混合造血干细胞移植联合DLI-IL-2治疗急性髓性白血病的疗效

目的:探讨急性髓性白血病(acute myelogenous leukemia,AML)患者采用自体外周血干细胞混合HLA半相合异体骨髓移植(autologous peripheral blood stem cell mixed with HLA haploidentical allogeneic bone marrow transplantation,Mixed-HSCT)联合供体淋巴细胞输注+白介素2(donor lymphocyte infusion combined interleukin-2,DLI-IL-2)治疗的疗效。方法:采用联合治疗方案的试验组23例AML患者中男性15例、女性8例,中位年龄22(17～41)岁;采用单纯移植治疗的对照组14例AML患者中男性10例、女性4例,中位年龄21(19～40)岁。两组患者在完全缓解期采用TBI+VEMAC方案预处理,对照组患者接受单纯Mixed-HSCT移植,试验组患者接受Mixed-HSCT且造血重建后继续DLI-IL-2治疗1～8次;各组在治疗前后进行染色体核型分析及骨髓检查。随访时间>3年。结果:两组患者均获得造血重建,无移植物抗宿主病(graft-versus-host disease,GVHD)发生。试验组有6例形成混合嵌合体(46XX/46XY),随访显示存活15例,长期无病存活率(disease-free survival,DFS)为65.2%;对照组有3例形成混合嵌合体(46XX/46XY),随访显示存活7例,DFS为50.0%。两组患者治疗后的不良反应(口腔溃疡、出血性膀胱炎、发热等)相似。结论:Mixed-HSCT联合DLI-IL-2治疗对AML患者长期无病生存有积极意义,无严重不良反应。     

急性髓性白血病混合造血干细胞移植后DLI＋IL-2治疗的临床研究

目的：探讨急性髓性白血病（acute myeloid leukemia,AML）患者在接受自体外周血干细胞混合人类白细胞抗原（human leukocyte antigen,HLA）半相合异体骨髓移植（Mixed-HSCT）后,继予供体淋巴细胞输注＋白介素2（DLI＋IL-2）治疗的疗效。方法：对23例AML患者在完全缓解期采用TBI＋VEMAC预处理方案,实施Mixed-HSCT。造血恢复后给予DLI＋IL-2治疗1-8次。结果：所有患者均获得造血重建,中性粒细胞（ANC）≧0.5×109/L的中位时间为14（12-17）天,白细胞（WBC）≧4.0×108/L的中位时间为17（16-21）天。血小板（PLT）≧20×108/L的中位时间为21（19-23）天,PLT≧50×108/L的中位时间为25（24-27）天。＋16至＋21天时骨髓检查示恢复期骨髓象,无移植物抗宿主病（graft versus host disease,GVHD）发生,有6例形成混合嵌合体（46XX/46XY）。经过3年以上随访,存活15例,长期无病存活率（DFS）为65.2%。结论：Mixed-HSCT后应用DLI＋IL-2治疗对急性髓性白血病患者的长期无病生存有积极意义。     

急性髓性白血病混合造血干细胞移植后DLI+IL-2治疗的临床研究

目的:探讨急性髓性白血病(acute myeloid leukemia,AML)患者在接受自体外周血干细胞混合人类白细胞抗原(human leukocyte antigen,HLA)半相合异体骨髓移植(Mixed-HSCT)后,继予供体淋巴细胞输注+白介素2(DLI+IL-2)治疗的疗效。方法:对23例AML患者在完全缓解期采用TBI+VEMAC预处理方案,实施Mixed-HSCT。造血恢复后给予DLI+IL-2治疗1-8次。结果:所有患者均获得造血重建,中性粒细胞(ANC)≧0.5×109/L的中位时间为14(12-17)天,白细胞(WBC)≧4.0×108/L的中位时间为17(16-21)天。血小板(PLT)≧20×108/L的中位时间为21(19-23)天,PLT≧50×108/L的中位时间为25(24-27)天。+16至+21天时骨髓检查示恢复期骨髓象,无移植物抗宿主病(graft versus host disease,GVHD)发生,有6例形成混合嵌合体(46XX/46XY)。经过3年以上随访,存活15例,长期无病存活率(DFS)为65.2%。结论:Mixed-HSCT后应用DLI+IL-2治疗对急性髓性白血病患者的长期无病生存有积极意义。     

评分法研究复发急性白血病再生耐药现象

目的：建立评价急性髓性白血病（ＡＭＬ）再生耐药的方法。材料和方法：用白血病祖细胞培养（ＣＦＵ－Ｌ）,ＭＴＴ药物敏感试验,细胞周期中Ｓ期 细胞百分比（Ｓ％）,细胞内柔红霉素（ＤＮＲ）测定其荧光指数（ＦＩ）和耐药指数（ＤＲＩ）,ＡＰＡＡＰ法测Ｐ－１７０糖蛋白表达以及半定量ＲＴ－ＰＧＲ检测Ｂcl-ＸＬmＲ表达丰度,评价ＡＭＬ细胞的药物敏感度和增殖情况。首先选取初治ＡＭＬ病例研究评分指标与细胞耐药和细胞生长的     

Mutation of the TP53 gene in acute lymphoblastic leukemia does not affect survival outcomes after haploidentical hematopoietic stem cell transplantation

Previous studies have demonstrated that TP53 mutation is correlated with insufficient therapy response and unfavorable prognosis in acute lymphoblastic leukemia (ALL). Few studies have investigated the impact of TP53 mutation in ALL patients after haploidentical hematopoietic stem cell transplantation (haplo-HSCT). We completed a retrospective study of 65 ALL patients with available TP53 status who underwent haplo-HSCT. They were divided into a TP53 mutation group (TP53^mut) and a TP53 wild-type (TP53^wt) group. TP53^mut showed comparable 2-year cumulative incidence of relapse (CIR) rates (13.1% vs 12.5%, P = .96) and 2-year leukemia-free survival (LFS) (74.2% vs 77.4%, P = .80) with TP53^wt. No significant differences in 2-year overall survival (OS) rates (82.9% vs 87.3%, P = .61) or 2-year NRM rates (12.7% vs 10.2%, P = .69) were observed in TP53^mut and TP53^wt patients. Multivariate analysis suggested that white blood cell (WBC) count at initial diagnosis (>50 × 10⁹/L: hazard ratio [HR] = 3.860, P = .016) and age (>40 years old: HR = 4.120, P = .012) are independent risk factors for 2-year LFS. Our study showed that TP53 mutations may not be related to the unfavorable impact on survival in ALL patients after treatment with haplo-HSCT. The present results suggested that haplo-HSCT may eliminate the poor prognosis effect of TP53 mutation in ALL.     

联合克拉屈滨预处理方案在难治/复发性急性髓系白血病患者异基因造血干细胞移植中的疗效观察

目的:评价联合克拉屈滨预处理方案在异基因造血干细胞移植(allo-HSCT)治疗难治/复发性急性髓系白血病(AML)中的疗效.方法:收集并分析2017年4月至2019年10月在我院层流病房行联合克拉屈滨预处理方案并行异基因造血干细胞移植治疗的17例难治/复发性AML患者的临床资料.结果:17例患者均完成造血重建,粒细胞...     

Microvascular endothelial cells increase proliferation and inhibit apoptosis of native human acute myelogenous leukemia blasts

Interactions between acute myelogenous leukemia (AML) blasts and neighbouring endothelial cells in the bone marrow seem important both for disease development and susceptibility to chemotherapy. We investigated the effects of soluble mediators released by microvascular endothelial cells on native human AML cells. AML cells derived from 33 patients were cocultured with microvascular endothelial cells, separated by a semipermeable membrane. We investigated the effect of coculture on AML cell proliferation, viability/apoptosis and cytokine release. Coculture increased AML cell proliferation, and this growth enhancement included the clonogenic leukemia cell subset. Increased release of several soluble mediators was also detected (interleukin 3, interleukin 6, granulocyte-macrophage and granulocyte colony-stimulating factors) in cocultures. Our cytokine neutralization experiments suggest that an intercellular crosstalk involving several soluble mediators contribute to the increased leukemia cell proliferation. The presence of endothelial cells had an additional antiapoptotic effect on the AML cells. The endothelial cells did not have any growth-enhancing effect on native human acute lymphoblastic leukemia cells. Our in vitro results suggest that the release of soluble mediators by microvascular endothelial cells supports leukemic hematopoiesis through paracrine mechanisms by direct enhancement of AML blast proliferation and by inhibition of leukemic cell apoptosis.     

Sialyltransferase activity in leukemia

Sialyltransferase activity in blasts from acute lymphoblastic leukemia (ALL) was markedly lower (1.68 ± 1.23 pmol/mg protein) than those (6.18 ± 2.22 pmol/mg protein) of lymphocytes from normal volunteers (t < 0.001). On the contrary, enzyme activity was significantly increased in blasts (1.21 ± 0.38 pmol/mg protein) from acute non-lymphoblastic leukemia, compared to the level (0.53 ± 0.32 pmol/mg protein) of mature granulocytes (t < 0.001). In TdT-negative CML in blast crisis, sialyltransferase activity (2.11 ± 0.88 pmol/mg protein) was significantly higher than those of mature granulocytes (t < 0.001), whereas no significant difference in the enzyme activity was noted between the blasts from TdT-positive CML in blast crisis and from ALL. In TdT-positive ALL cases, there was an inverse relationship (r = ?0.85, t < 0.01) between sialyltransferase activity and terminal deoxynucleotidyl transferase (TdT) activity of the blasts. Therefore, sialyltransferase in leukocytes may be a unique enzyme in which changes in activity relate to the differentiation or malignant transformation of leukocytes.     

异基因造血干细胞移植治疗急性T 淋巴细胞白血病和淋巴瘤：附50例临床报告

目的 探讨异基因造血干细胞移植（allogeneic hematopoietic stem cell transplantation,allo-HSCT）治疗急性T淋巴细胞白血病（T cell acute lymphoblastic leukemia,T-ALL）和急性T淋巴母细胞淋巴瘤（T cell acute lymphoblastic lymphoma, T-LBL）的疗效及预后。方法 回顾性分析2014—2019年于航天中心医院接受allo-HSCT的50例T-ALL/LBL患者的临床资料,分析其临床疗效、并发症及预后。结果 50例患者中男性41例,女性9例,中位年龄20.5岁（范围：9.0~63.0岁）;单倍体移植44例,脐血移植 2例,同胞全合移植 4例;T-ALL 40例,T-LBL 10例;移植前处于完全缓解（CR）状态16例,处于未完全缓解（非CR）状态34例。移植后,中位随访20个月（范围：1~84个月）,存活23例,死亡27例;移植后24个月的总生存率和无复发生存率分别为50.0%和44.0%,36个月的总生存率和无复发生存率分别为45.5%和40.0%。随访期间,共有20例患者复发,复发率为40.0%（20/50）。移植前获CR、无髓外病变、无中枢神经系统受累的患者预后较好,而移植前有无基因突变、不同预处理方案、有无急性/慢性GVHD患者的总生存期及无复发生存期组间比较差异无统计学意义（均P>0.05）。结论 在这项小样本、无对照的临床研究中,T-ALL/LBL患者在缓解期行allo-HSCT可能较挽救性移植的生存预后有所改善,其中复发为移植失败的主要原因。     

Haploidentical hematopoietic stem cell transplantation in adults with Philadelphia‐negative acute lymphoblastic leukemia: No difference in the high‐ and low‐risk groups

Allogeneic hematopoietic stem cell transplantation (HSCT) is the most effective post‐consolidation therapy and curative option for adult patients with Philadelphia chromosome‐negative (Ph‐negative) acute lymphoblastic leukemia (ALL) in first complete remission (CR1). A human leukocyte antigen (HLA)‐haploidentical related donor (haplo‐RD) is one of the most important alternative sources for those without HLA‐identical sibling donor (ISD). The present study aimed to evaluate the outcomes of haploidentical hematopoietic stem cell transplantation (haplo‐HSCT) in adult Ph‐negative ALL CR1 patients (n = 183). We produced an unmanipulated haplo‐HSCT protocol including granulocyte colony stimulating factor (G‐CSF) for all donors, intensive immune suppression, anti‐thymocyte globulin, and combination of G‐CSF‐primed bone marrow harvest and G‐CSF‐mobilized peripheral blood stem cells harvest as the source of stem cell grafts. The median age for high‐risk versus low‐risk groups were 29 versus 23 years. Three‐year incidences of relapse mortality and nonrelapse mortality for high‐risk versus low‐risk groups were 7.1% versus 11.1% (p = 0.498) and 18.0% versus 16.2% (p = 0.717), respectively. Three‐year probabilities of disease‐free survival and overall survival for high‐risk versus low‐risk groups were 67.6% versus 68.2% (p = 0.896) and 74.9% versus 72.7% (p = 0.981), respectively. Multivariate analysis showed that limited cGVHD and a lower pre‐HSCT comorbidity burden were associated with better outcomes. In summary, comparable outcomes were observed among high‐ and low‐risk Ph‐negative ALL CR1 patients after haplo‐HSCT. Haplo‐RD could be considered for adults with Ph‐negative ALL in CR1 as an important alternative source of donors in cases when no ISD is available.     

Therapy‐related acute myelogenous leukemia and myelodysplastic syndrome in patients with acute lymphoblastic leukemia treated with the hyperfractionated cyclophosphamide,vincristine, doxorubicin,and dexamethasone regimens

BACKGROUND:

Secondary malignancies including myeloid neoplasms occur infrequently in acute lymphoblastic leukemia (ALL) and to the authors' knowledge have not been as well documented in adults as in children.

METHODS:

A total of 641 patients with de novo ALL who were treated with the hyper‐CVAD (hyperfractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone) regimen or its variants were analyzed.

RESULTS:

Sixteen patients (2.49%) developed secondary acute myelogenous leukemia (AML) (6 patients) or myelodysplastic syndrome (MDS) (10 patients). At the time of ALL diagnosis, the median age was 53 years; cytogenetics were normal in 11 patients, pseudo‐diploidy with del(2) in 1 patient, t(9;22) in 1 patient, and unavailable in 3 patients. Frontline therapy included hyper‐CVAD in 7 patients, hyper‐CVAD with rituximab in 8 patients, and hyper‐CVAD with imatinib in 1 patient. Karyotype at time of AML/MDS diagnosis was ?5, ?7 in 9 patients, normal in 1 patient, complex in 1 patient, inv(11) in 1 patient, t(4;11) in 1 patient, del(20) in 1 patient, and unavailable in 2 patients. Secondary AML/MDS developed at a median of 32 months after ALL diagnosis. Cytarabine plus anthracycline–based treatment was given to 12 patients with AML and high‐risk MDS. One patient with MDS received arsenic trioxide, 1 received clofarabine, and 2 received decitabine. Response to treatment was complete remission in 3 patients, partial remission in 6 patients, and no response in 6 patients; 1 patient was untreated. Eight patients (1 with AML and 7 with MDS) underwent allogeneic stem cell transplantation, and all but 2 died at a median of 3 months (range, 0.5–11 months) after transplantation. The median overall survival after a diagnosis of secondary AML and MDS was 9.25 months (range, 1+ to 26+ months).

CONCLUSIONS:

Secondary AML and MDS occur infrequently in adult patients with de novo ALL treated with the hyper‐CVAD regimens, and response to therapy is poor. Cancer 2009. © 2008 American Cancer Society.     

Low leukemia burden improves blinatumomab efficacy in patients with relapsed/refractory B-cell acute lymphoblastic leukemia

Background

A lower baseline bone marrow blast percentage (bBMB%) is associated with better outcomes in patients with B-cell acute lymphoblastic leukemia (B-ALL) receiving blinatumomab. The objective of this analysis was to investigate the association between bBMB% and treatment outcomes in relapsed/refractory (R/R) B-ALL.

Methods

Data from five trials of blinatumomab for R/R B-ALL were pooled for analyses. Patients were placed in one of three groups: group 1, ≥50% bBMBs; group 2, ≥25% to <50% bBMBs; group 3, ≥5% to <25% bBMBs. Response and survival outcomes were compared between groups.

Results

Data from 683 patients (166 pediatric, 517 adult) were analyzed. Collectively, patients in groups 2 and 3 had significantly higher odds of achieving a complete remission (CR) (odds ratio [OR], 3.50 [95% confidence interval (CI), 2.23–5.48] and 3.93 [95% CI, 2.50–6.18], respectively; p < .001) and minimal/measurable residual disease response (OR, 2.61 and 3.37, respectively; p < .001) when compared with group 1 (reference). Groups 2 and 3 had a 37% and 46% reduction in the risk of death (hazard ratio [HR], 0.63 and 0.54, respectively; p < .001) and a 41% and 43% reduction in the risk of an event (relapse or death) (HR, 0.59 and 0.57, respectively; p < .001) compared with group 1. No significant differences in response or survival outcomes were observed between groups 2 and 3. Seven of nine patients whose bBMB% was lowered to <50% with dexamethasone achieved CR with blinatumomab.

Conclusion

Any bBMB% <50% was associated with improved efficacy following blinatumomab treatment for R/R B-ALL.     

PTEN基因对急性淋巴细胞白血病细胞增殖、侵袭的影响

目的:观察第10号染色体同源缺失性磷酸酶-张力蛋白同源的基因（PTEN）对急性淋巴细胞白血病细胞增殖、侵袭的影响。方法:在人急性淋巴细胞白血病细胞CEM-C1中转染PTEN过表达载体（pBP-PTEN）,同时转染对照载体（pBP）,以没有转染的细胞作为对照,RT-PCR和Western blot检测转染后细胞中PTEN的水平;MTT和集落形成试验检测细胞增殖能力;Transwell小室检测细胞侵袭和迁移能力;Western blot检测PTEN、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）、p38丝裂原活化蛋白激酶（p38MAPK）、磷酸化的p38MAPK（p-p38MAPK）的水平。结果:转染对照载体的CEM-C1细胞中PTEN水平、细胞光密度（OD）值、克隆形成数目、侵袭数目、迁移数目及PTEN、MMP-2、MMP-9、p38MAPK、p-p38MAPK水平与未转染细胞相比没有明显变化（P＞0.05）。转染PTEN过表达载体后的CEM-C1细胞中PTEN水平和p-p38MAPK水平升高,细胞OD值、克隆形成数目、侵袭数目、迁移数目和MMP-2、MMP-9水平降低,与未转染细胞相比差异具有统计学意义（P＜0.05）。结论:PTEN抑制急性淋巴细胞白血病细胞增殖、侵袭及迁移,促进p38MAPK信号通路激活。     

二硫化二砷对维甲酸耐药急性早幼粒细胞白血病细胞株NB4-R2增殖和凋亡的影响

目的:研究As2S2对维甲酸耐药急性早幼粒细胞白血病细胞株NB4-R2增殖和凋亡的影响,并探讨其机制。方法:CCK-8法检测不同浓度As2S2作用于NB4-R2细胞24、48和72h对细胞的生长抑制率,计算半数抑制浓度（IC50）,Annexin V-FITC/PI双染法检测As2S2对NB4-R2细胞凋亡的影响,实时定量PCR（quantitative real-time PCR,qRT-PCR）和Western blot检测As2S2作用前后NB4-R2细胞PML-RARα融合基因和融合蛋白的表达水平。结果:不同浓度As2S2分别作用于NB4-R2细胞24、48和72h后,细胞增殖均受到不同程度抑制,且该抑制作用呈时间和剂量依赖性,As2S2对NB4-R2细胞的IC50分别为（17.64±2.11）μmol/L、（8.57±1.03）μmol/L和（5.82±0.71）μmol/L。流式细胞术结果表明As2S2处理24、48和72h后细胞凋亡率分别为正常对照组的（4.43±0.12）倍、（6.50±0.09）倍和（8.63±0.36）倍。qRT-PCR结果表明PML-RARα mRNA表达水平较作用前分别降低了（1.47±0.09）倍、（2.86±0.82）倍和（7.46±1.09）倍,同时,PML-RARα融合蛋白表达水平较作用前分别降低了（1.36±0.16）倍、（1.85±0.33）倍和（3.99±0.63）倍（P<0.05）。结论:As2S2可显著抑制NB4-R2细胞增殖并促进细胞凋亡,其机制与诱导PML-RARα融合基因和融合蛋白降解密切相关。     

Methods of dendritic cell preparation for acute lymphoblastic leukemia immunotherapy in children

Cell immunotherapy through dendritic cells (DC) presents a hopeful strategy for the treatment of various tumors. The aim of our study was to find which progenitor cells are most suitable for the preparation of dendritic cells in acute lymphoblastic leukemia (ALL) in pediatric patients, whether blasts from bone marrow or dendritic cells generated from peripheral blood mononuclear cells taken at the time of remission after induction chemotherapy. DC generated from the BM blasts of patients with B-ALL and T-ALL (n=15) at the time of diagnosis expressed low levels of costimulatory molecules and CD markers typical for mature DC. In contrast, DC cultivated from peripheral mononuclear cells of patients (n=9) had comparable morphology and expression of costimulatory molecules to DC obtained from healthy individuals, which was even higher after tumor lysate pulsing. Autologous lymphocyte proliferation increased after DC blasts lysate pulsation and further after lymphocyte restimulation, showing evidence of induction of specific cytotoxic lymphocytes. When comparing both cell sources for the preparation of DC in patients with ALL, it appears that peripheral mononuclear cells obtained after chemotherapy are more suitable than bone marrow leukemic blasts due to similar morphology, phenotypic, and functional capacity to monocytes of healthy donors. Despite this, it is necessary to take into account individual variability when preparing DC-based vaccines. The final verification of the efficiency of immunotherapy against residual hematopoietic malignant cells in patients with ALL can only be obtained through a clinical study.     

异基因造血干细胞移植治疗白血病

目的探讨异基因造血干细胞移植(Allo-HSCT)治疗白血病的疗效、造血重建及生存情况.方法白血病患者10例,其中同胞间HLA相合的异基因外周血干细胞移植(Allo-PBSCT)7例,无亲缘关系HLA不全相合脐血移植(UCBT)3例.结果9/10例受者获造血重建,UCBT患者造血重建速度较HLA相合的同胞PBSCT慢,1例UCBT移植后35天造血未重建,回输自体外周血干细胞后,仍未能重建造血,于72天死亡.其余至今均无病生存.结论Allo-HSCT是目前治愈白血病的有效方法,对于无同胞HLA相合的供者,选择细胞数量较高、HLA 1～2个位点不合的UCBT仍然有效可行.     

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。     

ABT-199体外对急性T淋巴细胞白血病细胞株Molt4细胞增殖凋亡影响及其机制研究

目的 大多数肿瘤细胞对化疗诱导细胞凋亡的耐受与Bcl-2家族蛋白有关,ABT-199是针对抗凋亡蛋白Bcl-2的选择性拮抗剂.本研究旨在探讨ABT-199体外对急性T淋巴细胞白血病Molt4细胞株增殖凋亡的影响及其机制.方法 体外培养Molt4细胞,CCK8法检测不同浓度ABT-199对Moh4的增殖抑制作用,DAPI细胞核染色后荧光显微镜观察细胞凋亡特征,Annexin Ⅴ/PI双染法检测不同浓度ABT-199对Molt4细胞的诱导凋亡作用,JC-1染色法检测不同浓度ABT-199作用于Moh4细胞后线粒体膜电位的变化,蛋白质印迹法检测经ABT-199处理后线粒体信号通路相关蛋白(Bcl-2、PARP和cleaved Caspase-3蛋白)表达水平的变化.结果 ABT-199对Molt4细胞具有抑制增殖的作用,48 h的IC50值为(4.63±0.15) μmol/L.荧光显微镜观察显示,ABT-199处理Moh4细胞后细胞核染色质染色浓聚,细胞核碎裂,出现凋亡小体,且随着药物浓度的增加上述形态变化更加明显;Annexin Ⅴ/PI检测细胞凋亡结果显示,0、1、2、4、8 μmol/L的ABT-199诱导细胞凋亡的百分比分别为(3.09±1.16)％、(11.59±5.58)％、(25.37±7.42)％、(46.38±7.05)％和(80.60±11.18)％,给药组与对照组的凋亡比例差异有统计学意义,x2=18.286,v=4,P=0.001.JC-1检测结果显示,ABT-199能促使Molt4细胞的线粒体膜电位下降,呈浓度依赖性,给药组与对照组的线粒体膜电位下降比例差异有统计学意义,x2 =17.386,v=4,P=0.002.蛋白质印迹法检测结果显示,给药组线粒体通路中Bcl-2表达水平显著下降(x2=9.024,v=3,P=0.029),并出现Caspase-3下游底物多聚ADP核糖聚合酶(poly ADP ribose poly-merase,PARP)的裂解,同时出现了Caspase-3裂解片段的累积.结论 ABT-199在体外能抑制急性T淋巴细胞白血病Molt4细胞的增殖及诱导其凋亡,其机制与激活线粒体信号通路相关.