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

Öz

Objective: Determination the expression of P-glycoprotein is especially problematic for normal tissues because immunological 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 patients diagnosed with hematologic malignancy.
Methods: Our study included fifty patients diagnosed with acute myeloblastic leukemia and acute lymphoblastic leukemia, and twenty healthy controls who were admitted to Erciyes University Hematology-Oncology Hospital. The suspended cells from bone marrow samples of patients and the peripheral 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 lymphoblastic leukemia patients P-glycoprotein, in 5 of them CD34 expression were determined. A significant relation between P-glycoprotein and CD34 expressions in acute myeloblastic 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 measuring P-glycoprotein expression and suggests the possibility 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.
Key words: MDR-1 gene, AML, ALL, P-glycoprotein, flow cytometry

Anahtar Kelimeler

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Hematolojik Malignansilerde P-Glikoprotein Ekspresyonunun Akım Sitometri İle Belirlenmesi

Öz

Amaç: Duyarlılık açısından sınırlı olan immünolojik yöntemler kullanılarak özellikle normal dokulardaki P-glikoprotein ekpresyonunun belirlenmesi sorun oluşturmaktadır. Bu çalışmada hematolojik malignansi tanısı almış ve tedaviye yanıt vermeyen hastalarda P-glikoprotein ve CD34 ekpresyonunun değerlendirilmesi için, P-glikoprotein and CD34 ekpresyonunun akım sitometri ile belirlenmesini amaçladık.Yöntemler: Erciyes Üniversitesi Mehmet-Kemal Dedeman Hematoloji-Onkoloji Hastanesine başvuran akut miyeloblastik 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 örneklerinden elde adilen süspanse hücreler, P-glikoprotein fikoeritrin ile ve CD34 FITC ya da PerCP Cy 5.5 antikorları ile işaretlendi ve daha sonra yüzey ekspresyonları akım sitometri ile ölçüldü.Bulgular: Çalışmamızda 30 akut miyeloblastik lösemi hastasının 6’sında (%20) P-glikoprotein ve CD34 ekspresyonu tespit edilirken, 20 akut lenfoblastik lösemi hastasının 6’sında (%30) P-glikoprotein, 5’inde (%25) CD34 ekspresyonu tespit edildi. Akut miyeloblastik lösemi ve akut lenfoblastik lösemi kemik iliği örneklerinde P-glikoprotein ve CD34 ekspresyonları arasında anlamlı bir ilişki olduğu görüldü. Sonuç: Bulgular, akım sitometri yöntemi ile P-glikoprotein ekspresyonunun akut miyeloblastik lösemi ve akut lenfoblastik lösemi kemik iliği örneklerinde moleküler yöntemlerden daha hızlı, güvenilir ve doğru bir şekilde ölçülebildiğini, P-glikoprotein ile CD34 ekspresyonları arasında anlamlı bir ilişki olabileceğini göstermiştir. Yüzeyinde CD34 eksprese eden blast hücrelerinin aynı zamanda P-glikoprotein eksprese etmeleri hücrelerdeki çoklu ilaç direnci geninin daha çok olgunlaşmamış hücrelerde aktif olduğunu göstermektedir

Anahtar Kelimeler

• Çoklu ilaç direnci-1 geni, AML, ALL, P-glikoprotein, akım sitometri

Kaynakça

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