BERKAY SARAYMEN, AYSUN ÇETİN, Behzat ÇİMEN, İHSAN ÇETİN, MUSTAFA YAVUZ KÖKER, Bülent ESER

Determination of P-Glycoprotein Expression by Flow Cytometry in Hematological Malignancies

Amaç: Duyarlılık açısından sınırlı olan immünolojik yöntem- ler kullanılarak özellikle normal dokulardaki P-glikoprotein ekpresyonunun belirlenmesi sorun oluşturmaktadır. Bu çalış- mada hematolojik malignansi tanısı almış ve tedaviye yanıt vermeyen hastalarda P-glikoprotein ve CD34 ekpresyonu- nun değerlendirilmesi için, P-glikoprotein and CD34 ekpres- yonunun akım sitometri ile belirlenmesini amaçladık. Yöntemler: Erciyes Üniversitesi Mehmet-Kemal Dedeman Hematoloji-Onkoloji Hastanesine başvuran akut miyeloblas- tik lösemi ile akut lenfoblastik lösemi tanısı almış 50 hasta ve yirmi kontrol çalışmaya dâhil edildi. Hastaların kemik iliği örnekleri ile kontrol grubundan alınan periferik kan örnekle- rinden elde adilen süspanse hücreler, P-glikoprotein fikoerit- rin ile ve CD34 FITC ya da PerCP Cy 5.5 antikorları ile işa- retlendi ve daha sonra yüzey ekspresyonları akım sitometri ile ölçüldü. Bulgular: Çalışmamızda 30 akut miyeloblastik lösemi has- tasının 6sında (%20) P-glikoprotein ve CD34 ekspresyonu tespit edilirken, 20 akut lenfoblastik lösemi hastasının 6sında (%30) P-glikoprotein, 5inde (%25) CD34 ekspresyonu tespit edildi. Akut miyeloblastik lösemi ve akut lenfoblastik lösemi kemik iliği örneklerinde P-glikoprotein ve CD34 ekspresyon- ları arasında anlamlı bir ilişki olduğu görüldü. Sonuç: Bulgular, akım sitometri yöntemi ile P-glikoprotein ekspresyonunun akut miyeloblastik lösemi ve akut lenfob- lastik lösemi kemik iliği örneklerinde moleküler yöntemlerden daha hızlı, güvenilir ve doğru bir şekilde ölçülebildiğini, P-gli- koprotein ile CD34 ekspresyonları arasında anlamlı bir ilişki olabileceğini göstermiştir. Yüzeyinde CD34 eksprese eden blast hücrelerinin aynı zamanda P-glikoprotein eksprese et- meleri hücrelerdeki çoklu ilaç direnci geninin daha çok olgun- laşmamış hücrelerde aktif olduğunu göstermektedir.

Hematolojik Malignansilerde P-Glikoprotein Ekspresyonunun Akım Sitometri İle Belirlenmesi

Objective: Determination the expression of P-glycoprotein is especially problematic for normal tissues because immuno- logical methods are limited in terms of sensitivity. We aimed to determine the expression of P-glycoprotein and CD34 by flow cytometry, and to evaluate the level of expression of P- glycoprotein and CD34 with unresponsive to treatment in pa- tients diagnosed with hematologic malignancy. Methods: Our study included fifty patients diagnosed with acute myeloblastic leukemia and acute lymphoblastic leuke- mia, and twenty healthy controls who were admitted to Erci- yes University Hematology-Oncology Hospital. The suspend- ed cells from bone marrow samples of patients and the pe- ripheral blood samples of healthy people were marked with P-glycoprotein phycoerythrin and CD34 FITC or PerCP Cy 5.5; and then surface expression was measured by means of flow cytometry. Results: In 6 of 30 acute myeloblastic leukemia patients P- glycoprotein and CD34 expression, in 6 of 20 acute lympho- blastic leukemia patients P-glycoprotein, in 5 of them CD34 expression were determined. A significant relation between P-glycoprotein and CD34 expressions in acute myeloblas- tic leukemia and acute lymphoblastic leukemia bone marrow samples was reported. Conclusion: Our data indicate that flow cytometry is more reliable, precise and faster than molecular methods for mea- suring P-glycoprotein expression and suggests the pos- sibility of a significant relationship between P-glycoprotein and CD34 expressions in acute myeloblastic leukemia and acute lymphoblastic leukemia bone marrow samples. The blast cells expressing CD34 on their surface along with P- glycoprotein simultaneously show that multi drug resistance 1 gene is mostly active in immature cells.

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