BCL6、KLF5、NCL基因在儿童急性淋巴细胞白血病中异常表达的特点

目的:研究转录因子基因BCL6、KLF5及核仁蛋白基因NCL在急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)患儿骨髓细胞中的表达情况及其在不同疾病状态下的表达特点。方法:选取北京儿童医院2004年1月至2005年12月住院ALL患儿100例,另取由于骨骼畸形而在北京儿童医院进行外科手术的5例非ALL患儿作对照;以基因芯片检测ALL患儿骨髓细胞中异常表达基因,GeXP多重基因表达分析系统检测BCL6、KLF5、NCL基因在另外选取的10例配对ALL患儿初诊及缓解期的表达变化。结果:基因芯片筛查发现,在100例各亚型ALL标本中,BCL6和KLF5mRNA表达均下调,NCLmRNA表达均上调。BCL6和KLF5mRNA在10例ALL初诊患儿骨髓细胞中表达较低,完全缓解后表达升高(0.380±0.16vs0.850±0.10,0.074±0.021vs0.228±0.049;均P<0.01);NCLmRNA在ALL初诊患儿骨髓细胞中表达较高,完全缓解后表达降低(0.234±0.054vs0.151±0.055,P<0.01)。在10例配对患者儿中,TEL-AML1阳性及E2A...     

儿童急性淋巴细胞白血病治疗现状

近20余年儿童急性淋巴细胞白血病（ALL）长期无病生存率（EFS）达70%-80%。长期ESF率的提高主要归功以下三方面工作：（1）由于分子生物学研究进展，更准确地判断预后，采用MIC或MICM分型，按型用化疗方案。（2）化疗方案合理组合在保证疗效的同时对ALL后期治疗降低强度，减少药物毒性，同时改进了庇护所预防措施-采用大剂蛳氨甲蝶呤（HD-MTX），大大减少了髓外复发，使相当一部分患儿放弃以往颅脑预防性放疗，即使需要放疗者也降低了剂量，从而减少放化疗远期毒副作用，提高了生存质量。（3）强有力的支持治疗如：空气净化层流病房的建立，有效抗菌素，抗病毒、抗霉菌药物高效价丙种球蛋白等的应用，预防治疗也骨髓抑制期的感染，造血因子、GM、CSF、G-CSF浓缩红细胞、血小板的输注缩短了强化疗所致骨髓抑制期。加速了造血的恢复。     

儿童急性淋巴细胞白血病免疫表型测定

急性淋巴细胞白血病 (ＡＬＬ)是儿童高发的恶性肿瘤之一。免疫表型 (免疫学 )在诊断该病中起着其它方法所不能替代的重要作用 ,如Ｔ ＡＬＬ和兼备淋、髓同时表达及伴ＣＤ3 4阳性的ＡＬＬ ,必须依靠免疫表型来诊断。作者在近 5年用免疫酶标法和 19种单克隆抗体 (单抗 )对 6 0例儿童ＡＬＬ进行了免疫表型的测定与分析 ,现报道如下。材料与方法一、对象　 6 0例为 1994年 8月～ 1999年 11月就诊的初发未经治疗的儿童ＡＬＬ ,男 37例 ,女 2 3例 ,年龄 2 0 8～ 13 6 7岁 ,平均 7 99岁。骨髓按ＦＡＢ分类Ｌ12 9例 ,Ｌ2 2 6例 ,Ｌ3 5例。同时 ,作…     

儿童pro-B急性淋巴细胞白血病的临床特点及预后分析

目的 探讨儿童pro-B急性淋巴细胞白血病(pro-B-ALL)的临床特点及预后。方法 选择ALL患儿696例,统计pro-B-ALL发生率,并分析其临床特点、治疗和预后。结果 ALL患儿中pro-B-ALL发生率为6.18%(43/696),其中男性24例,女性19例;年龄<1岁3例,1～9岁23例,≥10岁17例;初诊白细胞(WBC)计数≥50×10⁹/L 15例,<50×10⁹/L 28例;免疫表型中B系抗原CD20阳性5例、CD22阳性30例;T系抗原CD7阳性5例;干/祖细胞抗原CD34阳性31例,CD58阳性22例、CD15阳性5例;髓系抗原CD33阳性13例,CD13阳性10例;融合基因中混合谱系白血病基因重排(MLL-r)阳性15例,E2A-PBX1阳性1例,BCR-ABL1阳性1例,MLL-AF4阳性8例;MLL-r、MLL-AF4阳性者初诊WBC≥50×10⁹/L比例均较阴性者高,CD13、CD22阳性率均较阴性者低,差异有统计学意义(P<0.05)。诱导缓解治疗1个月后完全缓解(C...     

急性淋巴细胞白血病TPMT基因多态性的研究

目的 分析急性淋巴细胞白血病TPMT基因多态性与6-MP不良反应的相关性.方法 收取急性淋巴细胞白血病患儿48例作为研究对象,对其TPMT基因型及6-MP不良反应进行分析.结果 48例患儿中31.25％按照常规6-MP使用剂量完成维持治疗,68.75％患儿出现不耐受后调整为低剂量完成维持治疗.常规剂量组重度不良反应发生率高于低剂量组,但差异无统计学意义(P＞0.05).6-MP所致骨髓抑制及肝功能损害发生率分别为93.75％及83.33％.仅有1例患儿发生杂合型TPMT×3C点突变,突变发生率为2.08％,该患儿同时发生4级骨髓抑制及4级肝功能损害.结论 TPMT×3C基因突变可能与6-MP所致重度不良反应有关,但6-MP所致重度不良反应可能是多种因素共同作用的结果.     

成年人T细胞急性淋巴细胞白血病中Ikaros6表达及其临床意义

目的 检测成年T细胞急性淋巴细胞白血病（T-ALL）患者Ikaros6的表达,并探讨其临床意义.方法 提取成年T-ALL患者初发时骨髓单个核细胞的RNA,反转录为cDNA,采用反转录-聚合酶链反应（RT-PCR）扩增技术进行检测,测序进一步确定结果,并结合患者临床资料分析临床特点及其预后.结果 74例初发成年人T-ALL中,Ikaros6阳性率为21.6％（16例）,另外阳性患者较阴性患者更易发生髓外浸润（x2=4.084,P=0.043）,且外周血白细胞计数高（103.15×109/L比15.60×109/L）、贫血更严重（血红蛋白75.95 g/L比107.00 g/L）和血小板计数低（26.0×109/L比67.0×109/L）,但差异均无统计学意义（均P＞ 0.05）.Ikaros6阳性患者生存期短和具有高复发风险.结论 Ikaros6在成年人T-ALL中表达率较高,阳性患者具有外周血白细胞计数高和易出现髓外浸润等临床特点,预后差,具有高复发风险.Ikaros6可考虑作为成年人T-ALL分层治疗或预后判断的分子指标.     

血浆miRNAs在儿童急性淋巴细胞白血病(ALL)不同疾病状态中的表达特点

目的 分析3种血浆miRNA在儿童急性淋巴细胞白血病(ALL)不同疾病状态中的表达特点.方法 选取急性淋巴细胞白血病患儿40例,其中初诊患儿12例,完全缓解患儿16例,复发患儿12例,同时收集我院经健康体检的儿童25例作为正常对照组.抽取所有儿童的空腹静脉血,离心后取血浆,采取直接扩增目的microRNA,并反转录为cDNA,qRT-PCR方法检测血浆miR150、miR155、miR233的表达.结果 ALL组中血浆miR150、miR155、miR233的表达均与正常组有统计学差异;血浆miR150和miR233在初诊组和复发组的表达显著低于缓解组和正常组(P<0.05),血浆miR155在初诊组和复发组的表达显著高于缓解组和正常组(P<0.05).3种miRNA在初诊组与复发组之间、缓解组与正常组之间比较均无统计学差异(P>0.05);初诊组中3种血浆miRNA在不同年龄和性别之间表达均无统计学差异(P>0.05);3种血浆miRNA的AUC(曲线下面积)和95%CI(95%可信区间)均有较好的满意度,且3种血浆miRNA联合检测的精确度更高.结论 血浆miR150、miR155、miR233的表达在儿童急性淋巴细胞白血病(ALL)的诊断、治疗和复发监测中有较高的应用价值.     

儿童急性淋巴细胞白血病治疗现状

自70年代以来，随着对白血病细胞动力学认识的深化，新的治疗药物不断涌现及治疗方法的改进，国外，儿童急性淋巴细胞白血病(急淋)的完全缓解率(CR)已达90～100％，5年以上无病生存率已达70％以上。国内我院及北京儿童医院最近几年总结的材料表明，无论在CR率及5年以上无病生存率方面，均已达到国外所报道的水平，从而根本性地改变了急性白血病是“不治之症”的慨念。     

miR-128在急性淋巴细胞白血病中的表达

目的 探讨miR-128在急性淋巴细胞白血病(ALL)中的表达及其意义.方法 采用实时荧光定量聚合酶链反应法对62例ALL患者和20例骨髓正常患者骨髓单个核细胞miR-128的相对表达量进行检测,并与患者的临床资料进行相关性分析.结果 miR-128在ALL患者中的相对表达量高于健康对照组(P＜0.05),但其在急性B淋巴细胞白血病(B-ALL)及急性T淋巴细胞白血病(T-ALL)中的表达水平差异无统计学意义(P＞0.05),miR-128的表达与是否存在bcr-abl融合基因及其mRNA的表达水平均无相关性(均P＞ 0.05).结论 miR-128在ALL患者中表达上调,可作为临床诊断ALL的潜在分子标志物,bcr-abl融合基因的表达对miR-128在ALL中的表达无明显影响.     

儿童急性淋巴细胞白血病的研究

我院1978年-1988年收治的139例儿童急性淋巴细胞白血病(ALL)，其完全缓解(CR)率达97．1％，系统随访治疗的105例病人5年以上持续完全缓解(CCR)率是58．6％，5年以上生存率达71．9％，达到国际先进水平。现就本研究得出的关于儿童ALL长期无病生存的几个关键问题阐述如下。     

Osteonecrosis in pediatric patients with acute lymphoblastic leukemia

The authors report five pediatric patients with acute lymphoblastic leukemia (ALL) in whom symptomatic aseptic osteonecrosis developed on therapy. All patients had been on treatment with a modified BFM protocol and developed osteonecrosis in the maintenance phase of the protocol. The avascular necrosis was multifocal in all. The authors' data suggest that dexamethasone used in the reinduction phase of the protocol may be the responsible agent although no definite proof exists. Since only symptomatic patients are reported, the true frequency of this complication may be significantly higher.     

Aberrant expression of tumor suppressor genes and their association with chimeric oncogenes in pediatric acute lymphoblastic leukemia

Aberrant expression of tumor suppressor genes WT 1, RB 1, p53, homozygous deletion of p16 gene and their relationship with expression of oncogenes BCR-ABL, TEL-AML 1, MLL-AF 4, E2A-PBX 1, SIL-TAL 1 were determined in bone marrow samples of children with de novo B-lineage (n=170) and T-lineage (n=25) acute lymphoblastic leukemia (ALL). In contrast to expression of chimeric oncogenes alterations in p16, WT 1, RB 1 and p53 expression were T/B-lineage-unrestricted. Significant association between expression of MLL-AF 4 and WT 1, E2A-PBX 1 and p53; SIL-TAL 1 and homozygous deletion of p16 has been demonstrated.     

Urolithiasis in pediatric patients with acute lymphoblastic leukemia.

We evaluated the incidence, timing, and consequences of urolithiasis in children with acute lymphoblastic leukemia (ALL). A total of 20 patients with urolithiasis were identified from 2095 patients with ALL treated at St Jude Children's Research Hospital on consecutive protocols between 1968 and 1998. For remission induction therapy, all patients received daily prednisone; continuation chemotherapy regimens differed by protocol with some including pulses of prednisone or dexamethasone and others no glucocorticoid. Patients with urolithiasis were older at diagnosis of ALL than those without urolithiasis (median age, 7.5 vs 5.0 years; P=0.03) and less likely to be black (P=0.03) than white or Hispanic, but sex and treatment era did not differ. Presenting symptoms included abdominal or flank pain, hematuria, and dysuria. All stones analyzed biochemically were calcium stones. The incidence of urolithiasis after completion of therapy was 1.8 per 10 000 person-years. Compared to this baseline rate, the relative risk of urolithiasis was 45 (P<0.01) during induction therapy, 22 (P<0.01) during continuation therapy with glucocorticoids, and 5.1 (P>0.05) during continuation therapy without glucocorticoids. Urolithiasis occurred 4.5 times more often during continuation treatment with glucocorticoids than without (P<0.05). Seven patients (35%) had recurrent urolithiasis. Patients with ALL are at risk of developing calcium renal stones during chemotherapy, especially when a glucocorticoid is included.     

Aberrant DNA methylation in pediatric patients with acute lymphocytic leukemia

BACKGROUND: Aberrant methylation of promoter-associated cystosine-guanine (CpG) islands is an epigenetic modification of DNA frequently observed in adult patients with acute lymphocytic leukemia (ALL). This epigenetic modification has been associated with gene silencing, malignant transformation, and aging. It is not known whether there are epigenetic differences between pediatric patients and adult patients with ALL. METHODS: To investigate the methylation characteristics of pediatric patients with ALL and to determine whether DNA methylation can explain prognostic or biologic differences between pediatric and adult patients, the authors analyzed the methylation status of 7 promoter-associated CpG islands in 16 pediatric patients with ALL and compared them with the methylation characteristics of a cohort of adult patients with ALL. The genes analyzed included the estrogen receptor gene (ER), multidrug resistance gene 1 (MDR1), p15, C-ABL, CD10, p16, and p73. RESULTS: The mean methylation densities of ER, MDR1, CD10, p15, and C-ABL were 25.4%, 16.4%, 5.23%, 4.24%, and 4%, respectively. P16 was methylated in 11.7% of patients, and p73 was methylated in 17.6% of patients. One patient (6.2%) had methylation of 0 genes, 15 patients (93.7%) had methylation of >/= 1 gene, and 4 patients (25%) had methylation of 3-4 genes. Methylation of all these genes was < 2% (or methylation specific polymerase chain reaction negative) in nonneoplastic tissues. A significant inverse correlation was observed between methylation of CD10 and CD10 expression. No differences were observed between the methylation characteristics of pediatric patients and adult patients. CONCLUSIONS: The results indicate that DNA methylation is common in pediatric patients with ALL and that methylation of the genes studied does not account for prognostic differences between pediatric patients and adult patients with ALL.     

HLA-DM expression is elevated in ETV6-AML1 translocation-positive pediatric acute lymphoblastic leukemia

MHC Class II antigen processing and presentation to CD4+ T cells is important in anti-tumor immunity. ETV6-AML1-positive precursor-B acute lymphoblastic leukemia (pre-B ALL) is associated with good outcome and late relapse. We analyzed HLA-DR and the MHC Class II processing components HLA-DO, HLA-DM, and Class II-associated invariant chain peptide (CLIP) expression, in ETV6-AML1-positive and ETV6-AML1-negative ALL. Overall, CLIP expression was low, while HLA-DM was significantly higher in ETV6-AML1-positive cells. No significant difference in HLA-DO was observed. The HLA-DR(high)/CLIP(low)/HLA-DM(high) phenotype strongly suggests that ETV6-AML1 leukemia will induce favorable immune responses and may in part explain the characteristic late relapses associated with ETV6-AML1 pre-B ALL.     

Dexamethasone alters sleep and fatigue in pediatric patients with acute lymphoblastic leukemia

BACKGROUND: Dexamethasone improves the cure rate of childhood acute lymphoblastic leukemia (ALL) but causes physical and behavioral adverse events. The objective of the current study was to determine the effect of dexamethasone exposure on sleep and fatigue in pediatric patients with ALL. METHODS: One hundred pediatric patients with low-risk or standard-risk ALL were enrolled on 1 of 3 protocols (St. Jude Total XV, Children's Oncology Group [COG] 9904, or COG 9905) at 3 institutions. The mean age of the cohort was 9.24 +/- 3.23 years (range, 5.03-18.14 years). The majority of patients were white (79%) males (62%) with standard-risk ALL (63%). The cohort was divided into 4 subgroups: St. Jude low-risk, St. Jude standard-risk, COG low-risk, and COG standard-risk. Patients wore a wrist actigraph to monitor sleep activity during 2 consecutive 5-day periods: During the first period, they did not receive dexamethasone; and, during the second period, they did. Patients and their parents completed fatigue instruments on Days 2 and 5 of each period, and parents completed sleep diaries. RESULTS: Actual sleep minutes, sleep duration, total daily nap minutes, and fatigue increased significantly during the dexamethasone treatment for 3 to 4 of the subgroups. Total daily nap minutes increased significantly for both standard-risk groups during the dexamethasone treatment. Parents reported significant increases in their child's nighttime awakenings, restless sleep, and nap time during dexamethasone treatment. CONCLUSIONS: Dexamethasone treatment during continuation therapy for childhood ALL significantly and adversely altered sleep and fatigue, confirming that sleep and fatigue are behavioral responses to dexamethasone.     

Jak3 expression and genomic sequence in pediatric acute lymphoblastic leukemia

Janus tyrosine kinase 3 (JAK3) is one of several key regulatory enzymes in B-cell precursors which is highly conserved between multiple species. The gene for Jak3 has been mapped to human chromosome 19p12-13.1 and encompasses 23 exons. Constitutively high levels of JAK3 activity may contribute to drug resistance and enhanced clonogenicity of leukemic B-cell precursors from children and infants with acute lymphoblastic leukemia (ALL). As part of a systematic effort to accurately determine the genomic sequence of Jak3 gene in normal and leukemic B-cell precursors, we sequenced a relatively short region of Jak3 spanning two introns, originally termed introns 10 and 11. This genomic sequence appeared in certain RT-PCR products from our analysis of Jak3 gene expression in pediatric, as well as infant, primary ALL cells. Unexpectedly, a gap in the original Jak3 genomic sequence was found in intron 10 across the sequence matching to an Alu element. Furthermore, the sequence obtained from intron 11 did not match at all to that previously reported, and the length of the intron was much larger than expected at 1.1 kb. Homology to Alu elements (three regions, 699 bp total) and a LINE2 element (one region, 189 bp total) were seen across the entire region covering exons 10-12 (2.1 kb total). Two potential single nucleotide polymorphisms (SNPs) were observed in intron 11. No apparent genomic mutation was found across this region in leukemic B-cell precursors from any of the ALL patients examined. This newly described sequence corrects the previous published genomic sequence from this region rather than identifying an insertion or translocation specific to these ALL cases. Our results significantly extend previous efforts to determine the genomic sequence of Jak3 and analyze its expression in childhood pro-B ALL and other forms of ALL.     

Population pharmacokinetics of methotrexate in Mexican pediatric patients with acute lymphoblastic leukemia

Cancer Chemotherapy and Pharmacology - To develop and validate a population pharmacokinetic model of Methotrexate (MTX) in Mexican children with acute lymphoblastic leukemia (ALL) for the design of...     

High expression of Myosin 1g in pediatric acute lymphoblastic leukemia

Acute Lymphoblastic Leukemia (ALL) is the most frequent cancer in pediatric population. Although the treatment has improved and almost 85% of the children are cured about 20% suffer relapse, therefore finding molecules that participate in the pathogenesis of the disease for the identification of relapse and patients at risk is an urgent unmet need. Class I myosins are molecular motors involved in membrane tension, endocytosis, phagocytosis and cell migration and recently they have been shown important for development and aggressiveness of diverse cancer types, however Myo1g an hematopoietic specific myosin has not been studied in cancer so far. We evaluated the expression of Myo1g by qRT-PCR, Immunocytochemistry and Immunofluorescence in a cohort of 133 ALL patients and correlated the expression at diagnosis and after treatment with clinical features and treatment outcomes. We found high expression levels of Myo1g in Peripheral Blood Mononuclear Cells (PBMCs) from patients with ALL at diagnosis and those levels decreased after complete remission; furthermore, we found an increase in Myo1g expression on patients with 9:22 translocation and those who relapse. This study show that Myo1g is over expressed in ALL and that may participate in the pathogenesis of the disease specially in high-risk patients.