PI3K/AKT信号通路在RANKL诱导的SGC-7901胃癌细胞迁移中的作用

目的文献报道RANKL/RANK/OPG途径与肿瘤细胞迁移及骨转移密切相关,但RANKL/RANK途径是否参与胃癌细胞迁移,尚无文献报道。本文拟检测RANK在胃癌细胞系SGC-7901细胞中的表达,并进一步探讨磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/Akt)信号通路在RANKL诱导的胃癌细胞迁移中的作用。方法 West-ern blot检测SGC-7901细胞表面RANK蛋白的表达;RANKL刺激后磷酸化Akt(P-Akt)及Akt的表达;Transwell法测定RANKL及抑制剂刺激后细胞迁移能力的改变。结果 SGC7901细胞表达RANK蛋白。RANKL(1μg/mL)诱导SGC-7901细胞迁移能力增强,迁移增加率为57.2%±5.9%,RANKL抑制剂rOPG(5μg/mL)显著抑制RANKL诱导的细胞迁移(13.88%±3.57%,P<0.05)。RANKL刺激后30 min～3 h,SGC-7901细胞p-Akt表达升高,应用PI3K的抑制剂LY294002(50 mmol/L)显著抑制RANKL诱导的胃癌细胞SGC-7901的迁移(57.28%±5.91%vs23.18%±2.79%,P<0.05)。结论胃癌细胞系SGC-7901细胞表达受体RANK,PI3K/Akt信号通路参与RANKL诱导的SGC-7901细胞迁移。     

Localization of CRMP5 mRNA by in situ hybridisation during development of the mouse forebrain

The expression of the collapse response mediator protein CRMP5 in the prenatal mouse is largely unknown. Evidence suggests that CRMP family members play important roles in neurite outgrowth, and CRMP5 is known to modulate outgrowth of processes in oligodendrocytes through signalling via neuropilin-1 and SemaA. Furthermore, CRMP family members function in axon regeneration after injury and are implicated in the early stages of Alzheimer's disease. Despite these findings relatively little is known about the specific roles these proteins play. The aim of the present study was to evaluate CRMP5 expression in the developing mouse forebrain using in situ hybridisation. Serial coronal sections of brain from E12.5 to E18.5 were analysed. We found highly specific patterns of expression which were restricted to the post-mitotic layers of both the ganglionic eminence and neocortex, and an additional domain of strong expression in the pyramidal layers of the hippocampus in all prenatal ages. Our results are therefore consistent with a role for CRMP5 in process extension. Interestingly, our results also revealed a temporal switch in high-expression levels from the ganglionic eminence to the cortex at a critical time during tangential cell migration. However, the pattern of expression appeared more representative of a general permissiveness for neurite outgrowth rather than one which is restricted to a particular cell subset or cell class. Additionally, expression was also found during periods predominated by neurogenesis and not neurite extension. We conclude that expression of CRMP5 is consistent with a dynamic implicit role in forebrain development.     

Wnt11r is required for cranial neural crest migration

wnt11r is a recently identified member of the Wnt family of genes, which has been proposed to be the true Xenopus homologue to the mammalian wnt11 gene. In this study we have examined the role of wnt11r on neural crest development. Expression analysis of wnt11r and comparison with the neural crest marker snail2 and the noncanonical Wnt, wnt11, shows wnt11r is expressed at the medial or neural plate side of the neural crest while wnt11 is expressed at the lateral or epidermal side. Injection of wnt11r morpholino leads to strong inhibition of neural crest migration with no effect on neural crest induction or maintenance. This effect can be rescued by co‐injection of Wnt11r but not by Wnt11 mRNA, demonstrating the specificity of the loss of function treatment. Finally, neural crest graft experiments show that wnt11r is required in a non–cell‐autonomous manner to control neural crest migration. Developmental Dynamics 237:3404–3409, 2008. © 2008 Wiley‐Liss, Inc.     

急性胰腺炎大鼠模型巨嗜细胞移动抑制因子(MIF) 表达变化规律的研究

目的 :本实验旨在观察急性胰腺炎大鼠MIF表达变化规律 ,探讨MIF在急性胰腺炎发病机制中的作用。方法 :使用雄性Wistar大鼠建立急性水肿性胰腺炎 (AEP)与急性坏死性胰腺炎 (ANP)动物模型。分别于Ohr,1hr,2hr,4hr,8hr采集AEP组与ANP组下腔静脉血 ,ANP组腹水标本。ELISA法测定各组标本MIF水平含量。同时检测AEP组与ANP组血清淀粉酶含量。结果 :与正常组相比 ,AEP组血液MIF含量升高 (P <0 0 5 ) ,ANP组血液与腹水MIF含量均升高 (P <0 0 1) ;与正常组相比 ,AEP组与ANP组血清淀粉酶明显升高 ,有统计学意义 (P <0 0 5 )。结论 :急性胰腺炎发病过程中MIF表达明显增加 ,提示在急性胰腺炎发病机制中MIF可能发挥了作用。     

巨噬细胞移动抑制因子和基质金属蛋白酶2在胶质瘤组织中的表达及其临床意义

目的探讨巨噬细胞移动抑制因子(MIF)和基质金属蛋白酶2(MMP-2)在胶质瘤组织中的表达及其临床意义。方法收集南华大学附属第一医院神经外科2009—2013年收集的经病理检查证实的胶质瘤石蜡标本64份作为胶质瘤组,另选取同期因脑创伤行颅内减压去除的脑组织标本36份作为对照组,采用免疫组化SP法检测两组MIF及MMP-2的表达情况,并分析胶质瘤组织中MIF与MMP-2表达的相关性。结果胶质瘤组MIF、MMP-2阳性表达率高于对照组(P0.01)。不同性别、年龄胶质瘤患者MIF、MMP-2阳性表达率比较,差异无统计学意义(P0.05),但胶质瘤组织学Ⅲ~Ⅳ级者MIF、MMP-2阳性表达率高于Ⅰ~Ⅱ级者(P0.05)。胶质瘤组织中MIF与MMP-2表达呈正相关(r=0.76,P0.05)。结论 MIF和MMP-2过表达可能在胶质瘤的发生发展中起重要作用,并与胶质瘤的恶性程度密切相关。     

MIF和iNOS在胎膜早破产妇胎膜组织中的表达及意义

目的探讨巨噬细胞移动抑制因子（MIF）与诱导型一氧化氮合酶（iNOS）在胎膜早破（PROM）发生、发展中的作用。方法研究对象为90例剖宫产的产妇;其中早产胎膜早破（p PROM）30例（p PROM组）,足月胎膜早破（t PROM）30例（t PROM组）,正常足月妊娠30例（正常组）。三组均于剖宫产术中胎盘娩出后于距离胎膜破口〉2 cm处取1 cm×1 cm的胎膜组织制作切片。采用HE染色方法检测绒毛膜羊膜炎（中性粒细胞中、重度浸润）发生情况;采用免疫组化SP法检测MIF、iNOS表达;分析三组及绒毛膜羊膜炎、非绒毛膜羊膜炎产妇MIF、iNOS表达情况。结果三组共检出绒毛膜羊膜炎27例;MIF和iNOS主要表达于羊膜上皮细胞的胞质。p PROM组和t PROM组MIF表达强度明显高于对照组（P〈0.01）;p PROM组iNOS表达强度明显高于对照组（P〈0.01）;绒毛膜羊膜炎产妇胎膜中MIF和iNOS表达强度明显高于非绒毛膜羊膜炎产妇（P〈0.05）。结论 PROM产妇胎膜组织中MIF、iNOS表达上调。MIF、iNOS高表达可促进胎膜早破及绒毛膜羊膜炎的发生。     

Layered hydrogels accelerate iPSC-derived neuronal maturation and reveal migration defects caused by MeCP2 dysfunction

Probing a wide range of cellular phenotypes in neurodevelopmental disorders using patient-derived neural progenitor cells (NPCs) can be facilitated by 3D assays, as 2D systems cannot entirely recapitulate the arrangement of cells in the brain. Here, we developed a previously unidentified 3D migration and differentiation assay in layered hydrogels to examine how these processes are affected in neurodevelopmental disorders, such as Rett syndrome. Our soft 3D system mimics the brain environment and accelerates maturation of neurons from human induced pluripotent stem cell (iPSC)-derived NPCs, yielding electrophysiologically active neurons within just 3 wk. Using this platform, we revealed a genotype-specific effect of methyl-CpG-binding protein-2 (MeCP2) dysfunction on iPSC-derived neuronal migration and maturation (reduced neurite outgrowth and fewer synapses) in 3D layered hydrogels. Thus, this 3D system expands the range of neural phenotypes that can be studied in vitro to include those influenced by physical and mechanical stimuli or requiring specific arrangements of multiple cell types.Neuronal migration and maturation is a key step in brain development. Defects in this process have been implicated in many disorders, including autism (1) and schizophrenia (2). Thoroughly understanding how neural progenitor cell (NPC) migration is affected in neurodevelopmental disorders requires a means of dissecting the process using cells with genetic alterations matching those in patients. Existing in vitro assays of migration generally involve measurement of cell movement across a scratch or gap or through a membrane toward a chemoattractant in 2D culture systems. Although widely used, such assays may not accurately reveal in vivo differences, as neuronal migration is tightly regulated by physical and chemical cues in the extracellular matrix (ECM) that NPCs encounter as they migrate.In vitro 3D culture systems offer a solution to these limitations (3–7). Compared with 2D culture, a 3D arrangement allows neuronal cells to interact with many more cells (4); this similarity to the in vivo setting has been shown to lengthen viability, enhance survival, and allow formation of longer neurites and more dense networks in primary neurons in uniform matrices or aggregate culture (8, 9). Indeed, 3D culture systems have been used to study nerve regeneration, neuronal and glial development (10–12), and amyloid-β and tau pathology (13). Thus, measuring neuronal migration through a soft 3D matrix would continue this trend toward using 3D systems to study neuronal development and pathology.We sought to develop a 3D assay to examine potential migration and neuronal maturation defects in Rett syndrome (RTT), a genetic neurodevelopmental disorder that affects 1 in 10,000 children in the United States and is caused by mutations in the X-linked methyl-CpG-binding protein-2 (MECP2) gene (14). Studies using induced pluripotent stem cells (iPSCs) from RTT patients in traditional 2D adherent culture have revealed reduced neurite outgrowth and synapse number, as well as altered calcium transients and spontaneous postsynaptic currents (1). However, 2D migration assays seemed unlikely to reveal inherent defects in this developmental process, which could be affected because MeCP2 regulates multiple developmental related genes (15). Migration of RTT iPSC-derived NPCs has not previously been studied.Using a previously unidentified 3D tissue culture system that allows creation of layered architectures, we studied differences in migration of MeCP2-mutant iPSC-derived versus control iPSC-derived NPCs. This approach revealed a defect in migration of MeCP2-mutant iPSC-derived NPCs induced by either astrocytes or neurons. Further, this 3D system accelerated maturation of neurons from human iPSC-derived NPCs, yielding electrophysiologically active neurons within just 3 wk. With mature neurons derived from RTT patients and controls, we further confirmed defective neurite outgrowth and synaptogenesis in MeCP2-mutant neurons. Thus, this 3D system enables study of morphological features accessible in 2D system as well as previously unexamined phenotypes.     

Blood Polymorphonuclear Leukocyte Activation in Atherosclerosis: Effects of Aspirin

The aim of the study was to demonstrate an activation of polymorpho-nuclear leukocytes (PMNs) in chronic progressive atherosclerosis (ATH). A group of patients with ATH, and a group of ATH patients under aspirin (ASA) therapy were compared with control persons without atherosclerotic alterations (healthy controls). Each group comprised 15 male age-matched subjects. The following inflammatory parameters related to PMN activities were measured: the polymorphonuclear leukocyte (PMN) blood count; blood PMN migration and reactive oxygen species release in vitro; the blood levels of PMN elastase, malondialdehyde, antibodies to oxidized LDL and soluble ICAM-1. In ATH patients, the PMN blood counts and the share of blood PMNs migrating upon platelet activating factor and leukotriene B4 stimulation were significnatly above the values of the healthy controls, while the other parameters were not significantly altered. ASA treatment attenuated the inflammatory response and reduced the differences between ATH and the healthy controls. It can be concluded that, in patients with chronic progressive atherosclerosis, PMNs are involved in the inflammatory process underlying the disease.     

巨噬细胞移动抑制因子在前列腺癌中的表达及其临床意义探讨

目的:探讨前列腺癌巨噬细胞移动抑制因子(MIF)的表达及其临床意义。方法:采用ELISA法检测36例前列腺癌、32例良性前列腺增生(BPH)及20例健康成年男性的血清MIF水平,并做统计学分析。结果:MIF在前列腺癌组血清中的水平明显高于BPH组和正常对照组,差异有统计学意义(P<0.05)。血清MIF水平与临床分期和Gleason评分明显相关(P>0.05)。结论:MIF在前列腺癌、侵袭过程中起着重要的作用,可能作为前列腺癌早期诊断和预测疾病进展的生物标记物之一。     

CXCR4/SDF-1α信号对乳腺癌细胞的体外增殖和迁移的促进作用

目的探讨CXCR4/SDF-1α信号在乳腺癌细胞体外增殖和迁移中的作用。方法免疫组织化学染色法检测CXCR4在乳腺癌组织中的表达;采用免疫荧光标记法和RT-PCR方法检测CXCR4在乳腺癌细胞株上的表达;MTT法研究SDF-1α细胞因子对乳腺癌细胞株体外增殖的影响及抗体12G5的阻断作用;体外微孔隔离室迁移技术研究CXCR4/SDF-1α信号对乳腺癌细胞株体外迁移能力的影响和抗体12G5的阻断作用。结果 CXCR4表达于乳腺癌组织,也在乳腺癌细胞株表达。SDF-1α细胞因子可有效促进乳腺癌细胞株MDA-M231的体外增殖和迁移,而阻断型CXCR4单抗12G5可有效抑制其增殖和迁移,并呈浓度依赖性。结论 CXCR4/SDF-1α信号参与了乳腺癌细胞株的体外生长和转移,与乳腺癌的侵袭和转移有关。