SDF-1、CXCR4和hs-CRP在儿童急性白血病中的表达

目的：检测儿童急性白血病骨髓中CXCR4、血清中SDF-1和hs-CRP的表达情况,探讨其临床意义,以积累临床经验,指导临床工作。方法：选取我院诊治的85例儿童急性白血病作为实验组,留取治疗前的骨髓穿刺标本,应用流式细胞术对其进行CXCR4的检测;留取治疗前的血清,对其进行SDF-1和hs-CRP的表达量的检测。选取同期20例非恶性血液病儿童的骨髓及血清作为对照组,检测骨髓中CXCR4和血清中SDF-1、hs-CRP的含量。比较二组中CXCR4、SDF-1和hs-CRP的表达及不同临床特征的意义。 结果：实验组CXCR4、SDF-1和hs—CRP的表达量显著高于对照组,CXCR4、SDF-1和hs—CRP在急性淋巴细胞性白血病患儿和髓外侵润的患儿表达明显增高。在实验组中CXCR4和SDF-1的表达具有正相关性。CXCR4和hs—CRP、SDF-1和hs—CRP的表达未见明显相关性。结论：CXCRd、SDF-1和hs—CRP在白血病患儿血清中高表达,其在次宿发生发展中可能具有重要意义,CXCR4和SDF-1在白血病的发生发展中具有协同作用。     

SDF-1/CXCR4轴在白血病患者骨髓中的表达及与血管新生的关系

目的：探讨SDF-1/CXCR4轴在白血病患者骨髓中的表达及与血管新生的关系。方法:采用 CD45/SSC设门四色流式细胞仪检测55例白血病患者骨髓的CXCR4表达。采用ELISA法检测SDF-1/VEGF;采用Envinson二步法检测骨髓组织MVD;多重PCR法检测白血病患者IgH、TCRγ链V区和J区,采用G-显带技术进行细胞遗传学核型分析。结果:AML组SDF-1(2145.21±329.45)pg/ml、CXCR4阳性表达率 (60.01±18.5)％;ALL组SDF-1(2549.02±303.4）pg/ml、 CXCR4阳性表达率（70.22±12.73）％;CML组SDF-1(1929.72±253.81)pg/ml、CXCR4阳性表达率（40.05±16.69）％;CRAL组SDF-1(2070.98±159.98)pg/mL、CXCR4阳性表达率(58.4±11.8)％;与对照组相比均明显增高,P<0.05。急淋SDF-1/CXCR4表达率高于其他白血病组;白血病SDF-1/CXCR4表达与相关因素的分析中发现,ALL与外周血WBC数量有相关性(r=0.534、 0.567, P<0.01),与急性白血病伴髓外浸润者有意义(P<0.05）;SDF-1/CXCR4 表达与白血病患者血小板计数、年龄、性别差异、骨髓增生程度、染色体改变、急性白血病骨髓细胞CD34+表达、ALL的IgH和TCRγ链V区和J区基因重排等因素无明显关系。白血病患者骨髓SDF-1、CXCR4、VEGF的相关系数为0.552, 0.553, 0.531,P<0.05。三者具有显著相关性。结论:白血病骨髓液中SDF-1含量及各群细胞CXCR4高表达,其中ALL表达最高;急性白血病缓解后依然高表达;SDF-1/CXCR4表达与ALL的外周血白细胞数量、有髓外浸润者有关,白血病中SDF-1/CXCR4、VEGF高表达并且相互之间存在有相关性,与白血病血管新生有关联。     

SDF-1等与非小细胞肺癌淋巴结转移相关性的探讨

目的:检测非小细胞肺癌(NSCLC)组织中SDF-1、CXCR4、VEGF-C和MMP-9的表达,探讨其及与淋巴结转移的关系.方法:采用免疫组化SP方法检测128例NSCLC组织中SDF-1、CXCR4、VEGF-C和MMP-9的表达,并分析其相关性.结果:SDF-1表达与病理分型(P＜0.05)及淋巴结转移(P＜0.01)相关,而与分化程度无关,P＞0.05. CXCR4表达与病理分型、淋巴结转移和分化程度皆相关,P＜0.05.VEGF-C表达与病理分型无关(P＞0.05),与淋巴结转移和分化程度相关,P＜0.05.MMP-9在肺癌中表达仅与淋巴结转移相关(P＜0.05),而与病理分型和分化程度无关,P＞0.05.相关性分析显示,SDF-1和CXCR4与VEGF-C高度相关(r值分别为0.704和0.751),提示SDF-1与VEGF-C之间存在相互调控作用.SDF-1与MMP-9相关,r=0.521.结论:SDF-1、CXCR4和VEGF-C协同调节,上调MMP-9表达,促进非小细胞肺癌淋巴结转移.     

骨髓增生异常综合征中CD34+细胞CXCR4的表达及其与细胞迁移的相关性

  目的  探讨不同危险度骨髓增生异常综合征（myelodysplasticsymdromes,MDS）中骨髓CD34+细胞CXCR4的表达情况及其与细胞迁移率的相关性。   方法  收集40例骨髓增生异常综合征患者的骨髓标本,根据IPSS积分系统进行危险度分组。低危组20例:IPSS积分0~1.5分;高危组20例:IPSS积分≥1.5分;同时采集10例健康者的骨髓标本作为对照。分离纯化骨髓CD34+细胞,通过流式细胞术检测CXCR4膜蛋白的表达;研究SDF-1α趋化作用下CD34+细胞的迁移率及CD34+细胞对骨髓基质细胞的迁移率。   结果  高危组MDS患者CD34+细胞CXCR4的表达率明显高于低危组和正常对照组（P < 0.000 1）;低危组和正常对照组之间CXCR4的表达率无显著性差异（P>0.05）。高危组CD34+细胞对SDF-1α及骨髓基质细胞的迁移率显著高于低危组及正常组（均P < 0.000 1）,且其对骨髓基质细胞的迁移率与CXCR4的表达呈正相关（P=0.000 1）。   结论  高危组MDS患者CD34+细胞CXCR4的表达量及其对骨髓基质细胞的迁移率均明显高于低危组患者,且其迁移率随CXCR4表达量的增加而升高,不同风险组的MDS患者存在SDF-1及其受体CXCR4表达和功能上的差异,SDF-1及其受体CXCR4在MDS发病中具有重要作用。      

急性白血病患者血清及脑脊液中基质细胞衍生因子-1α的含量及其意义

目的 检测急性白血病患者血清及脑脊液中的SDF-1α含量,分析其在不同分型的急性白血病中的变化及其临床意义.方法 选择急性白血病患者作为实验组,同期以颅脑外伤患者作为对照组.实验组采用标准方案化疗2周期.采集2组患者治疗前后的静脉血及脑脊液标本,采用酶联免疫吸附法(ELISA)检测入组患者血清及脑脊液中的SDF-1α含量.结果 治疗2周期后,实验组患者血清及脑脊液SDF-1α含量均低于治疗前(P＜0.05).治疗前后实验组与对照组比较,血清及脑脊液SDF-1α含量均明显升高(P＜0.05).与AML组比较,ALL组在治疗前后的血清及脑脊液SDF-1α含量较高;与非CNSL患者比较,CNSL组SDF-1α含量升高(P＜0.05).治疗2周期后,疗效为CR的患者SDF-1α含量较NR患者明显降低(P＜0.05).结论 SDF-1α与急性白血病的中枢神经系统浸润有关,在血清及脑脊液中含量变化可作为病情严重程度评估及预后判断的指标.     

趋化因子受体4和间质细胞衍生因子1α与鼻咽癌器官特异性转移的相关性研究

目的 研究趋化因子受体4(CXCR4)在鼻咽癌细胞中的表达,间质细胞衍牛因子1α(SDF-1α)在鼻咽癌远处靶器官中的表达,探讨CXCR4和(或)SDF-1α在鼻咽癌器官特异性转移中的作用.方法 应用逆转录聚合酶链反应(RT-PCR)和免疫组织化学法分析30例鼻咽癌、15例正常鼻咽组织中CXCR4 mRNA和蛋白的表达及其同临床病理学因素之间的相关性,应用免疫组织化学法分析鼻咽癌患者的正常颈部淋巴结(包括颈深上和颈深下淋巴结)、骨髓、肺、肝脏和肾脏、结肠(各5例)中SDF-1α蛋白的表达.结果 RT-PCR检测结果显示,鼻咽癌组织中CXCR4 mRNA相对表达强度(0.71±0.22)显著高于正常鼻咽组织(0.14±0.07;F=27.94,P<0.05);免疫组织化学检测结果显示,鼻咽癌组织中CXCR4蛋白的表达(1.58±0.59)显著高于正常鼻咽组织(0.51±0.22;F=17.75,P<0.05).鼻咽癌组织中CXCR4 mRNA和蛋白的表达与临床分期、淋巴结转移、细胞分化程度显著相关(均P<0.05).SDF-1α蛋白在鼻咽癌患者的颈深上淋巴结、骨髓、肺、肝脏中表达较高(2.35±0.67),而在颈深下淋巴结、肾脏和结肠中表达较弱(0.68±0.23),差异有统计学意义(t=10.13,P<0.01).结论 CXCR4的表达与鼻咽癌的转移密切相关,CXCR4和(或)SDF-1α在鼻咽癌器官特异性转移中可能发挥重要作用.     

血清降钙素原(PCT)联合PEWS预警用于急性白血病患儿脓毒症诊断的效能分析

目的:研究血清降钙素原(PCT)联合儿童早期预警评分（PEWS）用于急性白血病患儿脓毒症诊断的效能。方法:选取2017年07月至2020年08月于本院收治的100例急性白血病患儿作为研究对象，根据感染情况分为脓毒症组58例和非脓毒症组42例，采用酶联免疫法检测两组患儿的降钙素原（PCT）、超敏C反应蛋白（Hs-CRP）水平，比较两组儿童的降钙素原(PCT)、hs-CRP水平以及PEWS评分，利用ROC曲线分析血清PCT、hs-CRP水平、PEWS评分对急性白血病患儿脓毒症的诊断效能。结果:脓毒症组的血清PCT、hs-CRP水平、PEWS评分均显著高于非脓毒症组（P＜0.05）；重度组的血清PCT、hs-CRP水平、PEWS评分均显著高于轻度组（P＜0.05）；血清PCT、hs-CRP水平、PEWS评分联合检测诊断急性白血病脓毒症的AUC为0.924（95%CI:0.854~0.968），且联合诊断的AUC高于单一诊断（P＜0.05）；血清PCT、hs-CRP水平、PEWS评分联合检测鉴别轻、重度脓毒症的AUC为0.929（95%CI:0.830~0.980），且联合鉴别的AUC高于单一鉴别（P＜0.05）。结论:血清PCT、hs-CRP联合PEWS预警评分可有效诊断、鉴别急性白血病患儿脓毒症，有望成为脓毒症患儿的临床早期诊断和病情评估的指标。     

Stathmin蛋白的研究进展

目的: 探讨趋化因子受体CXCR4表达水平对人肺癌细胞转移潜能的影响.方法: 采用RT-PCR,FACS检测趋化因子受体CXCR4在肺癌细胞株95C,95D细胞的表达.构建CXCR4正义和反义真核表达质粒,用脂质体法转染至95C和95D细胞内,通过G418筛选出稳定转染株.通过趋化和趋化侵袭实验测定转染前后细胞对基质衍生因子(SDF-1α)的迁移能力,通过明胶酶谱法检测MMP-2活性,通过FACS检测细胞对血管内皮细胞的黏附能力.结果: CXCR4正义和反义真核表达质粒pcDNA3-X4和pcDNA3-ASX4能明显上调或下调肺癌细胞表面CXCR4的表达,上调95C细胞表面CXCR4的表达可以增强其对SDF-1α的趋化反应性、MMP-2活性及其对血管内皮细胞的黏附能力.相反,下调95D细胞表面CXCR4的表达抑制了其对SDF-1α的趋化反应性、MMP-2活性及其对血管内皮细胞的黏附能力.结论: 上调或下调肺癌细胞表面CXCR4表达可以显著增强或抑制其转移潜能,提示CXCR4表达水平与肺癌细胞转移潜能有关.     

CXCR4/SDF-1α对宫颈癌HeLa细胞定向迁移及增殖的影响

 目的了解CXCR4在HeLa细胞的表达情况,并借助细胞培养评价CXCR4/SDF-1α对HeLa细胞定向迁移及增殖的影响。方法CXCR4 mAb免疫染色HeLa细胞。用Transwell侵袭转移模型评价HeLa细胞的迁移情况,其中,上室中加入预先用(或不用)CXCR4单抗预孵育的HeLa细胞,下室中加入含0~100ng/mlSDF-1α的培养基。为评价CXCR4、SDF-1α对HeLa细胞增殖的影响,将HeLa细胞接种于有(无)SDF-1α和(或)CXCR4的低血清环境72h。结果CXCR4在所有HeLa细胞上均有表达。HeLa细胞能顺SDF-1α浓度差定向迁移,且这一作用可被CXCR4 mAb拮抗。SDF-1α能促进He-La细胞在低血清环境中增殖。结论CXCR4/SDF-1α参与了HeLa细胞定向迁移的过程并影响其增殖。     

基质细胞衍生因子-1及其受体CXCR4在白血病中表达的临床意义

 基质细胞衍生因子-1（SDF-1）是早期细胞生长因子,属趋化因子亚家族成员之一。其受体是CXCR4、SDF-1、CXCR4配体受体系统在血细胞的生成、干细胞归巢、血管新生以及白血病细胞的浸润等生理和病理过程中发挥重要作用。研究CXCR4在白血病中的表达及其与白血病浸润的关系,对丰富白血病形态学、免疫学、细胞遗传学（MIC）诊断分型的免疫学指标,为防治白血病浸润、复发而采取相关分子靶向治疗都具有重要意义。     

非小细胞肺癌中SDF-1、CXCR4的表达及临床意义

目的探讨基质细胞衍生因子(SDF1)及其受体CXCR4在非小细胞肺癌(NSCLC)中的表达及临床意义。方法利用免疫组织化学方法在40例NSCLC和10例对照组肺组织中检测SDF1、CXCR4的表达。结果在NSCLC中,SDF1、CXCR4的阳性表达率分别为65.00%(26/40)、45.00%(18/40),与对照组肺组织比较均有显著性差异(P<0.01,P<0.05)。SDF1、CXCR4的表达与肺癌细胞分化程度、淋巴结转移和组织学类型密切相关(P<0.01,P<0.05),而与性别、年龄无相关性(P>0.05),在NSCLC中,SDF1和CXCR4表达之间无显著相关性(P>0.05)。结论SDF1、CXCR4的表达与肺癌分化程度及转移有关,可作为临床预测肺癌患者预后的指标。     

Can inhibition of the SDF-1/CXCR4 axis eradicate acute leukemia?

Poor prognosis of acute leukemia with current treatments is mainly due to the relapse of the disease following chemotherapy. In the last decade, an emerging concept has proposed that the leukemia stem cells (LSCs) and their interactions with the BM microenvironment are the major cause of the acute leukemia relapse. Adhesion to the stromal niche is crucial for LSCs as it directly supports self-renewal, proliferation, arrest of differentiation and protects from damaging chemo-agents. One of the key players in this crosstalk between leukemic cells and the BM stroma niche is the chemokine SDF-1. SDF-1 regulates the process of homing and engraftment of LSCs into the BM and inhibition of its receptor CXCR4 induces leukemic cell mobilization into the circulation. However, besides its chemotactic and adhesive functions, SDF-1 is also a pleiotropic cytokine that regulates leukemic cell proliferation as well as their program of differentiation. CXCR4 antagonists are used in combination with chemotherapy in preclinical and clinical studies, which demonstrate that blocking CXCR4 is a novel promising approach of therapy. In this review, we focus on the multifaceted SDF-1/CXCR4 axis in acute leukemia and discuss how targeting this pathway could provide potential interest to eradicate the LSCs.     

Association of SDF-1 Gene Polymorphism with Increased Risk of Acute Myeloid Leukemia Patients

Background: Acute myeloid leukemia (AML) is a heterogenous group of disorders that emerge from the malignant transformation of hematopoietic stem cells. Chemokine stromal cell-derived factor 1(SDF-1) and its receptor CXC receptor 4 (CXCR4) has an essential role in dissemination of blast cells. Study aimed to detect CXCR4 expression and the SDF-1 (rs1801157) gene polymorphisms and correlate them with prognosis and outcome in AML patients. Subjects and Methods: The study was conducted on 60 de-novo AML patients, and 60 healthy controls. SDF-1 (rs1801157) gene polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and CXCR4 expression was done using flow cytometry analysis. Results: SDF-1 dominant model (AG+AA) had higher risk AML (p 0.002). CXCR4positive cases were associated significantly with toxic manifestations (p 0.019), lower CR rates (p 0.004), and unfavorable cytogenetics (p 0.027). Multivariate analysis showed that combined CXCR4positive with dominant SDF-1 considered as independent prognostic factor for shorter overall survival (OS) in AML patients (p 0.031). Conclusion: SDF-1 dominant model had a higher risk to develop AML, and CXCR4 positive expression predicts poor prognosis in AML patients and it could represent a targeted therapy in AML. In addition, CXCR4 could be easily integrated into the initial routine diagnostic work up of AML.     

CXCR4 regulates migration and development of human acute myelogenous leukemia stem cells in transplanted NOD/SCID mice

The chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 participate in the retention of normal hematopoietic stem cells within the bone marrow (BM) and their release into the circulation. Homing and engraftment of human stem cells in immunodeficient mice are dependent on cell surface CXCR4 expression and the production of BM SDF-1, which acts also as a survival factor for both human and murine stem cells. However, the role of SDF-1/CXCR4 interactions in the control of human acute myelogenous leukemia (AML) cell trafficking and disease progression is poorly understood. In this study, we report that although some AML cells do not express surface CXCR4, all AML cells tested express internal CXCR4 and SDF-1. Culture of AML cells with SDF-1 promoted their survival, whereas addition of neutralizing CXCR4 antibodies, SDF-1 antibodies, or AMD3100 significantly decreased it. Pretreatment of primary human AML cells with neutralizing CXCR4 antibodies blocked their homing into the BM and spleen of transplanted NOD/SCID/B2m(null) mice. Furthermore, weekly administrations of antihuman CXCR4 to mice previously engrafted with primary AML cells led to a dramatic decrease in the levels of human AML cells in the BM, blood, and spleen in a dose- and time-dependent manner. Interestingly, the same treatment did not affect significantly the levels of normal human progenitors engrafted into NOD/SCID mice. Taken together, our findings demonstrated the importance of the SDF-1/CXCR4 axis in the regulation of in vivo motility and development of human AML stem cells and identified CXCR4 neutralization as a potential treatment for AML.     

Functional CXCR4-expressing microparticles and SDF-1 correlate with circulating acute myelogenous leukemia cells

Stromal cell-derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 are implicated in the pathogenesis and prognosis of acute myelogenous leukemia (AML). Cellular microparticles, submicron vesicles shed from the plasma membrane of various cells, are also associated with human pathology. In the present study, we investigated the putative relationships between the SDF-1/CXCR4 axis and microparticles in AML. We detected CXCR4-expressing microparticles (CXCR4(+) microparticles) in the peripheral blood and bone marrow plasma samples of normal donors and newly diagnosed adult AML patients. In samples from AML patients, levels of CXCR4(+) microparticles and total SDF-1 were elevated compared with normal individuals. The majority of CXCR4(+) microparticles in AML patients were CD45(+), whereas in normal individuals, they were mostly CD41(+). Importantly, we found a strong correlation between the levels of CXCR4(+) microparticle and WBC count in the peripheral blood and bone marrow plasma obtained from the AML patients. Of interest, levels of functional, noncleaved SDF-1 were reduced in these patients compared with normal individuals and also strongly correlated with the WBC count. Furthermore, our data indicate NH(2)-terminal truncation of the CXCR4 molecule in the microparticles of AML patients. However, such microparticles were capable of transferring the CXCR4 molecule to AML-derived HL-60 cells, enhancing their migration to SDF-1 in vitro and increasing their homing to the bone marrow of irradiated NOD/SCID/beta2m(null) mice. The CXCR4 antagonist AMD3100 reduced these effects. Our findings suggest that functional CXCR4(+) microparticles and SDF-1 are involved in the progression of AML. We propose that their levels are potentially valuable as an additional diagnostic AML variable.     

POL5551, a novel and potent CXCR4 antagonist,enhances sensitivity to chemotherapy in pediatric ALL

The importance of the cell surface receptor CXCR4 and the chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) is well-established in normal and malignant hematopoiesis. The Protein Epitope Mimetic POL5551 is a novel and potent antagonist of CXCR4. POL5551 efficiently mobilizes hematopoietic stem and progenitor cells, but its effects in acute lymphoblastic leukemia (ALL) have not been reported. Here, we demonstrate that POL5551 is a potent antagonist of CXCR4 in pre-B and T cell ALL cell lines and pediatric ALL primary samples. POL5551 has activity at nanomolar concentrations in decreasing CXCR4 antibody binding, blocking SDF-1α-mediated phosphorylation of ERK1/2, inhibiting SDF-1α-induced chemotaxis, and reversing stromal-mediated protection from chemotherapy. POL5551 is significantly more effective at inhibiting CXCR4 antibody binding than the FDA-approved CXCR4 inhibitor plerixafor in ALL cell lines and primary samples. We also show that treatment with POL5551 in vitro and cytarabine +/− POL5551 in vivo modulates surface expression of adhesion molecules, findings that may guide the optimal clinical use of POL5551. Finally, we demonstrate that POL5551 increases sensitivity to cytarabine in a xenograft model of a high-risk pediatric ALL, infant MLL-rearranged (MLL-R) ALL. Therefore, disruption of the CXCR4/SDF-1 axis with POL5551 may improve outcomes in children with high-risk ALL.