miR-181a在急性淋巴细胞白血病患儿中的表达及其功能研究

目的 检测microRNA-181a(miR-181a)在急性淋巴细胞白血病(ALL)患儿中的表达水平,并研究其在ALL细胞株CCRF-CEM细胞及耐药株CEM-C1细胞中的功能.方法 采用实时荧光定量PCR方法检测ALL患儿骨髓样本、ALL细胞株CCRF-CEM细胞及其耐药株CEM-C1细胞中miR-181a的表达水平.采用电穿孔转染的方法抑制耐药株CEM-C1细胞并上调非耐药株CCRF-CEM细胞中miR-181a的表达,予不同浓度梯度(终浓度分别为0.01、0.1、1、10、100、1 000 ng/ml)喜树碱处理后,采用CCK-8法观察各浓度梯度喜树碱处理后细胞的存活情况,绘制细胞增殖抑制曲线并计算半数抑制浓度(IC50).结果 初诊-复发组患儿初诊及复发骨髓样本miR-181a相对表达水平(4.84±2.71及6.53±2.20)均高于对照组(1.41±0.53)(P=0.017、0.001),初诊-完全缓解组患儿初诊骨髓样本miR-181a水平(7.58±2.50)较对照组明显升高(P=0.000),而完全缓解后miR-181a水平下降至1.35±0.35,与对照组比较差异无统计学意义(P=0.863).CEM-C1细胞miR-181a相对表达水平(-4.39±0.08)较CCRF-CEM细胞(-2.32±0.03)明显升高(P=0.000).转染miR-181a抑制剂CEM-C1细胞较转染阴性对照组增殖抑制率明显升高(P＜0.05),IC50分别为30.61、2 255.00 ng/ml,耐药指数(RI) =73.67.miR-181a过表达CCRF-CEM细胞较阴性对照组增殖抑制率明显降低(P＜0.05),IC50分别为126.60、1.34 ng/ml,RI=94.26.结论 ALL患儿骨髓及CEM-C1细胞中miR-181a异常高表达,抑制CEM-C1细胞中miR-181a的表达可明显增加CEM-C1细胞的药物敏感性,在CCRF-CEM细胞中上凋miR-181a的表达能明显增加CCRF-CEM细胞的耐药性.     

血浆miR-221在急性髓细胞白血病中的表达及意义

目的:探讨血浆miR-221在急性髓细胞白血病(acute myeloid leukemia,AML)中的表达及意义.方法:收集38例成人AML患者,其中初发患者12例(M2 1例,M3 3例,M4 6例,M5 2例),复发患者8例(M2 2例,M3 3例,M4 2例,M5 1例),完全缓解(complete remission,CR)患者18例(M2 2例,M3 6例,M4 6例,M5 4例)及3例正常人的外周血血浆标本.采用基于SYBR Green荧光染料法的茎环实时定量PCR(qRT-PCR),以U6为内参照,用2(-ΔCt)方法计算miR-221的相对表达量,分析血浆miR-221的表达情况.结果:qRT-PCR结果显示,较之CR组和正常对照组,AML初发组和复发组中血浆miR-221的表达量明显升高,差异均有显著性(P＜0.05).初发组和复发组之间无统计学差异(P＞0.05).CR组比正常对照组的表达量高,但两组差异并无统计学意义(P＞0.05).结论:血浆miR-221在AML初发和复发患者中的表达量高,在CR患者及正常对照中的表达量低,提示血浆miR-221与AML肿瘤负荷呈正相关,微创的血浆miRNAs检测可作为临床微小残留病检测的新方法.     

Advances in immunotherapy for pediatric acute myeloid leukemia

Introduction: Achieving better disease control in patients diagnosed with acute myeloid leukemia (AML) has proven challenging. Overall survival has been impacted by addressing treatment related mortality with focused supportive care measures. Despite this improvement, it remains difficult to induce durable leukemia remissions despite aggressive chemotherapeutic regimens. The addition of hematopoietic stem cell transplants (HSCT) has allowed further treatment intensification and provided the benefit of graft-versus-leukemia (GVL) effect. However, HSCT carries the risk of transplant related morbidities, particularly GVHD, and anti-tumor responsiveness is still suboptimal. Thus, there is a need for alternate therapies. Immunotherapy has the potential to address this need.

Areas covered: This review highlights promises and challenges to immune-based therapies for AML. It aims to summarize immunotherapeutic strategies trialed in AML patients to date, inclusive of: antibodies, vaccines, and cellular therapy. It emphasizes those being used in the pediatric population, but also includes adult clinical trials and translational science that may ultimately extend to pediatric patients.

Expert opinion: The elusiveness of an ideal surface antigen target together with an immunosuppressive leukemic microenvironment add to the already difficult challenge in developing AML-targeted immunotherapies. Though many hurdles remain, recent translational discovery and progressive clinical advances anticipate exciting future developments.     

DNMT3A基因对非M3型急性髓系白血病患者预后的预测价值

目的通过分析非M3型急性髓系白血病(AML)患者的临床和实验室资料,进一步阐明DNMT3A基因突变在非M3型AML患者预后中的意义及其影响AML发生发展的可能机制。方法采用R语言3.5.1版本RTCGAToolbox包下载癌症基因组图谱数据库180例非M3型AML患者临床及突变信息,数据为开放性数据。将患者按照突变状态进行分组,比较各组患者外周血白细胞计数、骨髓原始细胞比例、无事件生存期以及总体生存期有无差别。将患者按照年龄分为<60岁组和≥60岁组,使用Kaplan-Meier方法绘制生存曲线,并运用Log-rank检验进行<60岁和≥60岁患者、DNMT3A突变组及野生组患者生存率的比较。采用Cox比例风险模型进行DNMT3A突变和其伴随突变及年龄分层等因素对患者总体生存期的单因素和多因素分析。结果共纳入非M3型AML患者180例,DNMT3A突变组与野生组患者临床特征比较发现突变组白细胞计数显著高于野生组(Z=-2.606,P=0.009),且DNMT3A突变多见于中危患者。值得注意的是,DNMT3A突变常伴随FLT3(P=0.025)、NPM1(P<0.001)、IDH1(P=0.002)突变的发生,且DNMT3Awt/FLT3wt组无事件生存期显著高于DNMT3Amut/FLT3mut组(11.00个月vs 6.15个月,P=0.005),而与DNMT3Amut/FLT3wt、DNMT3Awt/FLT3mut组之间的差异无统计学意义。Cox多因素分析结果显示,高龄是AML患者总体生存的独立危险因素(HR=2.974,95%CI:1.966~4.500,P<0.001);另外,DNMT3A R882突变是AML患者预后不良的独立预测因子(HR=1.937,95%CI:1.179~3.182,P=0.009)。结论高龄(≥60岁)和DNMT3A R882突变为预后不良的独立预测因子。DNMT3Amut/FLT3mut亚型与DNMT3Awt/FLT3wt、DNMT3Amut/FLT3wt、DNMT3Awt/FLT3mut相比,临床预后更差。     

FANCD1在急性髓系白血病细胞中的表达

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急性髓系白血病患者131例免疫表型与预后的相关性分析

目的 探讨急性髓系白血病(AML)免疫表型与预后的相关性.方法 采用多色流式细胞术对131例AML患者进行检测,分析其免疫表型与患者年龄、初诊时WBC、PLT和Hb的关系,及其对完全缓解率(CR)的影响.结果 AML患者中髓系抗原表达阳性率最高的是CD13、CD33和髓过氧化物酶(MPO),急性早幼粒细胞白血病(M3)亚型中CD34和HLA-DR表达率较低,淋巴细胞抗原CD19和CD7表达最常见,阳性率分别为15.4%和14.6%.CD7阳性组患者的年龄明显高于阴性组患者年龄(t=-2.27,P＜0.05),CD14阳性组患者的初诊WBC计数明显高于阴性组(Z=-2.284,P＜0.05).131例AML患者的总CR率为56.5%,CD34阳性组(82例)的CR率为45.1%,CD34阴性组(49例)的CR率为75.6%,两组间差异有统计学意义(x2=11.524,P＜0.05).CD34和HLA-DR双阳性组(74例)的CR率为41.9%,单阳性组(38例)CR率为78.9%,双阴性组(19例)CR率为68.4%,3组问差异有统计学意义(Z=-3.492,P＜0.01).CD7、CD19、CD13、CD33、CD38、CD15、CD64、CD14及MPO等抗原表达阳性组和阴性组间CR率的差异无统计学意义(P均＞0.05).多因素回归分析显示,患者年龄大于60岁、初诊时WBC大于50×109/L、PLT大于30×109/L、Hb小于60 g/L以及CD34阳性是低CR率的独立风险因素.结论 AML患者年龄,初诊时WBC、PLT和Hb计数以及CD34表达与CR率有关,免疫表型的检测对于判断AML预后有一定价值,有助于指导临床治疗和判断预后.     

急性髓性白血病全基因组miRNA表达谱研究

目的通过比较微小RNA(miRNA)在急性髓性白血病(AML)与对照组骨髓样本中的表达情况,发现在AML中异常表达的miRNA。方法提取骨髓样本总RNA,应用Illumina高通量测序技术对15例AML患者(AML组)和10例骨髓象正常的贫血或发热原因待查患者(对照组)骨髓样本中的miRNA表达水平进行检测,分析比较二者miRNA表达的差异。结果AML样本与对照样本间差异表达的miRNA共307种,其中232种miRNA在AML组呈表达上调,75种miRNA呈表达下调。结论多种miRNA在AML患者的骨髓样本中异常表达,提示它们可能在AML发生和发展过程中起重要调控作用。     

Dysregulation of miR-124-1 predicts favorable prognosis in acute myeloid leukemia

ObjectiveMicroRNA miR-124 has been suggested as a tumor suppressor for its role in inhibiting cell growth, inducing differentiation and promoting apoptosis. The present study was aimed to investigate the expression status of miR-124-1 and its clinical relevance in patients with acute myeloid leukemia (AML).Designs and methodsReal-time quantitative PCR was performed to detect the expression level of miR-124-1 in AML patients. The clinical significance of miR-124-1 expression in AML was investigated.ResultsmiR-124-1 underexpression was identified in 30 (36%) of 83 AML patients. No significant difference could be observed in sex, age and blood parameters between the patients with and without miR-124-1 underexpression. The frequency of miR-124-1 underexpression was higher in the patients with t(15;17) than in others (62% versus 30%, P = 0.040). The status of miR-124-1 expression was not correlated with the mutations of nine genes (FLT3-ITD, NPM1, C-KIT, IDH1/IDH2, DNMT3A, N/K-RAS and C/EBPA). The patients with miR-124-1 underexpression had borderline longer overall survival and relapse-free survival than those without miR-124-1 underexpression (P = 0.052 and 0.045, respectively).ConclusionsThese findings suggest that miR-124-1 underexpression is a common event and might have a favorable impact on prognosis in AML.     

Long noncoding RNA ZFAS1 enhances adriamycin resistance in pediatric acute myeloid leukemia through the miR-195/Myb axis

Background: Development of chemoresistance remains a major obstacle for pediatric acute myeloid leukemia (AML) management. Zinc finger antisense 1 (ZFAS1) is a novel tumor-related lncRNA that has been reported as an oncogene involved in the development of pediatric AML. The purpose of the present study was to investigate the role and underlying mechanism of ZFAS1 in AML chemoresistance. Methods: The expression levels of ZFAS1 and miR-195 were assessed by qRT-PCR and Myb expression was detected using western blotting. The CCK-8 assay was used to determine the IC₅₀ value for adriamycin (ADR) and cell proliferation. Cell apoptosis was measured by flow cytometry. The targeted interaction between miR-195 and ZFAS1 or Myb was evaluated by the dual-luciferase reporter assay or RNA immunoprecipitation assay. Results: Our data revealed that ADR treatment induced ZFAS1 expression in pediatric AML. Silencing of ZFAS1 or Myb alleviated AML cell resistance to ADR in vitro. ZFAS1 directly targeted miR-195 and negatively modulated miR-195 expression. Myb was a direct target of miR-195. Moreover, the inhibitory effect of ZFAS1 silencing on ADR resistance of AML cells was mediated by miR-195 in vitro. Myb was involved in the regulation of the ZFAS1/miR-195 axis in ADR resistance of AML cells. Conclusion: Our data indicated that ZFAS1 silencing alleviated ADR resistance of AML cells in vitro through acting as a sponge for miR-195 and regulating Myb expression. Targeting ZFAS1 might be a promising therapeutic strategy for pediatric AML treatment.

Long noncoding RNA ZFAS1 silencing alleviated ADR resistance of AML cells.     

高超二倍体、三/四倍体染色体核型在急性髓细胞白血病中的预后意义

目的描述高超二倍体、三/四倍体染色体核型急性髓细胞白血病(HH/TT-AML)患者的生物学特征,探讨其与预后的关系。方法回顾性分析2006年3月至2017年6月28例HH/TT-AML初治患者一般临床特征、治疗情况,用Kaplan-Meier方法分析预后影响因素。结果 HH/TT-AML患者核型以49条染色体为主,占39.3%(11/28),50～55条染色体次之,占32.1%(9/28)。高超二倍体核型急性髓细胞白血病(HH-AML)患者以+8(77.3%,17/22)、+21(54.5%,12/22)多见。生存分析显示,伴-5/5q-、-7/7q-、-17/der(17p)及der(3q)的HH/TT-AML患者总体生存率劣于不伴此类异常者(4.1月vs 10.1月,P0.05);三/四倍体染色体核型急性髓细胞白血病(TT-AML)患者总体生存率与HH-AML患者差异无统计学意义(8.4月vs 7.2月,P0.05);异基因造血干细胞移植患者总体生存率优于仅化疗患者(25.4月vs 4.1月,P0.01)。结论 HH/TT-AML患者存在较大异质性,不良预后相关染色体异常的识别有助于预后分层。此类患者总体生存期短,联合化疗获缓解后尽早行异基因造血干细胞移植能显著改善患者预后。     

Beginning treatment for pediatric acute myeloid leukemia: the family connection

There is a loud silence on psycho-oncology research in relation to pediatric Acute Myeloid Leukemia (AML). This article is part of a series that begins to address the psycho-social hiatus. The present article documents the less obvious, often hidden, aspect of beginning treatment for pediatric AML--the "behind the scenes" experience of the home and family connection. The findings are from the first stage of a five year longitudinal study that examines through qualitative research the experience of childhood leukemia from the perspective of the child, siblings and parents. Open-ended interviews, audio-recorded and transcribed verbatim, were thematically analyzed with the assistance of the Non-numerical Unstructured Data by processes of Indexing Searching and Theory-building (NUD*IST) computer program. The findings emphasize the disruption to normalcy in relation to home life, school, and work, which is exacerbated for families who relocate for specialist treatment. The findings emphasise the need for support for families coping with childhood AML.     

急性髓系白血病预后相关基因表达谱

目的运用基因芯片技术,研究首次完全缓解持续时间(CCR1)相差显著的急性髓系白血病(AML)M2a亚型之间基因表达谱差异,寻找影响AML-M2a预后相关基因。方法应用Agilent Human 1B寡核苷酸基因芯片,检测3例CCR1 6个月内复发(A组)和3例CCR1 12个月以上(B组)的AML-M2a患者初诊时骨髓单个细胞基因表达谱及其差异。结果在检测的20173个基因中,筛选出共同差异表达基因共22个,其中在A组各例都表达上调的基因有10个,同时表达下调的基因有12个。结论APP等22个基因的表达可能与接受常规方案标准剂量化疗的AML-M2a首次完全缓解持续时间有关,可能成为早期诊断难治性AML的指标。     

Response in acute myeloid leukemia

For many years, response to induction therapy in acute myeloid leukemia was classified as "complete response" (CR) or "no CR." The emphasis on CR reflected the proven ability of CR to extend survival, the outcome of primary interest to patients. Beginning with the adoption of complete response with incomplete platelet recovery (CRp), recent years have seen the acceptance of responses less than CR as beneficial. Although these responses denote that a drug is active, relatively little attention has been paid to the effect of such responses on survival. This article explores the effect of such responses on survival.     

儿童B-急性淋巴细胞白血病治疗中微量残留病动态监测及其意义

目的评价微量残留病(MRD)监测在儿童B-急性淋巴细胞白血病(B-ALL)治疗中的预后价值。方法2001年9月1日至2004年10月31日采用ALL-XH-99方案行MRD标记筛选并监测的B-ALL患儿102例。用四色多参数流式细胞仪监测诱导治疗结束完全缓解(CR)时、髓外白血病预防性治疗前、早期强化前、维持治疗中定期强化前、维持治疗1年及2年时患儿。MRD 。结果①行筛选并监测的102例B-ALL患儿中,对诱导治疗结束获CR的86例患儿进行了MRD监测。其中MRD ≥10~(-4)的患儿20例,占23．30％;MRD<10~(-4)的患儿66例,占76．70％,其39个月无事件生存率分别为 0．00％和(83．00±9．90)％,差异有统计学意义(P<0．01)。②单因素分析显示患儿的性别、起病时年龄、白细胞计数及染色体倍体数与CR时MRD水平无关(P>0．05);而Ph染色体、诱导治疗第19天骨髓幼稚淋巴细胞比例、CR时间、ALL-XH-99危险度分类标准与其相关(P<0．05)。③多因素分析显示 CR时MRD水平具有独立的预后价值(比例风险为5．381,95％可信区间为0．004-0．624,P<0．05)。结论诱导治疗结束CR时MRD水平可用于评估早期治疗反应,是儿童B-ALL治疗中重要的预后因素之一。     

Adult acute myeloid leukemia

Acute myeloid leukemia (AML) is a group of several different diseases, the treatment and outcome of which depend on several factors, including leukemia karyotype, patient age, and comorbid conditions. Despite advances in understanding the molecular biology of AML, its treatment remains challenging. Standard regimens using cytarabine and anthracyclines for induction followed by some form of postremission therapy produce response rates of 60% to 70%, with less than 20% of all patients achieving long-term disease-free survival. New therapies are emerging based on the definition of specific cytogenetic-molecular abnormalities. Such targeted therapies offer the promise of better antileukemic activity in adult AML.     

三例急性髓系白血病患者基因表达谱的演变

目的研究急性髓系白血病（AML）患者初诊与复发时的基因表达谱差异，探讨难治性AML的发病机制。方法应用Agilent Human 1B寡核苷酸基因芯片，动态检测了3例AML-M2a患者初诊、复发时骨髓单个核细胞的基因表达谱差异。结果在检测的20173个基因中，有10个基因在3例患者初诊、复发时共同差异表达，其中7个基冈在复发时均共同上调，3个基因在复发时均表现下调。结论DAPKI等10个基因的表达变化可能与AML-M2a发病和复发有关，这些新基因的发现可能对早期诊断难治性AML具有重要价值，同时为难治性AML提供新的治疗靶点。