ABSTRACT
Objective To explore the characteristics of morphology and immunology in acceleration phase and blastic phase of chronic myelogenous leukemia(CML).Methods Seventy-three cases of CML-BP bone marrow specimens were respectively conducted the morphology and related cell chemical dyeing observation for determining the FAB type.Flow cytometry was used to detect series immunological related antigens.Results Among 73 cases of FAB typing,there were 44 cases of CML-AML,21 cases of ALL and 8 cases.21 cases CML-ALL patients In the immunophenotyping by flow cytometry,among 21 cases of CML-ALL,there were 19 cases of B-ALL,2 cases of T-ALL,moreover 12 cases contained myeloid marker.Among 8 cases CML-HAL,the immunophenotypes were 6 cases of B+-My and 2 cases of T+ My.Among 44 cases CML-AML,15 cases contained T cell marker,and 2cases contained B cell marker,other cases had no cross-lineage expression.Among 73 cases of CML-BP,29 cases conducted the flow cytometry detection in the acceleration phase,in which 16 cases urgently changed to AML,and 13 cases to non-AML(9 cases of ALL and 4 cases of HAL).Among non-AML cases,2 cases had the simultaneous existence of myeloid primitive cells and precursor lymphocyte in the acceleration phase and other 9 cases were myeloid primitive cell or accompanied by lymphocyte marker.Conclusion Flow cytometry has a certain implication role for the direction and differentiation diagnosis of CML-BP.
ABSTRACT
Objective To investigate expression and regulatory mechanism of phosphatase and tensin hemology deleted on chromosome ten gene (PTEN) and cyclooxygenase-2 (COX-2) in human myeloid leukemia cells.Methods Thirty patients was collected from the First Hospital of Baoding and Second Affiliated Hospital of Hebei Medical University,including 10 chroni myeloid leukemiac (CML)patients in chronic phase (CML-CP),10 CML patients in blast crises (CML-BC) and 10 normal controls.The recombinated adenovirus containing green fluorescent protein (GFP) and PTEN ( Ad-PTEN-GFP) or empty vector (Ad-GFP) was transfected into human CML K562 cells.The growth of K562 cells and cell adhesion ability was observed by 3-[4,5-dimethyl-2-thiazolyl]-2,5-dip (MTF) assay; PTEN and COX-2 messenger ribonucleic acid (mRNA) levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR (FQ-PCR).PTEN and p-Akt protein levels were detected by western blot,and COX-2 protein were measured with cytochemical staining.ResultsThe mRNA expression levels of PTEN in CML-BC patients (0.022 ±0.021 ) were lower than CML-CP patients (1.134 ± 1.124) and normal control ( 1.059 ±0.595).The mRNA expression levels of COX-2 in CML-BC patients (0.761 ±0.418) were higher than CML-CP (0.211 ± 0.158) and normal control (0.165 ± 0.152).The growth of K562 cells was suppressed markedly,and the maximum growth inhibition rate was 38.67％ ± 4.30％ after transfected with PTEN gene,which was higher than Ad-GFP group (5.34％ ±0.31％,t =13.39,P ＜0.01 ).COX-2 mRNA expression levels in Ad-PTEN-GFP(0.013 ± 0.001 )were significantly lower than Ad-GFP group (0.199 ±0.018) and untransfected group (0.217 ± 0.021,F =499.45,P ＜ 0.01 ),and p-Akt as well as COX-2protein were also down-regulated after K562 cells transfected ( MOI =200) with wild type PTEN in three days.ConclusionPTEN may inhibit proliferation and adhesion ability of leukemia cell in myeloid leukemia via down-regulating COX-2 expression.
