自分泌运动因子及其受体在乳腺癌中的表达

目的了解自分泌运动因子(autocrine motilityfactor,AMF)及其受体(autocrine motilityfactor receptor,AMFR)在乳腺癌组织中的表达及其与乳腺癌临床病理特征之间的关系.方法应用免疫组化SABC方法研究25例正常乳腺组织、23例乳腺上皮增生组织、71例乳腺癌组织中AMF、AMFR的表达.结果AMF与AMFR阳性结果完全一致.正常乳腺组织、乳腺儋上皮增生组织AMF、AMFR阳性率均低于癌组织(P＜0.05).71例原发性乳腺癌组织中有45例(63%)阳性.浸润性导管癌AMF、AMFR水平明显高于导管内癌(P=0.001).AMF、AMFR高表达与乳腺癌组织学分级、TNM分期有关(P＜0.05),与年龄、肿块大小、淋巴结是否转移无关(P＞0.05).结论AMF、AMFR表达水平高与乳腺癌的进展有关,具有判断预后的价值.     

果糖二磷酸醛缩酶A在急性髓系白血病患者骨髓中的表达及其对预后的影响

目的探讨果糖二磷酸醛缩酶A(ALDOA)在急性髓系白血病(AML)患者骨髓中的表达及其与患者临床特征和预后之间的关系。方法收集2013年1月至2015年12月郑州大学第一附属医院及郑州大学附属儿童医院90例AML(非急性早幼粒细胞白血病)患者(AML组)及18例异基因造血干细胞移植供者(正常对照组)骨髓标本, 应用实时荧光定量聚合酶链反应(qRT-PCR)检测ALDOA mRNA相对表达量。对患者临床资料进行回顾性分析, 根据疗效与随访结果将患者分为持续完全缓解组和难治复发组。分析AML组与正常对照组及持续完全缓解组与难治复发组之间ALDOA mRNA相对表达量的差异。应用单因素及多因素Cox比例风险模型分析AML患者预后的影响因素。结果 AML组ALDOA mRNA相对表达量高于正常对照组(5.71±0.44比1.10±0.08), 差异有统计学意义(t=4.74, P<0.001);难治复发组ALDOA mRNA相对表达量高于持续完全缓解组(6.69±0.67比4.30±0.36), 差异有统计学意义(t=2.79, P<0.001)。不同白细胞计数、骨髓原始细胞比例及...     

Axl在急性髓系白血病中的表达及其临床意义

目的:通过检测Axl受体酪氨酸激酶在急性髓系白血病(AML)细胞中的表达水平,探讨Axl在AML多药耐药中的作用及其临床意义.方法:采用实时定量RT-PCR方法检测24例初治、11例复发/难治、12例完全缓解成人AML患者及11例对照者骨髓单个核细胞Axl mRNA的表达水平,并分析其在不同分组间的表达规律及与各临床特征、缓解率之间的关系.结果:初治组及复发/难治组Axl mRNA表达水平均高于对照组(Z 分别为-2.791、-2.310,P 均＜0.05);复发/难治组Axl mRNA表达水平明显高于初治组Axl mRNA表达水平(Z=-2.239,P=0.025);初治组Axl mRNA表达水平也显著高于完全缓解组(Z=-1.956,P=0.040);而对照组与完全缓解组之间比较差异无统计学意义(Z=-0.895,P=0.325).Axl的表达水平与初发未治AML患者性别、年龄、初诊白细胞数、骨髓原始细胞百分数、染色体核型及免疫表型无明显相关性(P 均＞0.05);初治组经2个疗程化疗后,完全缓解组(CR)骨髓单个核细胞Axl mRNA表达量高于未完全缓解组(PR+NR),差异有统计学意义(Z=-2.104,P=0.035).结论:Axl mRNA高表达与AML患者的多药耐药及病情进展密切相关.     

急性白血病患者骨髓MK和VEGF基因表达及临床意义的探讨

目的:研究急性白血病(AL)患者MK和VEGF基因的表达水平及其与血管新生、临床预后的关系.方法:应用实时荧光定量PCR检测95例不同病程的AL患者骨髓单个核细胞MK和VEGF mRNA的表达.结果:急性非淋巴细胞白血病(AML)和急性淋巴细胞白血病(ALL)患者MK和VEGF表达量均明显高于对照组,P=0.000 0.初治/复发组AL患者MK、VEGF基因表达水平较对照组及完全缓解组升高,完全缓解组MK和VEGF表达水平仍较对照组高,P均＜0.01.缓解组MK表达水平与VEGF和骨髓原幼细胞数存在显著相关性,r=0.673,P＜0.01; r=0.468,P＜0.01.复发组MK和VEGF之间也存在相关性,r=0.526,P＜0.05.初治组和对照组MK、VEGF、骨髓原幼细胞数之间无明显相关性.结论:MK、VEGF高表达可能与AL血管新生、病情进展有关,且两者存在一定的协同作用,共同监测可能对研究急性白血病的发病及指导治疗和估计预后有一定意义.     

XIAP和XAF1在急性白血病中的表达及其意义

目的:研究急性白血病(AL)患者中凋亡抑制蛋白XIAP和XIAP相关因子1(XAF1)的表达及其临床意义.方法: 以86例AL患者为研究对象,其中初诊未治患者56例,治疗后完全缓解期(CR)患者22例,缓解后复发患者8例,应用RT-PCR方法检测骨髓细胞中XIAP mRNA和XAF1mRNA表达.对照组为20名门诊非恶性血液病患者.结果: 对照组、初治组、缓解组和复发组的XIAP mRNA平均表达水平之间比较,差异具有显著性(F=58.77,P<0.01);对照组的XIAP mRNA平均表达水平为0.29±0.13,明显低于AL初治患者(0.98±0.30)(P<0.01)和缓解组(0.45±0.17)(P<0.05);缓解组的平均表达水平明显低于初治组(P<0.01);复发组的XIAP mRNA平均表达水平最高(1.23±0.30),不仅明显高于对照组和缓解组(P<0.01),也高于初治组(P<0.05). 对照组的XAF1 mRNA的阳性表达率为100%,明显高于初治组(51.79%)(P<0.01)和缓解组(77.27%)(P<0.05);初治组和复发组的阳性表达率(25%)明显低于缓解组(P<0.05),但初治组与复发组无显著性差异(P>0.05).XIAP mRNA和XAF1 mRNA表达在初治AML和ALL组比较均无显著性差异(P>0.05).结论: XIAP异常高表达和XAF1异常低表达可能在急性白血病发病和病情进展中起重要作用,为AL 的治疗提供了新思路.     

VEGF和MDR1在急性髓系白血病中的表达及其与预后关系研究

目的研究初治急性髓系白血病(AML)患者化疗前后血清VEGF的表达及骨髓白血病细胞MDR1的表达。探讨VEGF与MDR1的表达对AML预后中的评价作用。方法血清VEGF的含量采用双抗体夹心酶联免疫吸附法检测,MDR1的表达采用直接免疫荧光标记单抗流式细胞仪(FCM)检测。结果初发和未缓解AML患者血清VEGF水平明显高于正常对照组和AML完全缓解(CR)组(P<0.01);未缓解患者血清VEGF含量与CR组比较差异有显著性意义(P<0.01),而与初治组比较差异无显著性意义(P>0.05)。初发AML患者骨髓单个核细胞的P-gp表达为(9.20±3.13)%。初发和未缓解AML患者骨髓白血病细胞的P-gp表达高于正常对照组和AML缓解组(P<0.01);P-gp表达与VEGF表达具有一定的相关性(r=0.639,P<0.01)。结论AML患者血清VEGF水平一定程度上可能反映体内白血病细胞的总负荷量同时与临床病情变化密切相关,可作为判断预后的指标之一。AML患者P-gp阳性表达者疗效差、预后不良。AML患者VEGF与P-gp的表达呈正相关,同时检测这两项指标对于AML的预后判断较单独检测更有意义。     

急性白血病细胞肺耐药相关蛋白的表达及其临床意义

目的:研究急性白血病细胞肺耐药蛋白(lung resistance-related protein,LRP)的表达,观察其表达率与化疗缓解率的关系。方法:利用LRP单克隆抗体,采用流式细胞术(FCM)分别测定15位正常对照和79例急性白血病(a-cute leukemia,AL)患者LRP的表达率,并分析其表达的临床意义。结果:初治急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)组LRP的表达率为(29.29±6.72)%,高于初治急性髓细胞白血病(acute myelogenous leukemia,AML)组的(21.46±5.23)%,P=0.0182;复发/难治ALL组LRP的表达率为(41.18±9.06)%,也较复发/难治AML组的(30.44±11.51)%高,P=0.0154。复发/难治组LRP的表达率均高于相应的初治组,P值分别为0.0175和0.0138。急性白血病细胞LRP( )组的临床缓解率为30.8%,明显低于LRP(-)组的77.5%,P=0.0081。结论:复发/难治组LRP的表达率高于初治组,LRP过度表达与急性白血病患者的临床耐药及临床疗效密切相关。     

SHP-1与SOCS3基因在成人急性白血病中的表达及其与预后的关系

目的 了解SHP-1基因和SOCS3基因在白血病细胞中的表达及其对预后的影响,为进一步阐明SHP-1和SOCS3基因在白血病发病中的作用机制打下基础.方法 用实时荧光定量PCR(FQ-PCR)和RT-PCR方法分别对AL患者和13例正常对照的骨髓进行SHP-1、SOCS3 mRNA水平监测.结果 ①正常人SHP-1 mRNA均呈阳性表达,其表达水平为3.413±2.018,ALL患者SHP-1均为阴性表达;初治、复发AL组SHP-1表达水平明显低于正常对照组(P<0.01),缓解后SHP-1 mRNA水平上升但仍低于正常时照(P<0.01).②初治和复发AL患者SOCS3 mRNA表达水平均明显高于缓解组和正常对照组(P<0.05);缓解患者SOCS3 mRNA与正常对照相比无统计学意义(P>0.05).③SHP-1表达阳性组缓解率(76.4%)明显高于表达阴性组(47.3%)(P<0.01).结论 SHP-1和SOCS3均参与白血病发病,但发病机制不同.SHP-1与白血病的疗效预后有关.     

急性髓系白血病Evil基因表达的研究

目的:探讨急性髓系白血病中Evil基因表达及意义.方法:应用RT-PCR方法检测了94例急性髓系白血病(AML,包括初治19例,复发23例,缓解22例)患者及10名正常对照Evil基因的表达.结果:10名正常人骨髓单个核细胞中无Evil mRNA的表达.AML患者Evil基因总的阳性表达率为18.1%(17/94),M5型未见表达,其中初治、复发、缓解组的Evil基因阳性率分别为26.5%、17.4%、0,初治和复发组差别无显著性(P=0.554).M3患者中Evil、PML/RARα双阳性组与PML/RARα单阳性组相比复发率高(P＜0.05),早期死亡率高.Evil阳性的AML患者多数生存期短.结论:Evil基因是一个原癌基因,在髓系白血病的发病中起重要作用,是髓系白血病预后不良的一个指标.     

T细胞免疫球蛋白黏液素3基因在急性白血病中的表达及其临床意义

目的 探讨T细胞免疫球蛋白黏液素3(Tim-3)基因在急性白血病患者和白血病细胞株中的表达情况,并分析其临床意义.方法 收集初治急性白血病患者65例和8株白血病细胞株,同时收集15名健康供体的骨髓作为健康对照,分离单个核细胞,提取DNA,应用实时荧光定量聚合酶链反应检测单个核细胞上Tim-3 mRNA相对表达量,比较不同类型白血病Tim-3 mRNA表达差异,分析急性白血病患者Tim-3 mRNA表达与临床特征的关系.结果 M3细胞株NB4及非M3细胞株Kasumi-1、HL60、SHI-1、THP-1、Jurkat、CCRF-CEM、Mutz-1 Tim-3 mRNA相对表达量分别为(4.51±0.62)×10-5、(79.22±2.91)×10-5、(41.14±3.01)×10-5、(27.70±3.50)×10-5、(60.47±4.97)×10-5、(10.44±1.77)×10-5、(42.80±2.95)×10-5、(11.68±1.96)×10-5.非M3细胞株Tim-3 mRNA表达水平均高于NB4细胞株(均P＜0.05).非M3型急性髓系白血病(AML)患者Tim-3 mRNA相对表达量为(27.64±13.35)×10-4,AML-M3患者为(4.81±2.30)×10-4,急性淋巴细胞白血病(ALL)为(32.09±23.42)×10-4.其在非M3型AML和ALL中的表达水平高于健康对照[(12.02±4.22)×10-4],差异有统计学意义(P＜0.05),非M3型AML和ALL患者的Tim-3 mRNA表达水平差异无统计学意义(P＞0.05).在非M3型AML中,根据法、美、英协作组(FAB)分型,Tim-3 mRNA在AML-M4中的表达水平高于其他类型AML(P＜0.05).不同性别、年龄、骨髓原始细胞数、初诊白细胞数的急性白血病患者间Tim-3 mRNA表达水平差异均无统计学意义(均P＞0.05).结论 Tim-3 mRNA高表达于非M3型的急性白血病患者和细胞株,其表达水平与初治急性白血病患者的性别、年龄和原始细胞数无关.     

Autocrine motility factor and its receptor: Role in cell locomotion and metastasis

Summary The ability to locomote and migrate is fundamental to the acquisition of invasive and metastatic properties by tumor cells. Autocrine motility factor (AMF) is a 55 kD cytokine produced by various tumor cells which stimulates their in vitro motility and in vivo lung colonizing ability. AMF stimulates cell motility via a receptor-mediated signalling pathway. Signal transduction following binding of AMF to its receptor, a cell surface glycoprotein of 78 kD (gp78) homologous to p53, is mediated by a pertussis toxin sensitive G protein, inositol phosphate production and the phosphorylation of gp78. Cell surface gp78 is localized to the leading and trailing edges of motile cells but following cell permeabilization is found within an extended network of intracellular tubulovesicles. Gp78 tubulovesicles colocalize with microtubules and extension of the tubulovesicular network to the cell periphery is dependent on the presence of intact microtubules. Gp78 labeled vasicles can be induced to translocate between the cell center and periphery by altering intracellular pH as previously described for tubulovesicles labeled by fluid phase uptake. Anti-gp78 mAb added to viable motile cells is localized to large multivesicular bodies which, with time, relocate to the leading edge. Binding of AMF to its receptor induces signal transduction, similar to chemotactic stimulation of neutrophil mobility, as well as the internalization and transport of its receptor to the leading edge stimulating pseudopodial protrusion and cell motility.     

Receptors for platelet derived growth factor in human glioma cell lines and influence of suramin on cell proliferation

Summary A panel of 11 established human glioma cell lines was used to evaluate PDGF receptor binding using radio-idinated biosynthetic PDGF-AA and PDGF-AB as primary ligands. It was found that PDGF-receptor-binding was qualitatively heterogeneous. The affinities for PDGF-AA as well as PDGF-AB binding were within a close range of 0.13–0.33 nM and 0.16–1.1 nM, respectively. The number of binding sites per cell ranged between 56.000 and 250.000 for PDGF-AA and 72.000 to 300.000 for PDGF-AB. Two lines had only background levels of PDGF-AA binding. PDGF-AB binding was the dominant binding component in all but one cell line. In seven cell lines there were two binding components upon saturation analysis consisiting of a high affinity component and a non-saturable low affinity component.PDGF and PDGF-receptors are suspected to be part of an autocrine loop in gliomas. Therefore, the effect of suramin on cell proliferation in serumfree cultures was tested in the same cell lines using doses of 25, 200 or 500 g/ml. It was found that the response to suramin was variable and that two cell lines still reached 2.8 fold and 4.5 fold their initial cell density even in the presence of 500 g/ml whereas all other cells were completely arrested. Analyzing the response to 200 g/ml it became evident, that the PD GF binding characteristics are of no reliable predictive value in respect to the efficacy of suramin.     

肝癌细胞中血管内皮生长因子的自分泌及其意义

目的 研究人肝癌细胞中血管内皮生长因子(VEGF)的自分泌及其意义。方法：反转录PCR(RTPCR)检测体外培养的人肝细胞肝癌细胞系SMMC7721、HHCC和HepG2中VEGF受体的表达。采用人工合成的反义寡核苷酸(ODN)抑制肝癌细胞VEGF的表达后，分别检测肝癌细胞增殖活性和超微结构的变化，流式细胞仪分析肝癌细胞周期以及VEGF’受体蛋白表达的变化。结果：VEGF受体1(Flt-1)在SMMC7721细胞中有表达，而HHCC和HepG2细胞则表达VEGF受体2(KDR)。反义ODN对HHCC细胞的增殖有抑制作用，且与ODN的作用浓度呈线性关系，浓度为2．5μmol／L、5 μmol／L和10μmol/L时细胞增殖抑制率分别为26％、41％和50％。反义ODN对SMMC7721细胞则无此作用，而正义ODN对上述2种细胞均无作用。反义ODN作用后，HHCC细胞出现S期比率下降，并且在细胞周期曲线中出现凋亡前期峰，而SMMC7721细胞无上述变化。反义0DN能降低HHCC细胞膜表面KDR的表达(阳性率分别为：反义组19.2％，正义组29.8％，对照组31.2％)，对于SMMC7721细胞Flt-1的膜表面表达则无明显影响。结论：人肝癌细胞中存在VEGF的自分泌途径，其机制可能为通过诱导肝癌细胞膜表面KDR的表达及其信号转导作用，从而调节肝癌细胞的分裂增殖。     

The road less travelled: c-kit and stem cellfactor

Autocrine stimulation of growth factor receptors by autonomouslyproduced ligands regulates different aspects of cellular transformationand progression. In several tumors, including gliomas, multipleautocrine systems are activated and may exert differentfunctions in the malignant transformation process. The c-kitproto-oncogene is widely expressed in human gliomas, andit may be activated by its co-expressed ligand,stem cell factor (SCF). Studies in glioma celllines as well as different tumor types suggestthe possibility of intracellular interactions of c-kit withSCF. Although c-kit and SCF may not playa primary and causal role in the initiationand progression of glial tumors they may stillbe contributing factors in glioma biology. It canbe hypothesized that the parallel activation of severalautocrine systems including some of which have foundless attention in gliomas, such as c-kit/SCF, couldcompromise the efficacy of therapies targeting different autocrineloops. A better understanding of the multiplicity andmechanisms of autocrine stimulation has implications for thedevelopment of new therapies interfering with autocrine tumorcell growth.     

Transformation of an established mouse mammary epithelial cell line following transfection with a human transforming growth factor alpha cDNA

To determine whether the enhanced expression of transforming growth factor alpha (TGF alpha) is sufficient to induce the neoplastic transformation of an immortalized population of mammary epithelial cells, we cotransfected NOG-8 cells, a cloned mouse mammary epithelial cell line, with a simian virus 40-human TGF alpha cDNA expression vector plasmid and a pSV2neo plasmid. After cotransfection, nine G418-resistant NOG-8 colonies were cloned and expanded. All clones were subsequently analyzed for TGF alpha mRNA expression by northern blot analysis, TGF alpha secretion, anchorage-dependent growth in serum-free medium, anchorage-independent growth in soft agar, and tumorigenicity in nude mice. Three TGF alpha-transfected NOG-8 clones expressed high levels of a specific TGF alpha mRNA, secreted elevated levels of TGF alpha into the culture medium (177-595 ng/10(8) cells/48 h), exhibited an enhanced growth rate, grew aggressively as colonies in soft agar, and formed undifferentiated, invasive carcinomas in nude mice. A neutralizing mouse monoclonal antibody generated against the low molecular weight human TGF alpha peptide was able to inhibit colony formation in soft agar by TGF alpha-transfected NOG-8 clones that produced high levels by TGF alpha. This inhibition suggested that TGF alpha acted through an external autocrine loop. NOG-8 cells and NOG-8 cells transfected with a pSV2neo plasmid alone secreted very low levels of TGF alpha, failed to grow as colonies in soft agar and did not form tumors in nude mice. These results demonstrate that overexpression of a human TGF alpha cDNA in immortalized, nontransformed mouse mammary epithelial cells can induce a transformed phenotype in vitro and can facilitate tumor formation in vivo.     

Endogenous serum levels and surface receptor expression of GM-CSF and IL-3 in patients with myelodysplastic syndromes

The majority of patients suffering from myelodysplastic syndromes (MDS) die of complications due to cytopenia. Clinical trials have demonstrated that in an essential number of MDS patients cytopenia can be ameliorated by exogenously supplied growth factors. We investigated endogenous serum levels of GM-CSF and IL-3 in 15 healthy individuals and 34 patients suffering from MDS. No circulating growth factors were detected in the serum of healthy controls, nor was IL-3 measurable in MDS patients. GM-CSF serum levels, however, were elevated in a significant number of patients (26.5%). Levels did not correlate with FAB classification, leukocyte count or chromosomal abnormalities. No significant differences in GM-CSF or IL-3 receptor expression were detected between healthy individuals and MDS patients. One patient with increased endogenous GM-CSF serum level and normal surface receptor expression responded to exogenously applied GM-CSF. In the light of these results, a funetional alteration of growth factor receptors or disturbances of signal transduction pathways must be discussed for MDS.