恶性造血细胞表面Flt3受体的表达及其对Flt3配体的反应性研究

目的 :研究恶性造血细胞表面Flt3受体的表达 ,TNFα和地塞米松 (DXM )对Flt3受体表达的作用及重组人Flt3配体 (rhFL)对恶性造血细胞增殖的影响。方法 :用流式细胞仪测定 18株体外培养的恶性造血细胞株细胞表面Flt3受体 ,用MTT法测定rhFL对恶性造血细胞增殖的影响。结果 :5株细胞表面存在Flt3受体。B淋巴瘤细胞株Raji、Daudi及多发性骨髓瘤细胞株 82 6 6表达高水平Flt3受体 ;髓系白血病细胞株HL 6 0和多发性骨髓瘤细胞株XG 6表达低水平的Flt3受体。用 10 -6mol/LDXM培养 2 4h后 ,上述细胞Flt3受体表达降低 ;用2 0ng/mlTNFα培养 2 4h后 ,Raji和 82 6 6细胞Flt3受体表达降低 ,HL 6 0和XG 6细胞Flt3受体表达增高 ,Daudi细胞Flt3受体表达不受影响。rhFL在 10～ 10 0ng/ml浓度时 ,刺激Raji和HL 6 0细胞的增殖 (P <0 0 5 ) ,但对绝大多数的恶性造血细胞体外无刺激作用。结论 :多数恶性造血细胞不表达Flt3受体 ;rhFL也不引起此类细胞的增殖。地塞米松降低Flt3受体的表达 ,有可能用于防止或降低FL对部分恶性造血细胞的刺激作用 ,使FL的应用更为安全。     

Neuropilin-1某因在髓细胞白血病和正常人骨髓基质细胞中的表达(英文)

目的了解 neuropilin-1(NP-1)基因在髓细胞白血病(AML 和 CML)患者及正常人骨髓基质细胞中的表达情况。方法收集12例 AML、14例 CML 和20例正常对照骨髓标本,分离单个核细胞。进行体外长期培养,收集贴壁细胞(骨髓基质细胞)。利用逆转录-聚合酶链反应分别检测3组骨髓基质细胞中 NP-1基因的表达。结果成功建立了 AML、CML 和正常人骨髓基质细胞培养方法,NP-1基因在 AML、CML 骨髓基质细胞中的表达率分别为47.1%和50%,明显低于正常对照(85%)。结论 NP-1基因可表达于部分 AML、CML 和大部分正常人的骨髓基质细胞中,其在髓细胞白血病患者骨髓基质细胞中的低表达,可能与其调节造血功能异常有关。     

Neuropilin-1基因在髓细胞白血病和正常人骨髓基质细胞中的表达

目的了解neuropilin-1(NP-1)基因在髓细胞白血病(AML和CML)患者及正常人骨髓基质细胞中的表达情况。方法收集12例AML、14例CML和20例正常对照骨髓标本,分离单个核细胞。进行体外长期培养,收集贴壁细胞(骨髓基质细胞)。利用逆转录-聚合酶链反应分别检测3组骨髓基质细胞中NP-1基因的表达。结果成功建立了AML、CML和正常人骨髓基质细胞培养方法,NP-1基因在AML、CML骨髓基质细胞中的表达率分别为47.1%和50%,明显低于正常对照(85%)。结论NP-1基因可表达于部分AML、CML和大部分正常人的骨髓基质细胞中,其在髓细胞白血病患者骨髓基质细胞中的低表达, 可能与其调节造血功能异常有关。     

DIG掺入PCR半定量检测AML中Flt3基因的表达

目的：观察Ｆｌｔ３基因在ＡＭＬ（急性髓细胞性白血病）病人骨髓中的表达水平及正常对照骨髓１外周血表达水平的变化及其与病情、疗铲的关系。方法：应用白细胞Ｆｅｒｒｃｏｌｌ不连续密度梯度分离法分离出骨髓中恶性细胞,采用ＤＩＧ掺入半定量ＲＴ－ＰＣＲ,检测Ｆｌｔ３基因在２２例ＡＭＬ初治病人,５例ＡＭＬＣＲ病人的骨髓、１０例正常人和非恶性血液病病人１３例外周血、１７例骨髓以及５种白血细胞株中的表达水平。结果：５     

急性髓系白血病Evi1基因表达的研究

目的 :探讨急性髓系白血病中 Evi1基因表达及意义。方法 :应用 RT- PCR方法检测了 94例急性髓系白血病 (AML ,包括初治 4 9例 ,复发 2 3例 ,缓解 2 2例 )患者及 1 0名正常对照 Evi1基因的表达。结果 :1 0名正常人骨髓单个核细胞中无 Evi1 m RNA的表达。AML 患者 Evi1基因总的阳性表达率为 1 8.1 % (1 7/94 ) ,M5型未见表达 ,其中初治、复发、缓解组的 Evi1基因阳性率分别为2 6 .5 %、1 7.4 %、0 ,初治和复发组差别无显著性 (P=0 .5 5 4 )。M3患者中 Evi1、PML /RARα双阳性组与 PML /RARα单阳性组相比复发率高 (P<0 .0 5 ) ,早期死亡率高。 Evi1阳性的 AML患者多数生存期短。结论 :Evi1基因是一个原癌基因 ,在髓系白血病的发病中起重要作用 ,是髓系白血病预后不良的一个指标。     

急性髓细胞白血病患者外周血及骨髓CD87表达的改变

目的探讨急性髓细胞白血病（AML）患者外周血及骨髓CD87表达的改变。方法应用流式细胞术（FCM）检测了30例AML患者（M1 2例,M2 6例,M4 8例,M5 14例）外周血及骨髓CD87表达,应用配对t检验分析外周血及骨髓CD87表达的改变。结果14例M5患者骨髓CD87表达为9.47％～80.32％,外周血CD87表达为11.49％。87.46％;8例M4患者骨髓CD87表达为14．27％～46．28％,外周血CD。表达为14.79％～47.19％;6例M2患者骨髓CDg7表达为4．67％～34．26％,外周血CD＂表达为8.96％。39.78％;2例M,患者骨髓CDg7表达为3．56％～7.69％,外周血CDg7表达为5.21％～8.96％。30例急性髓细胞性白血病外周血及骨髓CD87表达差异有统计学意义（t=3．13,P〈0．05）。结论AML患者外周血及骨髓CD87表达有变化,外周血CD。表达高于骨髓,推荐应用外周血代替骨髓检测CD87表达。     

基因扫描分析CML病人外周血的T细胞克隆性

 应用RT-PCR方法扩增12例CML病人外周血单个核细胞的T细胞受体(CR)Vβ24个亚家族基因的互补决定区3(CDR3), 了解VβT细胞的分布情况, 8例正常人和T细胞株K37作为对照。 PCR产物进一步经基因扫描分析确定T细胞克隆性。 结果发现12例CML外周血中分别表达3－21Vβ亚家族T细胞, 8例正常人则表达除Vβ20外的全部VβT细胞, K37细胞仅表达Vβ9。基因扫描分析显示12例CML中8例病人的部分Vβ亚家族PCR产物至一主峰图象, 提示为克隆性生长的T细胞。 推测这是宿主对白血病细胞相关抗原的一种直接反应。 该方法有可能作为白血病特异性免疫治疗的临床研究的一种新的分子生物学研究方法。     

初治及复发急性髓细胞白血病患者BCRP基因表达的意义

目的 :探讨急性髓细胞白血病 (AML)患者初治及复发时乳腺癌耐药蛋白 (BCRP)基因表达与临床耐药及预后的关系。方法 :应用半定量逆转录—聚合酶链反应 (RT PCR) ,以 β2 微球蛋白 ( β2 MG)作内对照 ,检测了 5 1例初治及 1年内复发的 13例AML患者BCRP基因的表达 ,并检测了 15例正常人BCRP基因表达水平 ,将BCRP/ β2 MG≥ 0 4 6定为BCRP阳性。结果 :初治AML患者BCRP阳性率为 39 2 % ,BCRP阴性与BCRP阳性患者的首次完全缓解 (CR1)率分别为 90 3%和 35 0 %(P =0 0 0 0 0 ) ;13例一年内复发的患者BCRP基因表达水平 ( 1 0 6 5± 0 2 2 6 )显著高于初治时表达水平 ( 0 336± 0 2 0 8) (P =0 0 0 0 0 ) ;BCRP基因表达水平在AML的各亚型间无统计学差异。结论 :BCRP基因高表达可以导致临床耐药 ,是AML患者不利的预后因素。     

T细胞免疫球蛋白黏液素3基因在急性白血病中的表达及其临床意义

目的 探讨T细胞免疫球蛋白黏液素3(Tim-3)基因在急性白血病患者和白血病细胞株中的表达情况,并分析其临床意义.方法 收集初治急性白血病患者65例和8株白血病细胞株,同时收集15名健康供体的骨髓作为健康对照,分离单个核细胞,提取DNA,应用实时荧光定量聚合酶链反应检测单个核细胞上Tim-3 mRNA相对表达量,比较不同类型白血病Tim-3 mRNA表达差异,分析急性白血病患者Tim-3 mRNA表达与临床特征的关系.结果 M3细胞株NB4及非M3细胞株Kasumi-1、HL60、SHI-1、THP-1、Jurkat、CCRF-CEM、Mutz-1 Tim-3 mRNA相对表达量分别为(4.51±0.62)×10-5、(79.22±2.91)×10-5、(41.14±3.01)×10-5、(27.70±3.50)×10-5、(60.47±4.97)×10-5、(10.44±1.77)×10-5、(42.80±2.95)×10-5、(11.68±1.96)×10-5.非M3细胞株Tim-3 mRNA表达水平均高于NB4细胞株(均P＜0.05).非M3型急性髓系白血病(AML)患者Tim-3 mRNA相对表达量为(27.64±13.35)×10-4,AML-M3患者为(4.81±2.30)×10-4,急性淋巴细胞白血病(ALL)为(32.09±23.42)×10-4.其在非M3型AML和ALL中的表达水平高于健康对照[(12.02±4.22)×10-4],差异有统计学意义(P＜0.05),非M3型AML和ALL患者的Tim-3 mRNA表达水平差异无统计学意义(P＞0.05).在非M3型AML中,根据法、美、英协作组(FAB)分型,Tim-3 mRNA在AML-M4中的表达水平高于其他类型AML(P＜0.05).不同性别、年龄、骨髓原始细胞数、初诊白细胞数的急性白血病患者间Tim-3 mRNA表达水平差异均无统计学意义(均P＞0.05).结论 Tim-3 mRNA高表达于非M3型的急性白血病患者和细胞株,其表达水平与初治急性白血病患者的性别、年龄和原始细胞数无关.     

骨髓增生异常综合征bcl-2蛋白表达与细胞凋亡及增殖的关系

目的 :研究骨髓增生异常综合征 (MDS)中 bcl- 2蛋白表达与细胞凋亡及增殖的关系。方法 :以 2 0例 MDS患者、4例急性髓细胞白血病 (AML )患者、4例正常人的骨髓组织为研究对象 ,用免疫组织化学技术 SABC法分别检测 bcl- 2蛋白和增殖细胞核抗原 (PCNA )的表达 ,用 d U TP缺口末端标记 (TU NEL )技术检测细胞凋亡。结果 :MDS、AML、正常人 bcl- 2蛋白表达半定量积分值分别为 2 5 .35± 16 .6 8、6 6 .31± 10 .36、8.2 2± 2 .45 ;MDS、AML与正常对照组凋亡率依次递减 ;AML、MDS与正常对照组 PCNA阳性率依次递减 ;bcl- 2蛋白表达与凋亡负相关 (r=- 0 .6 76 1,P<0 .0 5 )。结论 :bcl- 2蛋白过度表达与细胞凋亡抑制、增殖失控关系密切 ,在 MDS向 AML转化过程中起重要作用     

Flt3基因在白血病中的表达及临床意义

目的 研究Ｆｌｔ３基因在不同类型、亚型白血病中ｍＲＮＡ表达、分布情况,以期阐明Ｆｌｔ３基因表达与疾病发生、发展的关系,探索Ｆｌｔ３基因作为白血病转归及预后指标的意义。方法 应用血细胞Ｐｅｒｃｏｌｌ不连续密度度分离法分离出外周血中原始细胞,采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）,检测Ｆｌｔ３基因在１０９例恶性血液病患者外周血中表达。结果 初治ＡＭＬ（急性髓细胞性白血病）中Ｆｌｔ３基因ｍＲＮＡ表达为     

Flt3 in acute myelogenous leukemia

Flt3 is a tyrosine kinase receptor expressed on hematopoietic cells. Activating mutations of this receptor are encountered in over one-fourth of acute myelogenous leukemia (AML) cases and activate multiple intracellular pathways leading to cell proliferation, inhibition of apoptosis, and blockage of differentiation in leukemic blasts. AML with flt3 mutations has a worse prognosis than AML with normal flt3, at least in younger patients. Sevaral flt3 inhibitors are in various stages of preclinical and clinical development and it is hoped that specific therapies against AML with flt3 mutations will soon be available to the clinician.     

Flt3 in acute myelogenous leukemia: biology, prognosis, and therapeutic implications

Flt3 is a tyrosine kinase receptor expressed on hematopoietic cells. Activating mutations of this receptor are encountered in over one-fourth of acute myelogenous leukemia (AML) cases and activate multiple intracellular pathways leading to cell proliferation, inhibition of apoptosis, and blockage of differentiation in leukemic blasts. AML with flt3 mutations has a worse prognosis than AML with normal flt3, at least in younger patients. Several flt3 inhibitors are in various stages of preclinical and clinical development and it is hoped that specific therapies against AML with flt3 mutations will soon be available to the clinician.     

急性髓系白血病中SMC3基因的表达及意义

目的:探讨急性髓系白血病（acute myeloid leukemia,AML）患者中SMC3基因表达的临床意义。方法:收集本院急性髓系白血病患者49例、缺铁性贫血（iron deficiency anemia,IDA）患者20例。应用实时荧光定量PCR(real-time quantitative PCR,qRT-PCR)法检测骨髓细胞中SMC3(structural maintenance of chromosome 3)mRNA表达,将患者分为高表达组和低表达组,分析SMC3表达水平与患者临床特征和预后分组之间的关系。结果:49例AML患者骨髓细胞标本SMC3 mRNA相对表达量高于对照组,SMC3基因表达与ELN（European Leukemia Net）AML遗传学预后分组相关（P=0.011）,高表达组中位生存时间4个月,低表达组中位生存时间为19个月,结果具有统计学显著性差异（P=0.000）。SMC3表达与年龄、性别、FAB分型、骨髓原始细胞、白细胞、血红蛋白、血小板计数均无统计学差异(P>0.05)。结论:SMC3 mRNA高表达与AML不良预后相关,作为可供筛选候补预后指标,其预后价值有待于在大样本研究中进一步验证。     

Inverse correlation between Flt3 and PU.1 expression in acute myeloblastic leukemias

Over-expression of the Flt3 is prevalent in acute myeloblastic leukemia (AML), playing a role in leukemogenesis while decreased expression of PU.1 induces AML in mice model. Therefore, we speculated that there is an inverse relationship between these two factors. To clarify this, we measured the expression level of Flt3 and PU.1 in 24 primary AML specimens. As a result, there is a significant negative correlation between Flt3 and PU.1 (r=-0.43, p<0.05). Furthermore, we revealed that flt3 gene promoter is suppressed by the over-expression of PU.1, suggesting that PU.1 is a potential suppressor of flt3 gene promoter.     

活化诱导胞嘧啶核苷脱氨酶基因在白血病中的表达

目的：探讨活化诱导胞嘧啶核苷脱氨酶（AID）在急性白血病（AL）、慢性粒细胞白血病慢性期（CML-CP）和急变期（CML-BC）患者及白血病细胞株中的表达特点。方法采用实时定量反转录聚合酶链反应（RT-PCR）检测89例初诊急性淋巴细胞白血病（ALL）患者、79例初诊急性髓系白血病（AML）患者、5例 CML-BC 患者、5例 CML-CP 患者和白血病细胞株 NB4、THP-1、KG-1、Raji、K562中 AID mRNA 的表达,以16名健康供者骨髓单个核细胞作为对照。结果89例 ALL 患者和79例 AML 患者 AID mRNA的中位表达水平分别为3.785（0.006～7463.175）、1.812（0.005～69.107）,健康对照组 AID mRNA 的表达水平为1.483（0.146～4.707）。 ALL、B-ALL、伯基特型白血病、Raji 细胞株和 M4的 AID mRNA 表达水平较健康对照组升高,差异均有统计学意义（均 P＜0.05）,而 M3、NB4和 KG-1细胞株的 AID mRNA 表达水平较健康对照组下降,差异均有统计学意义（均 P＜0.05）。 K562细胞株的 AID mRNA 表达水平较 CML-CP患者升高,差异有统计学意义（P＜0.001）。 CML-BC、CML 淋系急性变（CML-LBC）和 CML 髓系急性变（CML-MBC）患者的 AID mRNA 表达水平也高于 CML-CP 患者。结论 AID 在 B-ALL、伯基特型白血病、M4中呈高水平,在 M3和 KG-1细胞株中呈低表达,在 CML-BC 中出现表达水平的显著升高。     

Endogenous vascular endothelial growth factor-C expression is associated with decreased drug responsiveness in childhood acute myeloid leukemia.

PURPOSE: We hypothesized that downstream effects of endogenous vascular endothelial growth factor (VEGF)/VEGF receptor signaling on acute myelogenous leukemia (AML) cell survival resulted in increased in vitro cellular drug resistance and a longer time to kill most leukemic cells in vivo upon drug exposure. EXPERIMENTAL DESIGN: In primary AML cells from pediatric patients, VEGFA and VEGFC mRNA expression and in vitro cellular resistance to nine cytotoxic drugs were studied. As in vivo equivalents for in vitro drug resistance, in vivo AML blast reduction upon drug exposure, measured as blast cell reduction on day 15 in the bone marrow and as time in days from diagnosis to complete remission (CR) were used. RESULTS: Increased endogenous VEGFC levels significantly correlated with increased in vitro resistance for six typical AML drugs in primary AML cells from pediatric patients. Patients with >5% blasts on day 15 showed a 12.9-fold increase in the median VEGFC level compared with patients with < or =5% blasts (P = 0.002). Time to reach CR was studied using linear regression analysis with VEGFC, age at diagnosis, sex, treatment protocol, FAB type, cytogenetic risk profile, and WBC counts as variables. There was a significant positive independent association between VEGFC levels and time to CR (b = 6.02, SE = 1.58, P < or = 0.0001, n = 72). CONCLUSIONS: These results suggest for the first time that higher endogenous VEGFC levels of AML cells are related to decreased in vitro and in vivo drug responsiveness.     

Folate receptor type beta is a neutrophilic lineage marker and is differentially expressed in myeloid leukemia

BACKGROUND: The membrane-associated folate receptor (FR) type beta is elevated in the spleen in patients with chronic myeloid leukemia (CML) and acute myeloid leukemia (AML). In this study, the authors investigated possible cell type and differentiation stage specificity of expression of FR-beta in normal and leukemic hematopoietic cells. METHODS: An affinity-purified rabbit polyclonal antibody specific for FR-beta was employed for immunostaining representative bone marrow smears and peripheral blood smears from normal individuals and from a limited number of patients with various leukemias. Multiple samples of normal bone marrow and peripheral blood were analyzed for the expression of FR-beta and selected CD antigens by two- or three-color flow cytometry. RESULTS: Of the morphologically identifiable cells, only neutrophils were positive for FR-beta. The leukemic blasts in CML patients showed expression of FR-beta with no apparent relation to the occurrence of the Philadelphia chromosome. Among acute nonlymphocytic leukemias, FR-beta was expressed in promyelocytic leukemia, in the myeloblast populations of myelomonocytic and erythroleukemias, and variably in M1/M2 AML. Neither the blasts of acute lymphocytic leukemia nor the more mature cells of chronic lymphocytic and hairy cell leukemias expressed FR-beta. The less differentiated FR-beta positive AML samples also were positive for CD34 and HLA-DR. Flow cytometric analysis of normal bone marrow and peripheral blood revealed low or insignificant coexpression of FR-beta with CD34, CD19, and CD3, whereas significant coexpression was observed with high levels of CD33, CD13, and CD11b; coexpression of FR-beta with CD14 was high in the immature bone marrow cells, comparable to that in myeloid cells, but relatively low in peripheral blood. CONCLUSIONS: The results of this study suggest a narrow expression pattern of FR-beta marking the neutrophilic lineage and the possibility of defining a subtype or subtypes of myeloid leukemia based on FR-beta expression. The identification of FR-beta positive leukemias and the absence of the receptor in normal CD34 positive cells may enable selective receptor-mediated targeting of leukemic cells.     

SAMHD1基因在初诊急性髓细胞白血病患者中的表达及其临床意义

目的:探讨SAMHD1基因在初诊急性髓细胞白血病（AML)患者中的表达及其临床意义。方法:实时定量PCR方法检测13例成人初诊AML患者与7例非恶性血液病患者SAMHD1 mRNA的表达,分析其临床意义。利用SPSS统计学方法分析SAMHD1基因表达高低与初诊白血病患者临床和实验室特征之间的相关性,采用Kaplan-Meier法以及COX回归模型进行多因素生存分析。结果:SAMHD1基因在AML患者中的表达低于非恶性血液病患者（P＜0.05）,且SAMHD1基因表达高低与患者性别、年龄、初诊时WBC、HGB、PLT、骨髓原始细胞百分比分布差异均无统计学意义;SAMHD1表达高低在预后良好与预后不良组间差异比较有统计学意义（P＜0.05),其中低表达组的OS低于高表达组。结论:SAMHD1基因在AML与非恶性血液病患者中表达差异具有统计学意义,且其可作为影响初诊AML患者OS的独立预后因素。