Çeşitli lösemi tiplerinde tedavi öncesi ve sonrasında kromozom anomalilerin GTG-bantlama tekniği ve ABL-BCR füzyon gen ekspresyonunun RT-PCR yöntemi ile tespiti

Abstract

ÖZET Bu çalışmada değişik lösemi türlerinde, sitogenetik ve moleküler genetik yöntemlerle, kromozom anomalileri incelenmiştir. 280 olgunun tedavi öncesinde kemik iliği veya periferik kan örneklerinden hücre kültürü yapılmıştır. Kültürleri üremeyen, tedavi esnasında ölen, tedaviyi terk eden veya remisyona girmeyen hastalar elendikten sonra kalan 70 olgu bu çalışmanın esas materyalini oluşturmaktadır. Bu 70 olgudan 30'u Akut miyeloid lösemi (AML), 9'u Akut lenfoid lösemi (ALL), 29'u Kronik miyeloid lösemi (KML) ve 2'si Miyelodisplastik sendrom (MDS) göstermekteydiler. Yetmiş hastanın sitogenetik analizi ile, tedavi öncesinde 34 hastada kromozom anomalisi bulunmuşken, tedavi sonrasında yeni çoğalan hücrelerde bu 34 kromozom anomalisinden 22 tanesi görülmemiş, 12 tanesinin ise varlığını devam ettirdiği saplanmıştır. Tedavi öncesinde sitogenetik yöntemle belirlenen kromozom anomalisi türlerinin lösemi gruplarına göre dağılımı şöyledir : Otuz AML olgusundan; ikisinde translokasyon t(8;21), birinde trizomi 4, birinde trizomi 1 1 bulunmuştur. Dokuz ALL olgusundan, birinde translokasyon t(9;22), ikisinde delesyon del(12)(p), birinde monozomi 7, birinde trizomi 22, birinde trizomi lq bulunmuştur. Yirmidokuz KML olgusundan, yirmiüçünde translokasyon t(9;22) anomalisi bulunmuştur, tki MDS olgudan; birinde monomozi 7 bulunmuştur. Moleküler genetik yöntem ile yalnız translokasyon t(9;22) anomalisi araştırılmış olup, tedavi öncesinde 70 hastanın 33 tanesinde t(9;22) anomalisi bulunmuşken, tedavi sonrasında yeni çoğalan hücrelerde 29 hastada bu anomalinin varlığını devam ettirdiği görülmüştür. Moleküler genetik yöntemle tedavi öncesinde translokasyon t(9;22) anomalisi belirlenen hastaların değişik lösemi gruplarına göre dağılımı şöyledir ; KML grubunda yirmidokuz hastada, AML grubunda bir hastada, ALL grubunda üç hastada bulunmuştur. Translokasyon t(9;22) anomalisinin belirlenmesi açısından; sitogenetik yöntem ile moleküler genetik analiz yönteminin karşılaştırılması sonucunda, moleküler genetik yöntemin bu anomaliyi saptamak açısından daha etkili olduğu kanısına varılmıştır. Anahtar kelimeler ; lösemi, sitogenetik, RT-PCR, kromozom anomalisi

SUMMARY INVESTIGATING EXPRESSION OF ABL-BCR FUSION GENE BY RT-PCR AND CHROMOSOMAL ANOMALIES BY GTG-BANDING TECHNIQUE IN DIFFERENT TYPES OF LEUKEMIA BEFORE AND AFTER TREATMENT In this study, the chromosome abnormalities in different types of leukemia were investigated with cytogenetic and molecular genetic methods. Of 280 patients' bone marrow or peripheral blood cell cultures were prepared before the treatment. The patients who wasn't grow lymphocyte cultures or not in remission, and leave the treatment and also die during the treatment were eliminated. After the elemination, 70 patients were the main material of this study. The thirty of 70 patients had acute myeloid leukemai (AML), 9 of them had acute lymphoid leukemia (ALL), 29 of them had chronic myeloid leukemia (CML) and 2 of them had myelodysplastic syndrome (MDS). Before and after the treatment, these 70 cases were evaluated with cytogenetic method (GTG-banding) and molecular genetic method (RT-PCR) if they have chromosome abnormality or not. With the cytogenetic method before treatment, 34 chromosomal anomalies were found in 70 patients. After treatment only 12 of 34 anomalies had continued. The distribution of chromosome anomalies which were evaluated before treatment with cytogenetic methodare as follows ; among thirty AML cases, two patient had the translocation t(8;21), one patient had trisomy 4, one patient had trisomy 1 1. Among nine ALL cases, one patient had the translocation t(9;22), two patients had deletion del(12p), one patient had monosomy 7, one patient had trisomy 22, and one patient had trisomy lq. Among twenty-nine CML cases, 23 patients had the translocation t(9;22). Among two MDS cases, one patient had monosomy 7. The anomaly of translocation t(9;22) which was investigated with the molecular genetic method had been found in 33 of 70 patients at the beginning of treatment. This anomaly was stable 29 of 33 after treatment. The distribution of t(9;22) anomaly which was evaluated before treatment with molecular method is as follows ; among thirty AML cases, one patient, among nine acute lymphoid leukemia cases, three patients, among twenty-nine CML cases, twenty-nine patients had the translocation. Before the treatment the anomaly of translocation t(9;22) found with the molecular genetic method are according to leukemia groups following : from the thirty AML cases in KVRUWone patient, from nine acute lymphoid leukemia cases in three patients, from twenty-nine CML cases in twenty-nine patients. In this study it has been found that the molecular genetic method is much more sensitive than cytogenetic method for detecting the anomaly of translocation t(9;22). Key words ; leukemia, cytogenetic, RT-PCR, III