Detection Of BCR-ABL Fusion Gene With Endothelial Progenitor Cells In Chronic Myelogenous Leukemia And Its Clinical Significance

[Objectives] To detect the expression level of BCR/ABL fusion gene with endothelial progenitor cells (EPCs) in chronic myelogenous leukemia(CML); and to investigate the role of EPCs in disease progression.[Methods] EPCs isolated from bone marrow of CML patients for the study.Isolated EPCs with bone marrow were selected by density gradient centrifugation. The morphologic features of EPCs were observed by microscopy, the cell growth rate was calculated by cell counting, and the cells were identified by immunofluorescence staining. The expression of BCR-ABL fusion gene was examined by RQ-PCR and FISH; And the BCR-ABL fusion gene with EPCs correlation with clinical data were performed.[Results]1. The bone marrow-derived EPCs are adherent and clone-like growth after being cultured 3d, the number of cells increase gradually at 6d,80% cells fusion after 7d.The cells showed a typical cobblestone-like arrangement at 14d.The result of EPCs by immunofluorescence shows that they can stain Dil-acLDL and FITC-UEA-1.The growth curves of bone marrow-derived EPCs of CML patients are S-shaped.2. Two of the BCR-ABL fusion gene in endothelial progenitor cells (EPCs) isolated from bone marrow of CML patients were positive by RQ-PCR, NQ values significantly decreased after treatment.3. The BCR-ABL fusion gene in endothelial progenitor cells (EPCs) isolated from bone marrow of CML patients were negative by FISH.4. The positive rate of BCR-ABL (p210)fusion gene in the CML patients with bone marrow tumor cells,which was significantly different from that of in the EPCs isolated from bone marrow of the CML patients (P<0.05).5. The positive rate of BCR-ABL(p210) fusion gene in the EPCs isolated from bone marrow of complex chromosome patients, which was significantly different from that of ordinary ph chromosome patients(P<0.05).6. The positive rate of BCR-ABL(p210) fusion gene in EPCs isolated from bone marrow of high-risk group of patients had no significant difference than that of low-risk group(P>0.05).7. The patients of BCR-ABL(p210) (+) was significantly different from that of BCR-ABL(p210) (-) between the WBC(t=2.986, p=0.008)、LDH(t=2.920, p=0.009) (P<0.05).Which had no significant difference between the platelets (t=0.237, p=0.815)、HB (t=-0.821, p=0.422)、β2-MG (t=0.362, p=0.721) (P>0.05).[Conclusion] The EPCs can be isolated and cultured from the bone marrow of CML patients; The NQ values of BCR-ABL fusion gene in endothelial progenitor cells (EPCs) isolated from bone marrow of CML patients is lower than before treatment, it is concluded that the EPCs may participate in patients with CML,and the BCR-ABL fusion gene were detected in the EPCs of complex chromosome karyotype patients,which possibly correlate to the EPCs are associated with the prognosis of CML patients.