骨髓增生异常综合征造血细胞生物学指标检测的意义

The implications of the biological indices examination of bone marrow cells in myelodysplastic syndromes

目的 研究骨髓增生异常综合征 (MDS)造血细胞的生物学特性改变及与疾病进展的关系。方法 对 37例MDS患者骨髓细胞进行染色体核型分析、CFU GM体外半固体琼脂培养、增殖细胞核抗原 (PCNA)表达、细胞周期分析以及造血克隆性检测 ,观察其与MDS疾病进展的关系。结果 ①MDS染色体异常检出率 31% ,随访过程中染色体核型正常的 9例中有 2例、异常的 5例中有 3例转化为急性髓系白血病 (AML)。②MDS患者骨髓细胞体外CFU GM集落产率减少 ,集簇产率增加。43%的患者CFU GM产率极低或无生长。随访中 9例CFU GM集簇生长为主的患者中 6例疾病进展或转化为AML ;6例集落、集簇不生长和 3例生长基本正常者中仅 1例转化为急性白血病。③MDS患者骨髓细胞PCNA表达明显升高 ,RAEB/RAEB t的PCNA表达较RA/RAS高。④MDS患者骨髓细胞溴脱氧尿嘧啶 (BrdU)标记指数减低 (5 .0 7% ) ,DNA合成期时间和潜在倍增时间延长 ,分别为 8.5 7h和 8.6 9d ,并与病程进展相关。⑤ 7例女性HUMARA等位基因为杂合子的RA患者中 ,2例为多克隆造血 ,5例为单克隆造血。 3例RAEB和 1例MDS/AML均为单克隆造血。结论 随着MDS病程进展 ,骨髓中原始细胞增多 ,造血细胞生物学特性呈现以下的演变趋势 :造血转变为单克隆性 ;体外培养CFU

Objective To explore the biological characteristics of the hematopoietic cells in myelodysplastic syndromes(MDS) and the relationship between the biological indices and the disease progressing. Methods R banding technique for karyotype analysis, in vitro semi solid culture for CFU GM growth patterns, flow cytometry after monoclonal antibody labeling for proliferation cell nuclear antigen (PCNA) expression, flow cytometry after in vitro bromodeoxyuridine(BrdU) incorporation for cell cycle kinetics analysis, and X linked HUMARA gene polymorphism assay for hematopoiesis clonality analysis were carried out in 37 MDS patients. Results ①Cytogenetic abnormalities were detected in 31% of the MDS patients and AML transformation occurred in 2 of 9 with normal and 3 of 5 with abnormal chromosomal karyotypes. ②The mean CFU GM colony yield was decreased and cluster yield increased in the MDS patients. Disease progression in 6 of 9 with more CFU GM cluster yields, while only 1 of 9 with either normal or no CFU GM yields transformed into AML. ③Percentages of PCNA positive BMMNCs were significantly higher in MDS patients than in normal control (44.22% vs 26.82%, P<0.01). RAEB/RAEB t subtypes had significantly higher PCNA positive cells (P<0.01). ④The mean BrdU labeling index of MDS marrow cells was lower (5.07%), the mean DNA synthesis time (Ts) and potential doubling time(Tpot) of MDS patients were longer than that of normal control (P<0.01), and the increased Ts and T pot were related to the disease progre ssion. ⑤Monoclonal pattern of BMMNCs was detected in 5 of 7 heterozygous female RA patients and the other 2 were polyclonal hematopoiesis. All 3 RAEB and 1 MDS/AML were monoclonal hematopoiesis. Conclusions With disease progressing and bone marrow blasts increasing, the biological nature of the hematopoietic cells in MDS exhibited a tendency to transformation as follows: hematopoiesis converting into clonal,in vitro CFU GM growth pattern approaching leukemic, PCNA expression increasing, and cell cycle prolonging.