Expression of BCL2L12and LIPOCALIN 2 in Adult Patient with Acute Myeloid Leukemia and Correlation with Clinical Response

Background: Acute myeloid leukemia (AML) is a malignancy arising within the bone marrow (BM), in which leukemic cells proliferate uncontrollably in association with a disruption of normal hematopoiesis.Aim of the work to evaluate the expression of LCN and BCL2L2, in newly diagnosed bone marrow samples from adult with AML and to correlate their expression levels with clinical and Laboratory data of the patients especially that known to have a prognostic feature. Methods: This study was carried out on 87 consecutive newly diagnosed adult AML patients of which 75 are evaluated for both LCN, BCL2L12 (All 87 are evaluated for LCN). In addition, 20 donors of matched age and sex healthy individuals from donors for bone marrow transplantation were included as a control group. Results: No statistical significant correlation was found between LCN over-expression and the control group , there was no statistical significance between its expression and age, sex, hepatomegally, splenomegally and lymphadenopathy, also there was no statistical significance regarding peripheral blood and bone marrow findings, immunophenotyping, cytogenetics or molecular findings and MRD at day 15.No statistical significance was found between BCL2L12 expression and the control group again there was no statistical significance between its expression and age, sex, hepatomegally, splenomegally and lymphadenopathy, also there was no statistical significance regarding peripheral blood and bone marrow findings, immunophenotyping, cytogenetics or molecular findings and MRD at day 15. Conclusion: LCN and BCL2L12 were found to be expressed with non significant difference in AML as in normal subjects; however studies on large number of cases are needed to confirm our finding. The role of LCN and BCL2L12 need to be verified by further large-scale sample and further studies.     

Retreatment of Chronic Lymphocvtic Leukemia with 2-Chlorodeoxyadenbsine (CdA) at Relapse Following CdA-induced Remission: No Acquired Resistance

B-cell chronic lymphocytic leukemia (CLL) initially responds well to treatment with alkyl-ating agents, but subsequently resistance may develop. We recently showed that the nucleo-side analogue 2-chlorodeoxyadenosine (CdA) produce a high remission rate in previously treated patients with symptomatic CLL. We report here the results of retreatment with CdA given to 6 previously CdA-treated patients who had progression off therapy. Two initially had a complete remission and 4 a partial remission. The median time from the start of initial CdA-treatment to retreatment was 19 months (range 8-28 mos). The tumor cell response to CdA was assessed by calculating the elimination rate of circulating leukemia cells. The lym phocyte half-life varied in the different patients, but was similar in all six patients when comparing the first CdA-course with CdA-treatment at relapse, with a correlation coefficient of r² = 0.89, However, the toxicity towards thrombopoiesis was greater at relapse than during the initial treatment in four of the six patients. CLL cells do not seem to acquire resistance to CdA, but retreatment with CdA at relapse following CdA-induced remission is hampered by more severe drug-related cytopenia. Thus, the quality of subsequent remission may be reduced, despite continuing sensitivity of the leukemia cells to CdA.     

急性白血病Bcl-2、P170、CD_(34)表达及临床预测价值的研究

目的分析和评估急性白血病Bcl－2、P170、CD_(34)表达的临床预测价值。方法用ABC免疫细胞化学法检测42例急性白血病Bcl－2、P170、CD_(34)的表达。结果Bcl－2、P170、CD_(34)高表达的急性白血病完全缓解（CR）率低于低表达或不表达者（45．5％V77．4％,50．0％v78．6％,57．1％v75．0％,P＜0．05）,未CR和早期复发率高,分别为81．8％,78．6％,64．3％。化疗2个疗程达CR者Bcl－2、P170的高表达率高于化疗1个疗程达CR者（36．4％v5．6％,45．5％v11．1％,P＜0．05）。Bcl－2与急性白血病预后的阳性符合率为90．9％,略高于P170、CD_(34)。同时检测3项指标的阳性、阴性及总符合率分别为100％、80％及84．6％。结论Bcl－2、P170、CD_(34)的高表达均预示急性白血病的预后不良。Bcl－2可作为预测急性白血病疗效、预后较敏感的指标,多指标联合检测预测价值更高。     

非霍奇金淋巴瘤侵犯骨髓及合并淋巴瘤细胞白血病的临床分析

目的:了解非霍奇金淋巴瘤骨髓侵犯的特征与病理类型、临床表现、治疗及预后的关系。方法:413 例经病理学诊断为非霍奇金淋巴瘤患者经骨髓穿刺检查后,对112例诊断为骨髓侵犯患者的骨髓病理形态学特征、骨髓侵犯程度与临床表现、分期,疗效以及生存时间的关系进行分析。结果:骨髓侵犯的发生率为27．12％,骨髓侵犯的形态学特征与原发肿瘤的一致;骨髓侵犯以B细胞淋巴瘤多见,多发生在晚期;骨髓侵犯程度与临床表现、疗效、预后明显相关,骨髓侵犯越重,临床症状越重,预后不良。结论:非霍奇金淋巴瘤骨髓侵犯的形态学特征与原发肿瘤细胞的形态一致,骨髓侵犯程度越重,临床出现各种相关症状重,化疗的缓解率低,疗效差,生存期明显缩短,预后不良,但仍有治愈的机会。     

突变型p53、bcl-2在急性白血病中的表达及与细胞周期关系的研究

目的：研究急性白血病(AL)中p53、bcl-2的表达特性及相关性，分析其表达与细胞周期的关系。方法：49例AL患者为实验组，同期住院的骨髓象完全正常的非血液系恶性疾病患者14例为对照组，取骨髓标本，分离单个核细胞，用特异性单克隆抗体标记。流式细胞仪检测突变型p53及bcl-2基因蛋白表达，其中有26例患者同时用流式细胞仪测定了细胞周期。结果：AL组p53、bcl-2蛋白表达阳性率分别为44.9％、49.0％，显著高于对照组，ALL组与ANLL组差异无显著性，M3组显著低于其他非M3 ANLL组。突变型p53蛋白与bcl-2蛋白表达呈正相关，二者在白血病的发病上有协同作用。白血病组比对照组细胞周期中S+G2／M期细胞数低，p53、bcl-2表达与细胞周期无关。结论：突变型p53、bcl-2有致白血病的作用，二者在白血病的发展中可能起一定协同作用。联合检测突变型p53及bcl-2对预测白血病的预后有一定价值。     

Correlation of Inhibin and Several Antioxidants in Children with Acute Lymphoblastic Leukemia

Background: Acute lymphoblastic leukemia (ALL) is most common in childhood. Inhibin (a non-steroidal glycoprotein hormone of gonadal origin) can be used as marker of fertility. The current study was conducted to evaluate inhibin levels in ALL patients and to estimate its correlation with some antioxidants in these in comparison with control subjects. Materials and Methods: This study was conducted on sixty patients with ALL and thirty children as controls. Fasting blood samples were taken from each subject and analyzed for haemoglobin, serum protein, vitamin E and C, in addition to glutathione and inhibin. Results: The results of the study showed highly significant decreases (p<0.001) in haemoglobin, glutathione and inhibin levels with significant decreases (p<0.05) in serum protein and vitamin E levels for patients group in comparison with controls while there was no significant differences in vitamin C. Moreover, there were significant correlations between inhibin levels and serum protein, glutathione and both vitamins (E and C) in the ALL patient group (r= 0.81, 0.80, 0.77 and 0.69, respectively). Conclusions: The present results indicated infertility in patients with ALL demonstrated by low inhibin level as a consequence of abnormality in anti-oxidative metabolism due to the cancer process. So, it can be suggested the need for routine measurement of inhibin for leukemic patients to estimate the action of hormones of gonadal origin.     

Clinical and Molecular Characteristics of Patients with Mixed Phenotype Acute Leukemia

Introduction: Mixed phenotype acute leukemia (MPAL) is a rare heterogeneous disease with a poor prognosis. This study analyzed the clinical, immunophenotypic, molecular, and cytogenetic characteristics of a group of patients with MPAL. Methods: This prospective study included 75 patients diagnosed with MPAL according to the World Health Organization (WHO)-2016 diagnostic criteria, using cytochemistry, conventional cytogenetics, and molecular studies. Screening of BCR::ABL1 fusion gene was performed by Fluorescent in-situ hybridization (FISH) and polymerase chain reaction (PCR). Results: Children represented 49.3% of MPAL patients. The main phenotype was B-lymphoid/myeloid (80%). Molecular alterations were detected in 17 patients (22.7%). The BCR::ABL1 fusion gene was detected in 10 patients (13.3%).. Myeloid protocols were used to treat 58 patients (77.3%), and lymphatic protocols in 17. By the end of the follow-up, 57 patients (76%) achieved complete remission (CR). There was no association between BCR::ABL1 and response to treatment. The cumulative overall survival (OS) at 12 months was 47.8%. The bone marrow transplantation (BMT) was associated with better OS (p = 0.027). The disease-free survival (DFS) was not affected by all tested prognostic factors. Conclusion: MPAL is a complex entity with heterogeneous features. BCR::ABL1 is a common abnormality. BMT is associated with better OS.     

Interleukin 2 and the Treatment of Leukemia and Lymphoma

As evidence has accumulated that allogeneic bone marrow transplantation provides therapeutic benefit by means of a graft versus malignancy effect, there has been a corresponding increase in interest in inducing or enhancing such an effect after chemotherapy and/or autologous bone marrow transplantation. Administration of Interleukin-2 may be one way of achieving this aim. Recent studies have shown the cytokine is tolerated after ABMT/chemotherapy in immunomodulatory doses and that the MHC unrestricted cytotoxic effector mechanisms induced can indeed discriminate between normal and malignant tissue. As always, larger scale randomized studies will be required before the therapeutic efficacy of this approach can been assessed.     

Evaluation of BAX and BCL-2 Gene Expression and Apoptosis Induction in Acute Lymphoblastic Leukemia Cell Line CCRFCEM after High- Dose Prednisolone Treatment

Objective: Glucocorticoids are one of the most important drugs in the treatment of acute lymphoblastic leukemiafor children. It is very important to response to glucocorticoid in the prognosis of these patients. Therefore, resistanceto treatment is a major problem in lymphoid leukemia cases. In, this study, CCRF-CEM cell line was selected as achemotherapy-resistant model. The aim of this study was to evaluate the effect of high dose prednisolone on inductionof apoptosis and changes in BAX and BCL-2 gene expression at different times. Methods: CCRF-CEM cell lines weregrown in standard conditions. Based on previous studies, a dose of 700 μM as subtoxic dose was selected. Changes ingene expression of BAX and BCL-2 were evaluated by using real time PCR techniques. Also stained with annexin Vand the induction of apoptosis was assessed by FACS machine. Results: In this study it was found that prednisolone inhigh doses at different times significantly increased the gene expression of BAX and on the other hand the amount ofBCL-2 expression was reduced. Cells that treated for 48 hours had more significant changes in gene expression. Basedon flowcytometry data, prednisolone can induce apoptosis in a time dependent manner on this cancerous resistant cellline. Conclusions: Apoptosis induced by high-dose prednisolone in the CCRF-CEM cells, which is almost resistant,and possibly mediated by reducing the expression of BCL-2 and BAX up-regulation.     

Increased Risk of Thai Childhood Acute Lymphoblastic Leukemia with the MiR196a2 T>C Polymorphism

Objectives: This study assessed associations of the miR196a2 (rs11614913) T>C polymorphism withsusceptibility to childhood acute lymphoblastic leukemia (ALL) and clinical outcomes. Materials and Methods: Blood DNA samples from 104 childhood ALL patients and 180 healthy children were studied for the miR-196a2 (rs11614913) polymorphism using a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) approach. Results: The frequency of the miR-196a2 (rs11614913) T allele in controls was 0.51 compared with 0.33 in ALL cases. In this study, CC, TC heterozygote and CC/TC genotypes were significantly associated with increase childhood ALL susceptibility compared with the TT wild type (OR =4.321, 95% CI = 2.091-8.930 p=0.000, OR = 2.248, 95% CI =1.103-4.579, p=0.024, OR = 2.921, 95% CI = 1.504-5.673 p=0.001, respectively). However, the miR-196a2 (rs11614913) T>C polymorphism was not associated with demographic data or clinico-pathological data in ALL cases. Conclusion: CC, TC and CC+TC genotypes of miR-196a2 (rs11614913) was significantly associated with increased susceptibility in Thai childhood ALL but not with clinical variables.     

Association between the MDM2 T309G Polymorphism and Leukemia Risk: a Meta-analysis

Several studies have suggested associations between MDM2 (mouse double minute 2 homolog) polymorphisms and leukemia risk, but they reported contradictory results. For better understanding of the effect of MDM2 T309G polymorphism on leukemia risk, we performed a meta-analysis. All eligible studies were identified through a search of PubMed, Web of Science, EMBASE, and Chinese Biomedical Literature (CBM) databases before May 2014. Assessment of associations between the MDM2 T309G polymorphism and leukemia risk was conducted by odds ratios (ORs) and 95% confidence intervals (95% CIs). Finally, a total of 11 publications covering 12 case-control studies with 2, 362 cases and 5, 562 controls concerning MDM2 T309G polymorphism with respect to leukemia were included in the meta-analysis. Significant associations were found between MDM2 T309G polymorphism and leukemia risk in four models in overall populations (G vs T: OR=1.29, 95% CI=1.11-1.49, p=0.001; GG vs TT: OR=1.67, 95% CI=1.21-2.30, p=0.002; GG vs TG/TT: OR=1.56, 95% CI=1.21-2.00,p=0.001; GG/TG vs TT: OR=1.28, 95% CI=1.05-1.57, p=0.015). In the sub-group analysis according to ethnicity, increased leukemia risks were observed in three genetic models among Asians but not Caucasians. In conclusion, the results of our meta-analysis suggest that the MDM2 T309G polymorphism can increase the risk of leukemia, especially among Asian populations.     

Clinical pharmacology of cytarabine in patients with acute myeloid leukemia: a Cancer and Leukemia Group B study

The pharmacokinetics of cytarabine (ara-C) were determined in 265 patients with acute myeloid leukemia (AML) receiving ara-C (200 mg/m² per day for 7 days as a continuous infusion) and daunorubicin during induction therapy. The mean (standard deviation) ara-C concentration at steady-state (Css) and systemic clearance (Cl) were 0.30 (0.13) M and 134 (71) l/h per m² respectively. Males had a significantly faster ara-C Cl (139 vs 131 l/h per m²,P=0.025) than females. Significant correlations were noted between ara-C Cl and the pretreatment, peripheral white blood cell count (P=0.005) and pretreatment blast count (P=0.020). No significant differences in ara-C Css or Cl were noted in patients achieving complete remission compared with those failing therapy (P=0.315,P=0.344, respectively). No significant correlations were observed between ara-C pharmacokinetic parameters and several indices of patient toxicity. Our findings indicate that variability in ara-C disposition in plasma at this dosage level does not correlate with remission status or toxicity in patients with AML receiving initial induction therapy with ara-C and daunorubicin.This study was supported in part by grants from the National Cancer Institute (CA-03927, CA-37027, CA-59518, CA-47577, CA-32291, CA-07968, CA-16450, CA-16450, CA-26806, CA-47545, CA-12197, and CA-41287), the Upjohn Company, and the Coleman Leukemia Research Fund     

白血病患者P2X7受体的功能性表达分析

目的研究P2X7受体在白血病患者中的功能性表达及与白血病治疗的关系。方法采用半定量RT-PCR、流式细胞术和细胞内钙离子测定的方法,从核酸、蛋白和功能水平研究白血病患者骨髓单个核细胞（BMMC）P2X7受体的表达和功能。结果白血病患者和正常人标本中可检出P2X7受体的表达,核酸、蛋白和功能水平结果一致。AML、ALL、CML患者P2X7受体表达阳性率高于正常人;但AML各亚型中P2X7受体阳性强度存在差异,M2组低于正常人,M5,M6组高于正常人;按照P2X7受体阳性强度分组,P2X7受体高表达患者诱导治疗后的完全缓解率显著低于P2X7受体阴性和P2X7受体低表达患者。结论P2X7受体在白血病患者BMMC中有功能性表达,其高表达与患者治疗不敏感相关。     

急性白血病血浆内皮素水平及其对纤溶系统的影响

为研究急性白血病患者血浆内皮素水平及其对纤溶系统的影响。方法测定了５３例急性白血病性白血病患者ＥＴ及纤溶指标ＰＡ、ｔＰＡ：Ａ、ＰＡＩ：Ａ、Ｄ－Ｄ。结果急性白血病患者ＥＴ水平明显高于正常,以初治及未缓解组ＥＴ升高显示,完全缓解组较初治及未缓解组明显降低。     

Heterogeneity of B Lineage Acute Lymphoblastic Leukemias (B-ALL) with Regard to their in Vitro Spontaneous Proliferation, Growth Factor Response and BCL-2 Expression

The spontaneous proliferation and the effects of 8 various growth factors (GF) were evaluated on leukemic cells from 27 patients with B-lineage ALL. Two groups of ALLs were distinguished: ALLs from group I (21 patients) exhibited a low spontaneous proliferative rate and were stimulated by IL-3 + IL-7 SCF and/or LIF, while ALLs from group II (6 patients) had a high spontaneous proliferative rate and did no longer require this combination of GFs for proliferation. No effect of bFGF, IGF-I, IL-10 and IL-11 alone or in combination, was observed. Such differences in the behaviour of B-ALLs indicated that the GF requirement of ALL blasts was not related to the presence of serum in the culture nor to the pattern of reactivity of ALL blasts for B lymphoid markers or CD34 antigen. Furthermore, we showed in 1/9 cases that high proliferation might be due to an overexpression of the bcl-2 proto-oncogene and to the acquisition of an autocrine secretion.     

Tyrosine Phosphorylation of the Proto-oncogene Product Vav and Its Association with the Adapter Grb2/Ash in a Human Leukemia Cell Line UT-7

The vav proto-oncogene product (Vav) is expressed exclusively in hematopoietic cells and is reported to have guanine nucleotide exchange activity. Here we report that granulocyte-macrophage colonystimulating factor, interleukin-3, and erythropoietin induce tyrosine phosphorylation of Vav in a human leukemia cell line UT-7. Tyrosine phosphorylation of Vav is rapid and transient; it occurs within 1 min of the stimulation and at physiological concentrations of the factors. Furthermore, we show that Vav is constitutively associated with the adapter molecule Grb2/Ash in UT-7. These data suggest that tryosine kinases, the adapter Grb2/Ash, and the guanine nucleotide exchange factor Vav are members of a signaling pathway leading to Ras activation in hematopoietic cells.     

食管癌组织中Skp2的表达及其与临床病理特征的关系

目的研究Skp2（S相激酶相关蛋白2）在食管癌组织和正常组织中的表达差异,并探讨Skp2的表达和食管癌临床病理特征的关系。方法选取64例食管癌组织和22例食管良性疾病组织,应用免疫组化S-P法检测Skp2在上述组织中的表达差异,然后用SPSS10．0软件对上述数据进行分析。结果64例食管癌组织中,Skp2的表达阳性率为32．8％,22例食管良性疾病组织中,Skp2的阳性表达率为9．1％,经χ^2检验,该差异具有统计学意义（P〈0．05）。Skp2的表达和食管癌的分化程度存在相关性（P〈0．05）,但和患者的年龄、性别、病灶位置、淋巴结转移和临床分期等无显著相关性（P〉0．05）。结论Skp2在食管癌的发生过程中具有重要作用,并有望作为治疗的新靶点。     

Molecular Response to Imatinib and Its Correlation with mRNA Expression Levels of Imatinib Influx Transporter (OCT1) in Indian Chronic Myeloid Leukemia Patients

Background and objectives: Imatinib mesylate is approved for the treatment of Chronic Myeloid Leukemia (CML). About 20% of patients with CML do not respond to treatment with Imatinib either initially or because of acquired resistance. In addition to mutated BCR-ABL1 kinase, the organic cation transporter1 (OCT1, encoded by SLC22A1) has been considered to contribute to Imatinib resistance in patients with chronic myeloid leukemia (CML). OCT1 has been reported to be the main influx transporter involved in Imatinib uptake into CML cells. To date, only a few studies have been reported on involvement of influx transporters in development of Imatinib resistance. Therefore this study was aimed to determine the expression level of Imatinib uptake transporter (OCT1) in CML patients and to correlate this level with molecular response. Methods: One hundred fifty eight patients on Imatinib were considered for gene expression analysis study for OCT1 gene. Total RNA was extracted from peripheral blood mononuclear cells. Complementary DNAs (cDNAs) were synthesized and Real Time Polymerase Chain Reaction (RQ-PCR) was performed. Results: High OCT1 expression was present in 81 (51.8%) patients and low OCT1 expression was in 77 (48.7%) patients. Low Sokal risk score group have a significantly high OCT1 expression (p=0.048). The rate of molecular response was higher in those with high OCT1 expression than in those with low OCT1 expression (p=0.05). Both event-free survival and median overall survival were significantly shorter in patients with low OCT1 expressions when compared to the patients with high OCT1 expression (p=0.03 and p=0.05). Conclusions: Our findings demonstrated that the mRNA expression level of OCT1 was significantly correlated with molecular response in CML patients. Based on these findings, present study believes that the pre-therapeutic higher expression of OCT1 may help to predict response to imatinib therapy in CML patients.