Hsa-miR-204反义寡核苷酸对急性淋巴细胞白血病细胞的增殖抑制作用

目的:观察抑制hsa-miR-204表达对人急性淋巴细胞白血病细胞（MOLT-4）生长与凋亡的影响。方法:设计合成hsa-miR-204的反义核苷酸序列（AMO-miR-204）并用脂质体转染法将其转染MOLT-4细胞。Q-PCR检测hsa-miR-204的表达量。噻唑蓝（MTT）试验和平板克隆形成实验检测细胞增殖能力;流式细胞术检测细胞早期凋亡率。Transwell检测细胞侵袭能力。结果:AMO-miR-204可抑制hsa-miR-204的表达,以反义核酸浓度为0.6μmol/L时,下调最为明显（P<0.05）。MTT实验示AMO-miR-204的最佳转染抑制浓度为0.6μmol/L。AMO6组细胞平板克隆形成能力明显减弱［（29±8）%］,差异有统计学意义（P<0.05）。流式细胞术检测AMO6组细胞凋亡达(7.47%）。Transwell细胞侵袭能力实验示AMO6组细胞侵袭能力较其余两组减弱。结论:AMO-miR-204能有效抑制MOLT-4细胞的增殖,并促进其凋亡。hsa-miR-204有可能作为急性淋巴细胞白血病（acute lymphoblastic leukemia,ALL）基因治疗的靶基因,同时也为深入揭示ALL的发生和发展机制提供了实验依据。     

Arsenic trioxide and low-dose cytarabine in older patients with untreated acute myeloid leukemia, excluding acute promyelocytic leukemia

BACKGROUND: Acute myeloid leukemia (AML) carries a dismal prognosis in older patients. In this study, the authors evaluated the safety and efficacy of arsenic trioxide combined with low-dose cytarabine in untreated patients aged >or=60 years with AML. METHODS: In a phase 1/2 design, arsenic trioxide was administered intravenously at a dose of 0.25 mg/kg on Days 1 through 5 and on Days 8 through 12, and low-dose cytarabine was given subcutaneously twice daily on Days 1 through 14 in escalating doses to a target of 10 mg/m(2) per dose. Of 64 patients who had pathologically confirmed AML, excluding patients with acute promyelocytic leukemia and using World Health Organization criteria, the median age was 71 years, 10 patients (16%) had treatment-related AML, 40 patients (63%) had an antecedent myelodysplastic syndrome or myeloproliferative disorder, and 35 patients (55%) had unfavorable cytogenetics. Thirty-four patients (53%) had an Eastern Cooperative Oncology Group performance status of 2 or 3. RESULTS: Complete remission was achieved in 21 of 61 patients (34%), including 15 of 50 patients (30%) who had secondary or treatment-related AML, 10 of 33 patients (30%) who had unfavorable cytogenetics, and 6 of 34 patients (18%) who had a poor baseline performance status. The mortality rate within the first 4 weeks was 8%. Neutropenic fever was observed in >80% of patients, and 41% of patients had bacteremia. Nonhematologic toxicity generally was mild and reversible and included fatigue, nausea, diarrhea, rash, peripheral edema, and elevated transaminases. There were no clinically significant cardiac arrhythmias. CONCLUSIONS: The addition of arsenic trioxide to low-dose cytarabine appeared to improve responses in elderly patients who had AML compared with either agent alone, and a randomized trial of the combination versus single-agent low-dose cytarabine is ongoing.     

Phase 2 randomized study of p53 antisense oligonucleotide (cenersen) plus idarubicin with or without cytarabine in refractory and relapsed acute myeloid leukemia

BACKGROUND:

The p53 antisense oligonucleotide cenersen has been shown to sensitize acute myeloid leukemia (AML) stem cells to DNA damaging agents.

METHODS:

To determine whether cenersen merits testing in larger efficacy studies, an exploratory study of cenersen in combination with idarubicin either alone or with 1 of 2 doses of cytarabine was performed in first‐salvage AML patients. Patients who either had failed to respond to a single induction course or had responded to induction but relapsed within 12 months were enrolled. Stopping rules based on an expected 14% complete response (CR) rate were applied to each treatment arm.

RESULTS:

Fifty‐three patients were treated, and none of the arms was terminated for lack of activity. Nearly all patients received a single course unless they responded. Ten of the 53 (19%) patients responded (8 CR and 2 CR with incomplete platelet recovery). There was a positive trend for a better response rate with increasing intensity of chemotherapy in the patients refractory to front‐line treatment compared with those who had relapsed previously. One‐third (17/53) of the patients received cenersen inhibitors (acetaminophen and/or high dose antioxidants) during treatment, and none of these responded to treatment. No unique toxicity was attributed to cenersen.

CONCLUSION:

The results of this study suggested that the combination of cenersen with chemotherapy may have clinical efficacy, and additional studies are warranted to explore its full potential. Cancer 2011;. © 2011 American Cancer Society.     

The first case of Philadelphia chromosome-negative acute promyelocytic leukemia following imatinib for chronic myelogenous leukemia

In chronic myelogenous leukemia, chromosomal abnormalities in Philadelphia-negative cells are rare and usually transient, but can infrequently lead to myelodysplastic syndrome and/or acute myeloid leukemia. We report an 82-ear-old patient with an 11-year history of chronic myelogenous leukemia, in complete cytogenetic response, who developed Philadelphia-negative t(15;17)/PMLRARA acute promyelocytic leukemia. This isolated case reaffirms several important clinicopathologic and biologic aspects of chronic myelogenous leukemia, and sheds a unique light on its Philadelphia-negative hematopoiesis. It also underlines the importance of continued cytogenetic monitoring of patients in complete cytogenetic response for the emergence of new chromosomal abnormalities.     

miR -148b 在急性髓系白血病患者及细胞系中的表达与生物学功能

目的：检测 miR -148b 在急性髓系白血病患者和细胞系中的表达水平。初步研究 miR -148b 在急性髓系白血病中的生物学功能。方法：收集急性髓系白血病患者56例,非恶性血液病对照组20例,提取骨髓有核细胞;培养 U937、Kasumi -1、THP -1三种 AML 细胞系。RT - PCR 检测 miR -148b 的表达水平。分别对U937细胞系及 Kasumi -1细胞系转染 miR -148b mimics 和 inhibitor。CCK -8法检测 U937细胞及 Kasumi -1细胞增殖情况,Annexin V/ PI 流式检测细胞凋亡,流式细胞术检测细胞周期。结果：miR -148b 在急性髓系白血病患者及细胞系中较非恶性血液病对照组表达明显下降。U937细胞转染 miR -148b mimics 后,增殖受抑制,细胞凋亡增加,细胞阻滞在 G0- G1期;Kasumi -1细胞转染 miR -148b inhibitor 后,增殖增加,细胞凋亡减少,G0- G1期细胞减少。结论：miR -148b 在急性髓系白血病患者及细胞系中的表达水平均较正常对照显著降低。miR -148b 在急性髓系白血病中起抑癌基因的作用,过表达 miR -148b 可以抑制细胞增殖,促进凋亡。     

Relative and absolute deficiency of bone marrow endogenous colony stimulating activity in acute myeloid leukemia: Relation to response to chemotherapy

The purpose of this study was to determine possible mechanisms for the recently observed association between insensitivity of acute myeloid leukemia (AML) clonogenic cells to colony stimulating activity (CSA) and poor response to induction chemotherapy. The bone marrow endogenous CSA was determined using semi-solid agar cultures by measuring the response of the AML patients' own clonogenic cells to endogenous CSA. The results show that whereas 31% ($\text{5}\text{16}$) of patients at presentation have deficient bone marrow endogenous CSA production over 50% ($\text{11}\text{21}$) have relative deficiency of endogenous CSA, due to insensitivity of the patients' clonogenic cells to CSA.Although there is an association between relative deficiency of endogenous CSA and a poor response to therapy the relationship is not close enough to explain the previously observed highly significant correlation between insensitivity to CSA and poor response to therapy. The results suggest that two independent mechanisms are operative in the association between the CSA-insensitive phenotype and poor response to therapy, one via the tendency to relative endogenous CSA deficiency in the CSA-insensitive group and another via some additional feature of these poor response AML phenotypes which is independent of the presence or absence of endogenous CSA deficiency.     

异基因干细胞移植后急性髓系白血病复发的治疗进展

急性髓系白血病(AML)是一种表型和预后异质性造血干细胞疾病,在接受强化化疗和/或异基因造血干细胞移植(allo-HSCT)的患者中可能得到治愈.随着测序技术的发展揭示了大量分子信息,显著改善了我们对AML基础病理生理学的理解并促进了靶向疗法的发展,目前正在研究几种靶向分子改变的小分子,如FLT3抑制剂、异柠檬酸脱氢酶...     

氧化酚砷诱导急性早幼粒细胞性白血病细胞凋亡的研究

目的：了解氧化酚砷（ｐｈｅｎｙｌａｒｓｉｎｅ ｏｘｉｄｅ,ＰＡＯ）对急性早幼粒细胞性白血病（ＡＰＬ）细胞系ＮＢ４细胞的可能作用。方法：在台盼蓝排除法计数活细胞和细胞活力的基础上,通过细胞形态不和流式细胞仪检测细胞凋亡。通过对细胞进行Ｒｈｏｄａｍｉｎｅ（Ｒｈ）１２３和碘化丙啶双重染色,并应用流式细胞仪检测其荧光强度,以反映细胞线粒体跨膜电位（ΔΨｍ）。结果和结论：低浓度（０．０５和０．０１μｍｏｌ／     

miR-152在急性髓系白血病患者骨髓和细胞系中的表达及生物学功能的初步研究

目的:检测miR-152在急性髓系白血病(AML)患者骨髓和细胞系中的表达水平。初步研究miR-152在急性髓系白血病中的生物学功能。方法:收集急性髓系白血病患者40例,非恶性血液病对照组20例,提取骨髓有核细胞;培养U937、Kasumi-1、THP-1三种急性髓系白血病细胞系。RT-PCR检测miR-152的表达水平。分别对U937细胞系和Kasumi-1细胞系转染miR-152 mimics和inhibitor,CCK-8法检测U937细胞及Kasumi-1细胞增殖情况,Annexin V/PI流式实验检测细胞凋亡,流式细胞术检测细胞周期。结果:miR-152在急性髓系白血病患者骨髓及细胞系中较非恶性血液病对照组表达明显下降。U937细胞系转染miR-152 mimics后,增殖受抑制,细胞凋亡增加,细胞阻滞在G0-G1期;Kasumi-1细胞系转染miR-152 inhibitor后,增殖增加,细胞凋亡减少,G0-G1期细胞减少。结论:miR-152在急性髓系白血病患者骨髓及细胞系中的表达水平均较正常对照组显著降低。miR-152在急性髓系白血病中起抑癌基因的作用,过表达miR-152可以抑制细胞增殖,促进凋亡。     

VEGF inhibition and cytotoxic effect of aplidin in leukemia cell lines and cells from acute myeloid leukemia.

BACKGROUND: Aplidine (APL) is a marine depsipeptide isolated from the Mediterranean tunicate Aplidium albicans that is under clinical phase II development. In contrast to the lack of bone marrow toxicity reported in phase I/II studies, it has been shown to induce cytotoxicity at very low concentration against lymphoblastic leukemia blast, as well as having an impact in the vascular endothelial growth factor (VEGF)/VEGF receptor 1 loop. PATIENTS AND METHODS: To confirm these findings we investigated APL-related VEGF inhibition and its cytotoxic effect on myeloid leukemic cells lines (K-562, HEL and HL60) and fresh leukemia blasts derived from 30 patients with acute myeloid leukemia (AML). The conventional active 4-demetoxi-daunorubicin (idarubicin; IDA) was included as a positive control. RESULTS: APL was found to be significantly (P<0.001) more active than IDA in obtaining 50% growth-inhibition in K-562, HEL and HL60 cell lines. Results obtained with AML blast cells were super imposible. ID(50) ranged from 0.024 to 0.610 microM for IDA (0.200+/-0.176) and from 0.001 to 0.108 microM for APL (0.020+/-0.031). Annexin V tests and cell cycle analysis performed on cell lines confirmed the stronger citotoxic capability of APL as apoptotic inducer and as a G(1) blocker. The inhibitory effects of APL on VEGF release and secretion have been confirmed by ELISA tests performed on HEL: the VEGF concentration in cell surnatant was reduced from 169 to 36 pg/ml after 24 h of exposure to a pharmacological concentration of APL. CONCLUSIONS: APL harbors a strong in vitro antileukemic activity at a concentration achievable in patients at non-myelotoxic doses. Our data also support the notion of an impact on VEGF secretion. Clinical studies with this new marine-derived compound in relapsed/resistant leukemia are underway.     

Early clearance of peripheral blood blasts predicts response to induction chemotherapy in acute myeloid leukemia

BACKGROUND:

Early marrow blast clearance 14 days after induction chemotherapy is an independent prognostic indicator of outcomes in acute myeloid leukemia (AML).

METHODS:

We evaluated the relationship between time to peripheral blood blast clearance after induction and disease status as assessed by day 14 and day 30 marrow biopsies in 162 patients with AML. Day 6 after induction was the optimal cutoff point determined by a receiver operating characteristic analysis and was selected to divide patients into early blast clearance (EBC; ≤6 days; n = 119) and delayed blast clearance (DBC; >6 days; n = 43) groups.

RESULTS:

DBC patients were older, but otherwise the 2 groups were comparable. Marrow blast clearance on day 14 after induction chemotherapy was observed in 84% of patients in the EBC group and 60% in the DBC group. With a median follow‐up of 1538 days, both relapse‐free survival (RFS) (442 vs 202 days, P = .0017) and overall survival (OS) (930 vs 429 days, P < .0001) were longer in the EBC group, and a multivariable analysis showed that EBC independently predicted clearance of marrow blasts at day 14 (P = .0018), remission (P = .0179), RFS (P = .0171), and OS (P = .0122).

CONCLUSIONS:

Early clearance of peripheral blood blasts after induction chemotherapy predicts for early marrow blast clearance, complete remission, RFS, and OS. Cancer 2012. © 2012 American Cancer Society.     

A new set of monoclonal antibodies against acute lymphoblastic leukemia

Six monoclonal antibodies produced by immunization of Balb/c mice with common acute lymphoblastic leukemia (cALL) cells were tested against various types of normal and malignant tissues. ALB1 and ALB2 are directed to the cALL antigen (CALLA gp100); ALB6 recognizes a determinant of p24; ALB7, ALB8 and ALB9 have a pattern of reactivity similar to Ba1. None of these antibodies specifically identify cALL but they should be useful tools for diagnosis or depletion of bone marrow in autologous therapy in transplantation. In addition, the example of ALB6 which acts as a platelet aggregating agent, suggests that the study of other cell systems expressing the antigens associated with cALL may shed light on the function of these antigens and subsequently on the physiopathology of the leukemic cells.     

Prognostic factors and outcome of recurrence in childhood acute myeloid leukemia

BACKGROUND: Outcome after recurrence of childhood acute myeloid leukemia (AML) is poor. We performed this study to identify prognostic factors for recurrence and for survival after recurrence of AML. METHODS: The clinical characteristics, biological features, treatment modalities, and outcomes of children with de novo AML who were enrolled on 3 consecutive clinical protocols from 1987 to 2002 at St. Jude Children's Research Hospital were studied. Regression modeling was used to identify prognostic factors for recurrence and for survival after recurrence. RESULTS: The outcome after recurrence was poor, with a 5-year survival estimate of only 23.3% +/- 5.7%. Multivariable analysis indicated that male sex (P = .005), autologous stem cell transplant before recurrence (P = .097), each additional month from diagnosis to recurrence (P = .041), and stem cell transplant after recurrence (P < .001) were associated with a better survival after recurrence, whereas M5 or M7 morphology (P = .001) were significantly predictive of a lower survival estimate after recurrence. CONCLUSIONS: Survival after recurrence was poor in children with AML. Novel therapies are urgently needed to prevent or to treat recurring AML.     

LD-HA方案在急性髓系白血病治疗中的应用

上世纪80年代,由于缺乏有效的支持治疗（如抗生素、输血、细胞因子等）,小剂量化疗药物治疗急性髓系白血病（acute myeloid leukemia,AML）成为当时的一个研究热点,特别是小剂量阿糖胞苷化疗方案的应用,取得了良好效果。1984年Body等体外证明了小剂量三尖杉酯碱有诱导HL-60细胞分化成熟的作用。1985年黄昌亮等国内外率先报告了LD-HA方案治疗AML的疗效,目前已成为不适合标准治疗方案的急性髓系白血病患者主要选择性方案之一,本文就LD-HA临床应用、疗效及存在问题作一综述。     

Surface glycoproteins of human non-T, non-B acute lymphocytic leukemia cell lines

The surface glycoproteins of 4 human acute lymphocytic leukemia (ALL) cell lines with immunologic surface marker profiles suggestive of a non-T, non-B lymphocyte derivation (absence of surface immunoglobulin, complement and Fc receptors presence of non-T, non-B common ALL (cALL) and Ia-like antigen) have been analyzed by the galactose oxidase tritiated sodium borohydride labelling technique. The surface glycoprotein patterns have been compared with those of normal B-, T-, O-blood lymphocytes and with fresh ALL and chronic lymphocytic leukemia cells (CLL). All cALL lines express GP 42, GP 31 and GP 24, which have been identified as HLA, and Ia-like antigens, respectively. With one exception (the KM3 line) the cALL lines have GP 120 and GP 130 as major surface GPs a feature shared with fresh ALL and acute myelocytic leukemia cells. Two of the lines (NALL-1 and Nalm-1) also express a GP 210 K found as a major band on O- and B-lymphocytes and on fresh ALL cells. The dominating surface GP on KM 3 has an apparent mol. wt. of 1000,000 Daltons which is not clearly seen on other cells examined. The analyses suggest a common surface glycoprotein pattern of cALL (GP 210, GP 130, GP 120, GP 42, GP 31 and GP 24) but also re-emphasizes that cALL, as defined by immunologic surface marker analysis, is heterogenous neoplasia as the major surface GPs in KM 3 and Nalm-16 differ from those of the other cALL lines and the fresh cALL cells.     

急性髓性白血病患者LRP、MRP基因表达

目的：研究急性髓性白血病ＬＲＰ基因、ＭＲＰ基因表达与疗效的关系，方法：用ＲＴ－ＰＣＲ法检测３７例急性髓性白血病ＬＲＰ基因、ＭＲＰ基因表达，按此两种基因表达分成ＬＲＰ－／ＭＲＰ－、ＬＲＰ－ＭＲＰ＋、ＬＲＰ＋／ＭＲＰ－、ＬＲＰ＋／ＭＲＰ＋共４组，统计比较各组疗效。结果：与ＬＲＰ－／ＭＲＰ－组比较，其余３组疗效较低，其中ＬＲＰ＋／ＭＲＰ－组、ＬＲＰ＋／ＭＲＰ＋组存在显著差异，但ＬＲＰ＋／ＭＲＰ＋组与ＬＲ     

沉默SIRT2基因对急性粒细胞白血病细胞增殖的影响

目的 SIRT2是组蛋白去乙酰化酶家族成员之一,其参与多种肿瘤的发生和发展.本研究旨在探讨SIRT2基因在人急性粒细胞白血病细胞NB4和耐药细胞HL60/A增殖中的作用,分析SIRT2对于细胞周期的影响,寻找治疗急性粒细胞白血病的新靶点.方法 蛋白质印迹法检测SIRT2在急性粒细胞白血病细胞HL60/A和NB4中的表达,针对SIRT2 mRNA不同的靶序列设计并合成shRNA,构建真核表达干扰载体,制备慢病毒并感染HL60/A和NB4细胞,用嘌呤霉素筛选出稳定表达shRNA的细胞,应用蛋白质印迹法验证干扰效率;采用改良MTT法检测沉默SIRT2基因前后HL60/A和NB4细胞增殖的改变;用流式细胞术检测沉默SIRT2基因前后细胞周期的变化.结果 在HL60/A和NB4细胞中SIRT2蛋白表达水平均增高,F=43.22,P=0.002.用慢病毒感染HL60/A(F=184.9,P＜0.001)和NB4(F=354.7,P＜0.001)后,SIRT2基因的蛋白表达水平显著降低.MTT结果显示,与对照组细胞比较,48 h沉默SIRT2基因的HL60/A(F=208.4,P＜0.001)和NB4(F=27.58,P＜0.001)细胞的生长速度变慢;于72 h这种抑制作用更加明显,其中HL60/A的增殖速度明显变慢,F=555.9,P＜0.001.流式细胞术检测细胞周期结果显示,G0～G1期百分比,与对照组HL60/A/con(27.62±1.01)％相比,干扰组HL60/A/sh1 (37.14±0.03)％(F=7.61,P＜0.001)和HL60/sh2(37.51±3.63)％(F=3.89,P=0.009)比例显著增加;与对照组NB4/con(27.49±0.50)％相比,干扰组NB4/sh1(38.26±1.93)％(F=8.02,P=0.007)和NB4/sh2(35.23±1.11)％(F=3.50,P＜0.001)比例显著增加;S期细胞百分比,与对照组HL60/A/con(51.94±1.15)％相比,干扰组HL60/A/sh1 (40.24±0.66)％(F=3.34,P＜0.001)和HL60/sh2(42.73±2.76)％(F=10.33,P=0.004)比例显著降低;与对照组NB4/con(50.65±0.23)％相比,干扰组NB4/sh1(39.58±1.11)％(F=3.02,P＜0.001)和NB4/sh2(43.82±1.33)％(F=10.42,P=0.011)比例显著降低;G2～M期细胞比例无明显变化.结论 干扰SIRT2基因的表达可使细胞周期阻滞在G0～G1期,从而抑制急性粒细胞白血病细胞HL60/A和NB4的增殖.