血液肿瘤患者FLT3/ITD基因突变的检测及临床意义

目的检测血液肿瘤患者FLT3/ITD基因突变,探讨其突变的临床意义。方法2001—2005年对南方医科大学南方医院血液科332例血液肿瘤患者,采用聚合酶链反应(PCR)方法检测FLT3/ITD基因突变。结果FLT3/ITD基因突变阳性率分别为急性髓性白血病(AML)22.3%(23/103)、慢性髓性白血病急变期(CML-BC)6.5%(2/31)、骨髓增生异常综合征(MDS)5.6%(2/36)和急性淋巴细胞白血病(ALL)2.6%(2/76)。而慢性髓性白血病慢性期(CML-CP)、慢性淋巴细胞白血病(CLL)、多发性骨髓瘤(MM)和非霍奇金淋巴瘤(NHL)均未发现FLT3/ITD基因突变。FLT3/ITD基因突变阳性AML患者外周血WBC计数及骨髓白血病细胞比例显著高于FLT3/ITD基因突变阴性AML患者(P<0.05),FLT3/ITD基因突变阳性AML患者完全缓解后18个月内累计复发率(63.6%)显著高于FLT3/ITD基因突变阴性AML组(27.7%)(P<0.05)。结论FLT3/ITD基因突变检测对血液肿瘤预后有一定意义;FLT3/ITD基因突变AML患者预后差。     

急性淋巴细胞白血病细胞克隆性分析的临床意义

采用多聚酶链反应 (polymerase chain reaction,PCR)技术检测 49例急性淋巴细胞白血病患者免疫球蛋白重链 (imm unoglobulin heavy chain,Ig H)基因重排 ,并分析其细胞克隆性。结果 :35例显示有克隆性 Ig H重排 ,其中单克隆 2 6例 ,寡克隆 9例。9例完全缓解期中多次动态监测表明 :PCR检测 Ig H重排 ,在微小残留病监测和疾病复发判断中 ,具有重要的临床意义。     

FLT3在急性髓细胞白血病中的研究进展

急性髓细胞白血病（AML）是一组具有不同生物学特性的恶性血液肿瘤。近年来,分子遗传学研究的进展使得大多数AML可以检出基因水平的异常,其中FLT3基因突变在AML发生及预后中起重要作用,并为白血病的靶向治疗提供了新的目标。本文对此作一综述。     

PCR方法检测急性髓系白血病FLT3的表达

近年来,在鉴定急性髓系白血病(AML)获得性分子标志方面已有很大进步,而且这可能对现在使用的依据细胞遗传学进行的危险分层增加一些新的信息,特别是为成年人核型正常的AML的治疗及预后评估提供新的借鉴。国外自2000年以来已开展了这方面的实验室检测及临床工作,我国则刚刚起步。现将本人自2008年以来做的关于     

应用PCR检测急性淋巴细胞性白血病患者脑脊液中白血病细胞

应用ＰＣＲ检测急性淋巴细胞性白血病患者脑脊液中白血病细胞徐兵周淑芸张剑免疫球蛋白重链（ＩｇＨ）和Ｔ细胞受体（ＴＣＲ）基因重排可作为淋巴系肿瘤特异性基因标志。我们采用聚合酶链反应（ＰＣＲ）技术检测急性淋巴细胞性白血病（急淋）患者脑脊液（ＣＳＦ）标本Ｉｇ...     

急性淋巴细胞白血病免疫球蛋白重链及T细胞受体γ基因重排的临床观察

免疫球蛋白重链 (ＩｇＨ )基因和Ｔ细胞受体 (ＴＣＲ)ｒ基因的重排可作为淋巴细胞白血病的特异性基因标志。应用敏感、特异的多聚酶链反应 (ＰＣＲ)技术检测ＩｇＨ及ＴＣＲγ基因重排 ,对急性淋巴细胞白血病 (ＡＬＬ)的基因分型、微小残留病变 (ＭＲＤ)的检测、判断疗效、预测复发等均具有重要意义。我们于 1997年～ 1998年应用ＰＣＲ技术对 42例成人ＡＬＬ病人的ＩｇＨ及ＴＣＲγ的基因重排进行研究。对象与方法1.对象 :ＡＬＬ患者 42例 ,男性 2 5例 ,女性 17例。年龄 15～ 70岁 ,中位年龄 3 5岁。分型 :Ｌ16例 ,Ｌ2 2 8例 ,Ｌ3 8例。其中…     

FLT3突变急性髓系白血病的全程管理

约30%的新诊断急性髓系白血病(AML)患者携带FMS样受体酪氨酸激酶3(FLT3)基因突变，大多数FLT3突变是近膜结构域内的内部串联重复(ITD),也有少数是酪氨酸激酶结构域(TKD)内的点突变。FLT3抑制剂的应用改变了FLT3突变AML患者的治疗现状与预后。文章就FLT3突变AML的生物学特征，FLT3突变检测与诊断、FLT3抑制剂的应用及异基因造血干细胞移植中的作用等全程管理进行讨论。     

急性髓细胞白血病FLT3基因突变及靶向治疗研究进展

急性髓细胞白血病（acute myeloid leukemia，AML）是一组起源于造血干细胞的恶性克隆性疾病，具有高度异质性。由基因突变导致的细胞增殖、分化及凋亡途径改变是AML的发病基础。近年来随着分子生物学技术的发展和对AML发病机制的深入研究，发现了一些与AML发生发展密切相关的基因异常，为AML的个体化治疗和预后判断提供了新的依据，某些还成为潜在的新治疗靶点。FLT3（FMS—like receptor tyrosine kinase），又称胎肝激酶2（fetal liver kinase2，FLK2）或干细胞酪氨酸激酶1（stem cell tyrosine kinase-1，STK-1），与FMS、KIT、血小板衍生生长因子（PDGFR）等同属于Ⅲ型受体酪氨酸激酶家族（RTK）。该基因及其配体在干细胞的分化、增殖中发挥重要作用。FLT3基因突变在AML中发生率较高，对预后影响较大，目前对该基因的研究正逐步深入，现综述如下。     

银染PCR—SSCP方法检测61例白血病P53基因点突变

用银染PCR－SSCP方法检测61例不同类型的白血病P53抗癌基因点突变，结果在26例淋巴细胞白血病中检测出2例有P53基因的点突变，在35例髓系白血病中检测出1例有P53基因的点突变。临床观察3例P53基因点突变患者对化疗药物不敏感，其中2例病程进展凶险。结果显示，P53抗癌基因点突变导致功能失活在白血病发病过程中起一定的作用，并可作为判断预后和化疗反应性的检测指标。     

复发/难治性急性淋巴细胞白血病患者多药耐药基因表达及临床意义

目的探讨复发/难治性急性淋巴细胞白血病(ALL)患者多药耐药基因(mdr1基因)的表达及其临床意义。方法采用Northern blot法检测K562/A02细胞株mdr1基因的表达,RT-PCR法检测42例ALL患者(ALL组)、27例非白血病患者和健康人(对照组)mdr1基因的表达水平,分析mdr1基因过度表达的临床意义。结果ALL组mdr1基因过度表达率显著高于对照组(P<0.01),其中复发/难治者高于初诊和完全缓解(CR)者(P<0.01),临床耐药者高于非临床耐药者(P<0.01);非临床耐药者CR率显著高于临床耐药者(P<0.001)。结论mdr1基因过度表达与ALL患者临床耐药密切相关,是影响预后的重要因素;mdr1基因表达可作为判断ALL患者临床耐药和预后的指标。     

Rapid detection of chimeric bcr/abl mRNAs in acute lymphoblastic and chronic myeloid leukemia by the polymerase chain reaction

Summary We have developed a rapid method for the detection of bcr/abl mRNAs, the products of the BCR/ABL fusion genes. The method is based on the polymerasechain-reaction (PCR). Through the use of additional internal primers it is possible to detect directly a single Ph¹-positive cell among 10⁵ unaffected cells thus omitting time-consuming blotting procedures. The whole analytical procedure starting from RNA isolation to agarose gel electrophoresis including two rounds of PCR can be performed in less than six hours.Supported by the Wilhelm Sander-Stiftung, Neustadt/Donau and the Deutsche Krebsgesellschaft, Landesverband Berlin     

Salvage therapy using FLT3 inhibitors may improve long‐term outcome of relapsed or refractory AML in patients with FLT3‐ITD

To determine the long‐term efficacy of FLT3 inhibitors (FLT3i) in the salvage setting for relapsed and refractory (rel/ref) acute myeloid leukemia (AML) with FLT3 internal tandem duplication (AML FLT3‐ITD), we conducted a retrospective study of 120 patients with rel/ref AML FLT3‐ITD who received salvage therapy with either FLT3i‐containing regimen (FLT3i group, N = 45) or conventional cytotoxic regimen (conventional group, N = 75). The median overall survival (OS) after the first salvage in the FLT3i group was 6·9 vs. 4·6 months in the conventional group (P = 0·17). The OS was better in the FLT3i group among patients with initial complete remission (CR) duration ≤12 months or with primary refractory disease (6·9 vs. 3·7 months; P < 0·01). The OS was better when FLT3i was combined with cytotoxic agents versus monotherapy (17 vs. 4·8 months; P = 0·017). Multivariate analysis revealed that the use of FLT3i was an independent predictor of OS (hazard ratio 0·58; 95% confidence interval, 0·38–0·88). Incorporating FLT3i into salvage strategies may improve long‐term outcome of patients with AML FLT3‐ITD. Prospective studies to validate this conclusion are warranted.     

The clinical significance of FLT3 ITD mutation on the prognosis of adult acute promyelocytic leukemia

Background and aims: To explore the relationship between FLT3 (encoding Fms related tyrosine kinase 3) internal tandem duplication (ITD) mutations with the prognosis of acute promyelocytic leukemia. The PubMed database, the Cochrane Library, conference proceedings, the EMBASE databases, and references of published trials and review articles were searched. Two reviewers independently assessed the quality of the trials and extracted the data. Odd ratios (ORs) for complete remission (CR) rate after induction therapy, 5-year overall survival (OS), and 5-year disease free survival (DFS) were pooled using the STATA package.

Main results: Seventeen trials involving 2252 patients were ultimately analyzed. The pooled OR showed that the FLT3 ITD mutation group had a poor prognosis in terms of CR rate (OR?=?0.53, 95% confidence interval (CI), 0.30–0.95, P?=?0.03), 5-year OS (OR?=?0.47, 95% CI, 0.29–0.75, P?=?0.002), and as 5-year DFS (OR?=?0.48, 95% CI, 0.29–0.78; p?=?0.003).

Conclusions: The results suggested that FLT3 ITD mutations could become an indicator of poor prognosis of APL, and these patients should receive more intensive therapy according to current guidelines.     

Sorafenib plus all‐trans retinoic acid for AML patients with FLT3‐ITD and NPM1 mutations

Knowledge of the molecular basis of acute myeloid leukaemia has increased considerably in the past few years, and therapies targeting specific molecular defects of this disease are intensively investigated. Patients with both NPM1 and FLT3‐ITD mutations encompass 20% of cytogenetically normal AML. The multikinase and FLT3 inhibitor, sorafenib, has shown some efficacy in patients with relapsed FLT3‐ITD⁺ AML. In addition, it is suggested that all‐trans retinoic acid (ATRA) used in combination with chemotherapy has shown to improve outcome of patients harbouring NPM1 mutations. We report here the clinical course of three patients with refractory or relapsed FLT3‐ITD⁺/NPM1⁺ AML who achieved significant response upon sorafenib and ATRA combination.     

ADAMTSL5 and CDH11: putative epigenetic markers for therapeutic resistance in acute lymphoblastic leukemia

Background and objectives: DNA hypermethylation has been linked to poor treatment outcome in childhood acute lymphoblastic leukemia (ALL). Genes differentially methylated in the chemoresponsive pre-B-ALL compared to chemoresistant pre-B-ALL cases provide potential prognostic markers.

Methods: DNA methylation profiles of five B-ALL childhood patients who achieved morphological complete remission (chemoresponsive) and five B-ALL patients who did not (chemoresistant) after induction treatments as well as four normal controls were compared on 27?000 CpG sites microarray chips. Subsequently, methylation-specific polymerase chain reaction (MSP) on selected hypermethylated genes was conducted on an additional 37 chemoresponsive and 9 chemoresistant B-ALL samples and 2 normal controls.

Results: Both methods were found to be highly correlated. Unsupervised principal component analysis showed that the chemotherapy-responsive and -resistant B-ALL patients could be segregated from one another. Selection of segregated genes at high stringency identified two potential genes (CDH11 and ADAMTSL5). MSP analysis on the larger cohort of samples (42 chemoresponsive, 14 chemoresistant B-ALL samples and 6 normal controls) revealed significantly higher rates of hypermethylation in chemoresistant samples for ADAMTSL5 (93 vs. 38%; p?=?0.0001) and CDH11 (79% vs. 40%, p?Conclusion: Chemoresistant B-ALL patients are associated with increased methylation in ADAMTSL5 and CDH11. These findings need to be validated in a larger group of patients, and the functional biological and prognostic significance of differential methylation needs to be studied further.     

中国儿童急性淋巴细胞白血病GATA3基因单核苷酸多态性与预后的相关性研究

目的 研究中国儿童急性淋巴细胞白血病(ALL)患儿GATA3基因单核苷酸多态性(SNP)分布特点及其对预后的影响.方法 利用TaqMan探针荧光定量PCR法检测上海儿童医学中心2009-05～2014-12收治的678例ALL患者GATA3基因rs3781093和rs3824662两个SNP位点,分析其与早期治疗反应、...