uyguladığımız metotta zenginleştirme basamağı için
fikol yoğunluk gradiyent ayrımı ve immünomanyetik ayırma tekniği (CD45 negatif
seçilim) gerçekleştirilmiştir. Sonrasında, zenginleştirilmiş hücreler arasında
CTC’leri tespit etmek için, anti-epitelyal hücre adhezyon molekülünün ve
sitokeratinlerin ekspresyonuna dayanan çok parametreli akım sitometrisi ile
analiz yapılmıştır. Çalışmamız sonucunda akciğer kanserli hastaların tümünde
CTC gözlenirken, sağlıklı bireylerde gözlenmemiştir (Z=3.823; p<0.001).
Böylece, modifiye ettiğimiz metodun akciğer kanserinde CTC tespitinde
kullanılabilirliği gösterilmiştir.

Keywords

Dolaşımdaki tümör hücreleri, akım sitometresi, akciğer kanseri

How could we detect circulating tumor cells in lung cancer patients? A brief study by flow cytometry

Abstract

Circulating tumor cells (CTCs) play a
crucial role in the metastatic spread of carcinoma. Therefore, CTC has been
interest of a subject in the past few decades in terms of prognosis and
response to the therapy in several cancer diseases. Recent improvements in
technical approaches maintain to identify CTCs from whole blood have
demonstrated the potential value of CTC detection as a liquid biopsy especially
in those tumors where tissue accessibility is often challenging as in lung
cancer. Lung cancer is the most common cause of death from cancer worldwide in
both men and women which is commonly metastasize before it is diagnosed. The
aim of this study is to enumerate of CTCs in peripheral blood sample (7.5 mL)
of lung cancer patients by flow cytometry.  Our modified method which consists of
enrichment and detection steps get involved in 9 patients with lung cancer and
9 healthy volunteers. We performed a density-based ficoll gradient
centrifugation and a immunomagnetic separation technique (CD45 negative selection)
for the enrichment step.  Next,
multi-parameter flow cytometry based on the expression of anti-epithelial cell
adhesion molecule and cytokeratins was used to detect circulating tumor cells
among enriched cells. According to our results, 
circulating tumor cells were not detected on healthy volunteers but
circulating tumor cells were found in all of patients with lung cancer
(Z=3.823; p<0.001). We demonstrate that circulating tumor cells were
detectable in peripheral blood sample of lung cancer patients by our modified
method.

Keywords

Circulating tumor cells, flow cytometry, lung cancer

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