Racemic Etodolac is cytotoxic and cytostatic for B-cell precursor acute lymphoblastic leukemia cells

Several epidemiological studies have provided evidence that administration of nonsteroidal anti-inflammatory drugs (NSAIDs) could have a prophylactic effect against some cancers such as sporadic colorectal cancer and leukemia. Indeed, various NSAIDs have been shown to induce apoptosis in malignant cells. We evaluated the effect of racemic Etodolac on proliferation and cell survival in B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cells. Etodolac decreased survival of Nalm-16 and Nalm-6 BCP-ALL cell lines and also decreased cell proliferation in Nalm-16 cell line. Ours findings indicate, for the first time to our knowledge, that Etodolac is cytotoxic and cytostatic for BCP-ALL cells.     

Wnt信号通路与卵巢癌治疗新进展

Wnt通路是一条高度保守的传导通路,Wnt信号通路异常激活与肿瘤发生发展有密切关系。近年来,不少研究证明卵巢癌中存在异常激活的Wnt通路信号蛋白。该文复习了卵巢癌与Wnt通路相关的新近研究进展,对Wnt信号通路、其在卵巢癌发生发展的作用及Wnt通路中潜在的卵巢癌治疗新靶点做一综述,旨在为卵巢癌的临床治疗提供新思路。     

Hippo信号通路核心元件MST1在急性白血病患者中的表达及意义

本研究旨在探讨Hippo信号通路核心元件MST1基因在急性白血病患者中的表达情况及其与白血病发生、发展的关系。采用实时定量PCR检测50例初治急性白血病患者、33例正常人、23例完全缓解缓解患者及12例难治/复发患者的MST1基因的表达情况,采用Western blot进一步验证其蛋白的表达水平,结合临床资料对患者的预后因素进行分析。结果表明,与正常人相比,急性白血病初治患者MST1基因的表达水平明显减低(P<0.05),完全缓解前后差异有统计学意义(P<0.05),难治/复发患者与初治患者MST1基因表达水平无明显差异(P>0.05),完全缓解患者与正常人MST1表达水平无明显差异(P>0.05)。结论:MST1基因的低表达与急性白血病的发病及预后相关。     

C-MYC siRNA对急性淋巴细胞白血病Jurkat细胞凋亡的诱导作用

本研究探讨C-MYC siRNA对急性淋巴细胞白血病细胞系Jurkat细胞的增殖、凋亡的影响.采取化学合成法合成针对C-MYC rnRNA的第1545-1565靶位点设计的siRNA,经转染剂(HiPerFect transfection reagent)转染入Jurkat细胞,观察细胞形态学变化;应用四唑氮化合物(MTS)法绘制细胞生长曲线;用细胞集落培养观察C-MYC siRNA对Jurkat细胞增殖的影响;应用流式细胞术和TdT酶介导的末端缺失原位标记(TUNEL)法分析细胞的凋亡.结果表明,不同浓度C-MYC siRNA作用于Jurkat细胞后,细胞的生长受到不同程度的抑制,抑制率随C-MYC siRNA浓度的增高而增高.C-MYC siRNA对集落形成也有明显的抑制作用.TUNEL法、流式细胞仪均检测出细胞凋亡,且随着作用时间的延长,其凋亡率也逐渐上升.结论:化学合成法合成的C-MYC siRNA能明显抑制Jurkat细胞的增殖与集落形成,并可诱导 Jurkat细胞发生凋亡.     

辛伐他汀对人急性单核细胞白血病SHI-1细胞PI3K/AKT通路的影响

本研究探讨羟甲基戊二酸单酰辅酶A还原酶抑制剂辛伐他汀对人急性单核细胞白血病株(SHI-1)细胞增殖和凋亡的影响及PI3K/AKT通路变化。取对数生长期细胞,实验分为阴性对照组和辛伐他汀处理组(终浓度分别为5、10、15μmol/L),培养24、48、72 h。采用MTT法观察SHI-1细胞增殖能力;流式细胞术测定SHI-1细胞凋亡指标变化;PCR芯片研究SHI-1细胞PI3K/AKT通路84个特异性基因mRNA的差异表达。结果表明,辛伐他汀对SHI-1细胞有明显抑制增殖和促凋亡作用,呈时间与剂量依赖性。15μmol/L辛伐他汀处理SHI-1细胞24、48、72h,细胞增殖抑制率分别为26.82%、47.09%、63.92%,细胞早期凋亡率分别为5.73%、13.25%、15.59%。与对照组相比,15μmol/L辛伐他汀处理SHI-1细胞48 h组中有39个基因表达发生改变,其中26个基因表达下调、13个基因表达上调。结论:辛伐他汀能抑制SHI-1细胞增殖并诱导其凋亡,其诱导凋亡机制可能与辛伐他汀调节PI3K/AKT通路相关基因的表达有关。     

Targeting the interaction of Aurora kinases and SIRT1 mediated by Wnt signaling pathway in colorectal cancer: A critical review

The Aurora kinases belong to the family of serine/threonine kinase, a central regulator of mitosis and their expression increased during G2/M phase. It is classified into Aurora A, B and C, each has distinct roles in cellular processes, which includes regulation of spindle assembly, function of centrosomes, cytoskeleton and cytokinesis. During cancer growth, their rapid increase makes most attractive marker for cancer treatment at present. However Aurora A kinase is known to be a marker for cancer therapy, the most important serine/threonine kinase of Aurora B kinase involvement in cancer is still inadequate. Subsequently, the recent findings revealed that the class III histone deacetylase of SIRT1 is a key regulator to activate Aurora kinases from S phase damaged DNA through Wnt signaling pathway. Even if both Aurora A kinase and SIRT1 serve as a marker for cancer therapy, the present review reveals it is interaction in Wnt signaling pathway that solely for colorectal cancer.     

Extracellular matrix protein ITGBL1 promotes ovarian cancer cell migration and adhesion through Wnt/PCP signaling and FAK/SRC pathway

Despite the advances in cancer treatment and the progresses in tumor biological, ovarian cancer remains a bad situation. In current study, we found a novel extracellular matrix protein, ITGBL1, which is highly expressed in ovarian cancer tissues by immunohistochemistry examination. The expression pattern of ITGBL1 in malignant tissues inspired us to investigate its role in ovarian cancer progression. Both loss- and gain-function assays revealed that ITGBL1 could promote ovarian cancer cell migration and adhesion. As it’s a secreted protein, we further used recombinant ITGBL1 protein treated cancer cells and found that ITGBL1 promotes cell migration and adhesion in a concentration dependent manner. Furthermore, we found that ITGBL1 not only influences the activity of Wnt/PCP signaling but also affects FAK/src pathway in vitro. Taken together, our results suggest that highly expressed ITGBL1 could promotes cancer cell migration and adhesion in ovarian cancer and as a secreted protein, ITGBL1 might be a novel biomarker for ovarian cancer diagnosis.     

急性髓系白血病转换为急性淋巴细胞白血病1例并文献复习

目的 探讨急性髓系白血病(acute myeloid leukemia, AML)转换为急性淋巴细胞白血病(acute lymphoblastic leukemia, ALL)的临床特征,为该病的临床诊断及治疗提供依据。方法 回顾性分析1例AML-M5b转换为B-ALL患者的临床资料、诊疗经过并复习相关文献。 结果 患儿,女,4岁。发热伴腹痛5天。结合患者病史、体格检查及辅助检查,诊断为AML-M5b,诱导化疗后达到血液学及细胞遗传学的完全缓解(complete remission, CR)。1年后患儿白血病复发,形态学及免疫表型符合B-ALL。通过嵌合抗原受体T细胞(chimeric antigen receptor expressing T cells, CAR-T)治疗,患儿再次获得了 CR,但不久后疾病复发,二次CAR-T治疗无效,患儿死亡。结论 急性白血病复发后发生系别转换,预后较差,需要根据复发后表型作相应的治疗调整,若AML转换为B-ALL,表达CD19,CAR-T治疗可使其再次获得CR,但易二次复发。临床上,对于复发性白血病,需要完善白血病免疫分型,基因突变检测,肿瘤染色体等细胞遗传性及分子生物学等方面的检查,以便更好地指导治疗及评估预后。     

儿童与成人急性淋巴细胞白血病临床特征的对比观察

目的 分析最近两年来儿童急性淋巴细胞白血病（ＡＬＬ）的临床预后并探讨其免疫学,细胞遗传学特征以及原发性多药耐药的发生率。方法 总结１５４例初治儿童ＡＬＬ的完全缓解（ＣＲ）率,同时动用免疫组织化学、细胞遗传学高等方法对白血病细胞的生物学特征进行研究。结果 儿童初治ＡＬＬ的平均ＣＲ率是９４．１％,显著高于成人（６７．８％）;免疫分型中,同时伴髓系表达者占１８．５％,Ｐｈ染色体的阳性率是４．８％,两组频     

PP242对Ph+急性淋巴细胞白血病细胞增殖的抑制作用及对AKT1-Ser473磷酸化的影响

目的观察ATP竞争性哺乳动物雷帕霉素靶蛋白复合物2(mTORC2)抑制剂PP242对体外培养的Ph染色体阳性(Ph+)急性淋巴细胞白血病(ALL)细胞增殖和AKT1473位丝氨酸磷酸化的影响。方法对Ph+ALL细胞株SUP-B15进行培养及传代,Cell Counting Kit-8比色法检测PP242(100 nM,250 nM,500 nM,750 nM,1 000 nM)处理细胞株SUP-B15后12 h、24 h、48 h细胞生长情况;应用免疫印迹法及实时定量RT-PCR检测使用PP242(100 nM,250nM,500 nM)培养细胞48 h后AKT1及磷酸化水平情况。结果 100 nM、250 nM、500 nM、750 nM、1 000 nM PP242对Ph+ALL细胞株SUP-B15作用12 h、24 h及48 h后,其抑制率:12 h组:10.17%,13.25%,20.13%,22.54%,27.61%;24h组:11.33%,20.54%,36.18%,41.66%,47.27%;48 h组:23.13%,39.24%,55.72%,61.88%,66.93%,与对照组相比差异均有统计学意义(P<0.05);100 nM、250 nM、500 nM PP242处理SUP-B15细胞48 h后,免疫印迹法检测结果显示AKT1蛋白的表达水平无明显变化,而473位丝氨酸(Ser473)磷酸化水平逐渐降低;实时定量RT-PCR检测结果显示,AKT1的mRNA表达无明显变化,绝对定量mRNA结果分别为:0.673±0.124,0.751±0.133,0.689±0.154,与正常对照组(0.679±0.167)相比差异无统计学意义(P>0.05)。结论 ATP竞争性mTORC2抑制剂PP242对体外培养的Ph+ALL细胞株SUP-B15细胞的增殖存在抑制作用,其作用强度呈浓度和时间依赖性;SUP-B15细胞中存在AKT1的表达;PP242在抑制Ph+ALL细胞株SUP-B15细胞的增殖过程中可下调AKT1Ser473磷酸化水平。     

骨髓间充质干细胞成骨分化中的微小RNA及Wnt通路

背景：有差异性表达的微小RNA在成骨细胞的增殖和分化中发挥广泛而又重要的作用,但其作用机制、作用靶点还不甚清楚,是近年来的研究热点。目的：对微小RNA通过Wnt信号通路调节成骨分化方面的研究现状及新进展作一综述。方法：应用计算机检索2000年1月至2012年1月CNKI和PubMed数据库,在标题和摘要中以＂MicroRNA,Wnt通路,成骨分化＂或＂MicroRNA,Wnt pathway,osteogenesis＂为检索词进行检索。选择文章内容与微小RNA经典Wnt通路促进成骨分化有关者,同一领域文献则选择近期发表或发表在权威杂志的文章。初检得到205篇文献,最终选择33篇文献进行综述。结果与结论：微小RNA是近年来生命科学的研究热点,越来越多的研究表明微小RNA作用于经典Wnt信号通路在成骨分化过程中发挥了重要作用。了解微小RNA在成骨分化中的调控机制是骨组织工程化培养的基础,具有巨大的应用前景。目前关于微小RNA的研究工作还处于起步阶段。     

NEP1-40对缺氧缺血性脑病新生大鼠的Wnt信号通路胞增殖的调控作用

目的探讨Nogo-A受体拮抗剂NEP1-40对Wnt信号通路和神经细胞增殖的调控作用。方法 40只大鼠被均分为HIBD(缺氧缺血性脑损伤)组和HIBD+NEP1-40组,采用PCR定量、Western blot分析、细胞增殖的免疫组化试验、8-异前列腺素评估等检测分析缺氧缺血性脑病新生大鼠的修复过程中Wnt信号通路中NgR的转录因子调控与神经细胞增殖。结果NEP1-40处理后,c Jun和c-Myc的表达在蛋白水平上调,基因表达水平上调,Ki-67增加,8-异前列腺素无显著变化。结论通过抑制NgR后发现,c-Jun和c-Myc是Wnt通路的主要转录因子,同时脑室下区神经细胞的增殖增加。     

姜黄素对多发性骨髓瘤细胞株MOLP-2/R的耐药逆转作用及与FA/BRCA途径的关系

目的 研究姜黄素与马法兰联用对人多发性骨髓瘤耐药细胞株MOLP-2/R的耐药逆转作用及其与FA/BRCA途径可能存在的关系,进一步探讨其作用机制.方法 采用MTT法检测细胞增殖抑制率;采用Western blot方法 检测FA/BRCA途径中的关键蛋白FANCD2单泛素化表达;采用流式细胞术测定细胞周期、细胞内药物浓度及细胞凋亡率.结果 姜黄素可增强马法兰对多发性骨髓瘤耐药细胞株MOLP-2/R的毒性,马法兰的IC50值由44.5 μmoL/L降至19.0 μmol/L.该作用是通过抑制FA/BRCA途径中的关键蛋白FANCD2单泛素化来实现的.姜黄素和马法兰联合作用组与单用马法兰组相比,MOLP-2/R细胞的凋亡率由(23.3±0.6)%增至(52.6±0.8)%,G2/M期细胞由9.1%增至18.5%,细胞内药物荧光强度由15.2±0.3增至21.4±0.8.而姜黄素单药作用则无此效应,也不伴有FA/BRCA途径受抑制.结论 姜黄素与小剂量马法兰合用可产生协同作用,姜黄素通过抑制FA/BRCA途径中的FANCD2单泛素化来逆转MOLP-2/R细胞对马法兰的耐药性.姜黄素介导的FA/BRCA途径受抑制,能增强马法兰对MOLP-2/R细胞的凋亡诱导及G2/M期阻滞作用,提高细胞内药物浓度.     

急性淋巴细胞白血病患儿亚甲基四氢叶酸还原酶基因多态性与甲氨蝶呤动态血药浓度相关性的研究

目的本研究旨在探讨亚甲基四氢叶酸还原酶(MTHFR)C677T、A1298C基因多态性对急性淋巴细胞白血病(ALL)患儿使用大剂量甲氨蝶呤(HD-MTX)化疗期间的MTX动态血药浓度的相关关系。方法 35例ALL患儿外周血,提取基因DNA,应用PCR-RFLP方法检测MTHFR C677T、A1298C基因型;应用荧光偏振免疫分析法(FPIA)24h、48h、72h监测患儿外周血中甲氨蝶呤动态血药浓度。结果 MTHFR C677T各基因型间24 h MTX浓度有差异(P0.05),携带CT型者明显高于携带CC型和TT型者;MTHFR C677T各基因型48 h、72 h的MTX浓度未见差异。MTHFR A1298C各基因型24 h、48 h7、2 h的MTX浓度未见差异(P0.05)。结论 MTHFR C677T基因多态性影响ALL患儿HD-MTX化疗期间MTX血药浓度,提示在HD-MTX治疗时可根据检测MTHFR C677T基因型进行个体化治疗。     

姜黄素抑制喉癌Hep2细胞系增殖的Rho/ROCK机制研究

目的分析姜黄素抑制喉癌Hep2细胞系增殖的相关机制及Rho/ROCK的参与作用。方法喉癌Hep2细胞随机分为4组:对照组、Y27632组、姜黄素低剂量组和高剂量组。姜黄素低剂量和高剂量组分别给予姜黄素(5μmol/L和50μmol/L)孵育,Y27632组在给予姜黄素孵育(10μmol/L)时给予Y27632(10μmol/L)。分析各组细胞活力情况,Western Blotting法分析各组细胞中细胞凋亡蛋白Bax、Bcl2、P53以及Caspase3/9及ROCK通路调控蛋白的表达变化。结果姜黄素可以明显抑制喉癌Hep2细胞的增殖,高剂量的姜黄素抑制率最高,同时,姜黄素高剂量可以显著增强Hep2细胞的Rho A、ROCK1/2及细胞凋亡相关蛋白的表达(P0.01),效果明显优于姜黄素低剂量组,Y27632可以明显恢复异常表达的上述蛋白。结论姜黄素通过激活ROCK信号通路和细胞凋亡蛋白上调表达抑制喉癌Hep2细胞的增殖,发挥肿瘤抑制的作用。     

A copper chelate induces apoptosis and overcomes multidrug resistance in T-cell acute lymphoblastic leukemia through redox imbalance and inhibition of EGFR/PI3K/Akt expression

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive form of cancer and the therapeutic outcome for T-ALL patients remains poor. Thus innovative therapeutic strategies with less toxic drugs are of immense need. Moreover combinational effect of redox imbalance with modulated EGFR/PI3K/Akt axis in T-ALL is still elusive. To explore more effective drugs we developed and characterized 5-SMAG, Cu-5-SMAG and Cu-OBPHA complexes by different spectroscopic methods and revealed that introduction of methoxy group and copper to the previously synthesized Schiff base ligand, NG can efficiently target leukemia by sparing the normal cells and overcomes MDR in T-ALL through induction of caspase3 dependent apoptosis as assessed by MTT, Cell-cycle, Annexin-V and caspase3 activation assay. However the ligand 5-SMAG fails to exert significant cytotoxicity. Moreover introduction of copper does not increase the efficacy of the drug molecule as Cu-OBPHA fails to exert significant effect compared to Cu-5-SMAG. Moreover Cu-5-SMAG targets T-ALL cells more than Cu-OBPHA because Cu-5-SMAG generates greater extent of redox imbalance compared to Cu-OBPHA and when this redox imbalance is reduced by application of NAC and PEG-Catalase, highest abrogation of apoptosis is observed following Cu-5-SMAG treatment In addition, Cu-5-SMAG significantly down-regulates the activation and expression of EGFR1, Akt and PI3K in drug-resistant T-ALL cells. Furthermore Cu-5-SMAG significantly increases the life-span of doxorubicin resistant and sensitive Ehrlich ascites carcinoma bearing Swiss albino mice without inducing any significant systemic toxicity compared to 5-SMAG and Cu-OBPHA treatment. Therefore typical architect of Cu-5-SMAG made it a promising new anti-leukemic agent irrespective of the MDR phenotype.     

肺耐药相关蛋白基因和多药耐药基因测定在急性淋巴细胞白血病中的意义

目的:评价肺耐药相关蛋白基因和多药耐药基因在急性淋巴细胞白血病患者中的表达情况与临床多药耐药的关系。方法:将47例急性淋巴细胞白血病患者分为临床非耐药组(A)和耐药组(B),用RT-PCR方法检测患者骨髓中肺耐药相关蛋白基因和多药耐药基因的mRNA表达。结果:肺耐药相关蛋白基因在A、B组阳性表达分别占6/27和8/20。统计学分析显示组间差异无显著性意义(χ2=1.15,P>0.05)。多药耐药基因在A、B组阳性表达分别占3/27和9/20。统计学分析显示组间差异有显著性意义(χ2=2.98,P<0.05)。在所有20例临床耐药患者中,5例同时表达上述两个基因。而临床非耐药组患者中,无一例同时表达上述两个基本基因。结论:肺耐药相关蛋白基因不能作为独立判定急性淋巴细胞白血病多药耐药的发生指标,多药耐药基因在判定急性淋巴细胞白血病多药耐药的发生中有一定临床意义。     

急性肺栓塞后线粒体前凋亡蛋白的表达及其促细胞凋亡机制研究

目的 研究大鼠急性肺栓塞(APE)后肺组织中线粒体前凋亡蛋白(ARTS)的表达变化及其诱导的细胞凋亡.方法 采用自体血栓栓塞颈总动脉制备大鼠APE模型.分别在APE前(对照)和APE后1、8、24和48 h开胸取肺脏.常规提取肺组织的总RNA和总蛋白,用半定量逆转录-聚合酶链反应(RT-PCR)方法检测ARTS mRNA表达水平,用蛋白质免疫印迹法(Western blotting)进一步验证ARTS以及ARTS诱导凋亡相关蛋白Bcl-2、Bcl-xL、XIAP和H2Ax的表达变化,用免疫组化法检测肺组织中ARTS在APE前后的表达变化及其组织分布情况,用原位末端缺刻标记法(TUNEL)检测肺组织内细胞凋亡情况.结果 APE后ARTS的mRNA及蛋白表达水平均升高,于1 h和48 h升高最明显(P相似文献   

Activation of PI3K is indispensable for interleukin 7-mediated viability, proliferation, glucose use, and growth of T cell acute lymphoblastic leukemia cells

Interleukin (IL)-7 is essential for normal T cell development. Previously, we have shown that IL-7 increases viability and proliferation of T cell acute lymphoblastic leukemia (T-ALL) cells by up-regulating Bcl-2 and down-regulating the cyclin-dependent kinase inhibitor p27kip1. Here, we examined the signaling pathways via which IL-7 mediates these effects. We investigated mitogen-activated protein kinase (MEK)-extracellular signal-regulated kinase (Erk) and phosphatidylinositol-3-kinase (PI3K)-Akt (protein kinase B) pathways, which have active roles in T cell expansion and have been implicated in tumorigenesis. IL-7 induced activation of the MEK-Erk pathway in T-ALL cells; however, inhibition of the MEK-Erk pathway by the use of the cell-permeable inhibitor PD98059, did not affect IL-7-mediated viability or cell cycle progression of leukemic cells. IL-7 induced PI3K-dependent phosphorylation of Akt and its downstream targets GSK-3, FOXO1, and FOXO3a. PI3K activation was mandatory for IL-7-mediated Bcl-2 up-regulation, p27kip1 down-regulation, Rb hyperphosphorylation, and consequent viability and cell cycle progression of T-ALL cells. PI3K signaling was also required for cell size increase, up-regulation of CD71, expression of the glucose transporter Glut1, uptake of glucose, and maintenance of mitochondrial integrity. Our results implicate PI3K as a major effector of IL-7-induced viability, metabolic activation, growth and proliferation of T-ALL cells, and suggest that PI3K and its downstream effectors may represent molecular targets for therapeutic intervention in T-ALL.