[醫(yī)藥]骨髓增生異常綜合征論文：MDS、AA和AL患者骨髓細(xì)胞周期及增殖特征的研究

1、 骨髓增生異常綜合征論文：MDS、AA和AL患者骨髓細(xì)胞周期及增殖特征的研究【中文摘要】檢測(cè)骨髓增生異常綜合征(myelodysplastic syndrome, MDS)、再生障礙性貧血(aplastic anemia, AA)和急性白血病(acute leukemia, AL)患者骨髓單個(gè)核細(xì)胞(bone marrow mononuclear cells, BMMNC)的細(xì)胞周期分布及CD34、Ki67抗原的表達(dá)。方法選取2009年6月至2010年6月在山東大學(xué)附屬省立醫(yī)院就診的住院及門診患者68例,MDS組30例,其中難治性貧血(refractory anemia, RA)組18例,難治

2、性貧血伴原始細(xì)胞增多(refractory anemia with excess of blasts, RAEB)組12例；AA組22例,其中急性重型再障組7例,慢性非重型再障組15例；AL組16例,其中急性髓系白血病(acute myeloid leukemia, AML)組12例,急性淋巴細(xì)胞白血病(acute lymphoblastic leukemia, ALL)組4例。另外選取正常對(duì)照組18例(均為非血液病患者)。采用常規(guī)骨髓穿刺術(shù),抽取骨髓各2ml；應(yīng)用流式細(xì)胞儀(flow cytometry, FCM),通過(guò)碘化丙啶(propidium iodide, PI)細(xì)胞核染色及免疫熒光

3、雙標(biāo)法,對(duì)其BMMNC的細(xì)胞周期分布及CD34、Ki67的表達(dá)進(jìn)行檢測(cè)。結(jié)果1.MDS和AL患者與正常對(duì)照組患者骨髓單個(gè)核細(xì)胞中細(xì)胞周期分布及增殖特征表達(dá)的比較與正常對(duì)照組相比,MDS組和AL組患者其BMMNC中細(xì)胞周期G0/G1期、造血干/祖細(xì)胞(haemopoietic stem cell, HSC)CD34細(xì)胞、Ki67+細(xì)胞、CD34+Ki67+細(xì)胞、CD34+細(xì)胞中Ki67+細(xì)胞和Ki67+細(xì)胞中CD34+細(xì)胞各自的比例均顯著升高(P0.05),而CD34+細(xì)胞中Ki67+細(xì)胞比例顯著降低(P<0.05),且CD34+細(xì)胞、CD34+Ki67+細(xì)胞和Ki67+細(xì)胞中CD34+細(xì)

4、胞比例均顯著降低(P<0.01)。4.AA患者和RA患者骨髓單個(gè)核細(xì)胞中細(xì)胞周期分布及增殖特征表達(dá)的比較與AA組相比,RA組BMMNC中G0/G1期、CD34+細(xì)胞、Ki67+細(xì)胞、CD34+Ki67+細(xì)胞、CD34+細(xì)胞中Ki67+細(xì)胞和Ki67+細(xì)胞中CD34+細(xì)胞比例均顯著升高(P<0.01),而RA組的S和S+G2/M期細(xì)胞比例均顯著降低(P<0.01)。結(jié)論1.MDS患者骨髓細(xì)胞G1期阻滯且伴有造血干/祖細(xì)胞的高增殖特性,可解釋MDS患者骨髓細(xì)胞分化成熟障礙,在細(xì)胞形態(tài)學(xué)上可表現(xiàn)為病態(tài)造血,在臨床上可表現(xiàn)為骨髓增生活躍而外周血細(xì)胞減少,支持MDS的本質(zhì)是造血干/祖細(xì)

5、胞的惡性克隆性疾病。2.MDS與AL患者骨髓細(xì)胞存在G1期阻滯的同時(shí),都伴有造血干/祖細(xì)胞的高增殖特性。由RA進(jìn)展到RAEB,再由RAEB進(jìn)展到AL,BMMNC中CD34+細(xì)胞、CD34+細(xì)胞中Ki67+細(xì)胞比例均升高,G0/G1期細(xì)胞比例、Ki67+細(xì)胞比例亦升高,提示來(lái)源于造血干/祖細(xì)胞的惡性克隆性擴(kuò)增逐漸獲得優(yōu)勢(shì)而積聚,和/或其細(xì)胞本身質(zhì)的缺陷越來(lái)越嚴(yán)重；MDS的不同發(fā)展階段及其最終轉(zhuǎn)化為急性白血病,從細(xì)胞周期理論及其增殖特征分析,它們之間并無(wú)質(zhì)的差異,可能是同一疾病的不同發(fā)展階段,支持MDS和AL具有相似的發(fā)病機(jī)制。3.與正常對(duì)照組相比,AA患者骨髓細(xì)胞的細(xì)胞周期分布差異無(wú)統(tǒng)計(jì)學(xué)意義,

6、不支持AA是造血干/祖細(xì)胞惡性克隆性疾病,同時(shí)AA患者的造血干/祖細(xì)胞不僅數(shù)量減少,而且增殖活性減低,不同于MDS和AL患者。4.RA和AA患者相比,其骨髓細(xì)胞周期分布及增殖特性的表達(dá)差異均具有顯著性意義,可用于二者的診斷及鑒別診斷,尤其是BMMNC中CD34+Ki67+細(xì)胞比例兩者差異更大,用作本研究RA與AA的鑒別診斷指標(biāo)準(zhǔn)確率達(dá)100%?！居⑽恼縏o explore the cell cycle distributions and the expressions of CD34 and Ki67, the cell proliferation-related protein in b

7、one marrow mononuclear cells among myelodysplastic syndromes, aplastic anemia, and acute leukemia patients.METHODS Bone marrow aspirates were collected from 68 patients between June 2009 and June 2010 from the Department of Hematology, Provincial Hospital affiliated to Shandong University. The 68 ca

8、ses consist of 30 MDS, including 18 refractory anemia and 12 refractory anemia with excess blasts,22 AA, including 7 severe aplastic anemia and 15 none severe aplastic anemia, and 18 AL, including 12 acute myeloid leukemia and 4 acute lymphoblastic leukemia.18 healthy individuals (non-hematologic pa

9、tients) were used as normal control. Take 2 ml bone marrow from the patients. Propidium iodide and immunofluorescent double staining through flow cytometry were utilized to explore cell cycle distributions and the expression of CD34 and Ki67, the cell proliferation-related protein in bone marrow mon

10、onuclear cells.RESULTS1. Compared with the control group, the percentages of G0/G1 phase of cell cycle, haemopoietic stem/progenitor cells CD34+ cells, Ki67+ cells, CD34+Ki67+ cells, Ki67+ cells in CD34+ cells, and CD34+ cells in Ki67+ cells of BMMNC were all significantly increased in the MDS and A

11、L group (P0.05). The percentages of Ki67+ cells in CD34+ cells was significantly lower (P<0.05), as well as the percentages of CD34+ cells, CD34+Ki67+ cells, and CD34+ cells in Ki67+ cells of BMMNC were all significantly lower in the AA group than those in the control group (P<0.01).4. The per

12、centages of G0/G1 phase, CD34+, Ki67+, CD34+Ki67+, Ki67+ cells in CD34+, and CD34+ cells in Ki67+ cells were all significantly increased (P<0.01), although the proportions of S and S+G2/M phases were both significantly decreased in the RA group than the AA group (P<0.01).CONCLUSIONS1. Most of

13、the cells are kept at the G1 phase as well as the high proliferation of the haemopoietic stem/progenitor cells in MDS patients that can explain the dysdifferentiation and dysmaturity, which is the impaired peripheral blood cell production (cytopenias) as well as most commonly a hypercellular, dyspla

14、stic-appearing bone marrow. This greatly proves that the disorder of MDS might occur when the malignant damage of the haemopoietic stem/progenitor cells occurs.2. There are higher percentages of G1 phase as well as the high proliferations of the CD34+ cells in both MDS and AL patients. In our study,

15、 the ratios of G0/G1 phase, CD34+, Ki67+, and Ki67+ cells in CD34+cells were all significantly higher as RAEB progressed to AL as well as RA progressed to RAEB. This indicates that the disorder of MDS evolves the different stages and may eventually have the transformation to acute leukemia because o

16、f the expansion of malignant clonal cells from the haemopoietic stem/progenitor cells or because of more and more seriously qualitative defections in cells. Both of the two disorders may have the similar pathogenesis and may be the different stage of a common disease according to the theory of cell

17、cycle and cell proliferation.3. There was no significant difference on the cell cycle between patients with AA and normal controls. It proves that AA is an illness without the malignant clonal mutation of haemopoietic stem/progenitor cells. We also found that besides the haemopoietic stem/progenitor

18、 cells were lower, the expressions of proliferations of BMMNC were all significantly lower in the AA group. It is different from patients with MDS and AL.4. The disturbance of cell cycle and the expression of cell proliferation were all different in patients with RA from AA, which can be used for the diagnosis and differential diagnosis of them. It is most different on the ratio of CD34+Ki67+ cells that may be 100% accuracy for the diagnosis and differential diagnosis of RA and AA.【關(guān)鍵詞】骨髓增生異常綜合征 貧血,再生障礙性 急性白血病 細(xì)胞周期 細(xì)胞增殖【英文關(guān)鍵詞】Myel