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

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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