胚胎早期乳腺原基体外培养对乳腺干细胞相关标记表达稳定性的影响

乳腺干细胞具有产生各类乳腺细胞的能力,其研究对于乳腺发育的认知、乳腺癌的治疗以及乳腺生物反应器的开发都具有重要意义。此外,伴随胎儿发育,胚胎期乳腺干细胞所经历的增殖、迁移、侵袭等过程是成体乳腺干细胞没有的,而这些经历与乳腺癌的发生过程非常相似。因此,针对胚胎乳腺干细胞的研究在医学上具有更重要的意义。Sca-1、CD29、CD49f和CD24都是经常被用于乳腺干细胞分离纯化的细胞表面标记。该研究分析了不同的体外处理条件和时间对于上述标记在小鼠乳腺原基中表达稳定性的影响,旨在为胚胎早期乳腺干细胞的分离、纯化以及鉴定奠定分子基础,也为胚胎乳腺发育和乳腺癌发生的相关研究提供支持。     

识别宿主细胞黏附基质分子的细菌表面组分

许多细菌表面具有能识别并黏附于宿主细胞外基质(如纤连蛋白、胶原、纤维蛋白原等)的表面成分,统称为识别黏附基质分子的微生物表面组分。本文就其性质、结构,以及它们在细菌致病过程中的作用作一综述。     

突触细胞黏附分子在应激中作用的研究进展

突触细胞黏附分子是一类介导突触前、后膜结构和功能互作的膜表面糖蛋白,可以动态调节突触活动和可塑性,其表达与功能受到环境因素调控。突触细胞黏附分子亦是应激反应重要的效应分子之一,可介导应激对认知和情绪的不良影响。本文综述近年来突触细胞黏附分子在应激中作用的研究进展,旨在为应激相关障碍的分子机制研究和药物研发提供思路。     

肠道菌群与肠易激综合征分型及患者血清细胞间黏附分子 1表达的关系

目的分析肠道菌群与IBS分型及患者血清细胞间黏附分子 1(ICAM 1)表达的关系,为后续研究提供参考。方法选择2018年12月至2020年1月我院收治的152例符合标准的肠易激综合征(IBS)患者作为观察组,并依照罗马Ⅲ标准将观察组患者分为腹泻型组(62例)、便秘型组(56例)、混合型组(34例);选择同期我院健康体检者30例为对照组。检测受试者粪便中双歧杆菌、乳杆菌、肠杆菌和肠球菌数量,同时检测受试者血清ICAM 1水平。采用Pearson相关性分析患者肠道菌群与血清ICAM 1关系,并采用Logistics回归模型分析肠道菌群与IBS各亚型的关系。结果观察组患者血清ICAM 1、肠道双歧杆菌、乳杆菌水平显著低于对照组,肠道肠杆菌和肠球菌水平显著高于对照组,差异均有统计学意义(均P<0.05)。不同亚型IBS患者血清ICAM 1、肠道双歧杆菌、乳杆菌、肠杆菌和肠球菌水平存在明显差异,其中混合组患者血清ICAM 1、肠杆菌、肠球菌数量最高,双歧杆菌、乳杆菌数量最低;腹泻型组肠道肠杆菌、肠球菌数量最低,双歧杆菌、乳杆菌数量最高(均P<0.05)。肠道中双歧杆菌、乳杆菌、肠杆菌、肠球菌数量是IBS分型的独立影响因素(均P<0.05)。患者肠道双歧杆菌、乳杆菌数量与ICAM 1水平呈显著负相关,而肠道肠杆菌和肠球菌数量与ICAM 1水平呈显著正相关(均P<0.05)。结论肠道菌群可影响IBS患者分型,且肠道中部分菌群与ICAM 1水平存在显著相关关系。     

肺炎链球菌上调THP-1细胞分泌细胞间黏附分子1

目的 研究THP-1细胞受肺炎链球菌(SP)刺激后细胞间黏附分子1(ICAM-1)分泌水平的变化,并探讨不同来源SP对此影响的差异.方法 从温州医学院附属第二医院住院患者分离、搜集23株SP,和ATCC49619一起进行培养,并调浓度至1.0 McFarland.收集经传代、培养的THP-1细胞,调浓度至4.0×108/L,取1.0 mL加入24孔细胞培养板中,再向其中加入100 μL浓度为1.0 McFadand的SP,置37℃5％CO2环境分别孵育4和8h,吸出细胞悬液,离心取上清液.以ELISA法检测上清液中ICAM-1的浓度.以SPSS 17.0软件对检测结果进行统计分析.结果 SP刺激THP-1细胞4、8h后ICAM-1的浓度均高于相应的空白对照.血源性SP刺激THP-1细胞4、8h后ICAM-1的浓度与相应的痰源性SP间差异均无统计学意义.血源性/痰源性SP刺激THP-1细胞8h后ICAM-1的浓度均高于其刺激4h后的浓度.血源性/痰源性SP刺激THP-1细胞4h后ICAM-1的浓度均高于ATC C49619菌株刺激4h后相应的浓度,而在刺激8h后二者间差异无统计学意义.结论 SP可使THP-1细胞分泌ICAM-1增加,但其浓度变化与刺激的时间有关.血源性和痰源性SP刺激THP-1细胞分泌ICAM-1的变化比较差异无统计学意义,而二者和ATCC49619菌株间差异有统计学意义.     

小鼠FAAP蛋白对细胞黏附的影响

黏着斑相关蛋白(focal adhesion associated protein,FAAP)由小鼠D10Wsu52e基因编码,在进化上十分保守,但该蛋白质的生物学功能并不清楚．为此,首先通过细胞组分分离的方法研究了FAAP蛋白分布的细胞组分,结果表明FAAP主要存在于细胞质和细胞膜中．细胞蛋白表达量分析表明,细胞中FAAP与黏连蛋白受体(LR)表达量呈现正相关性．同时,细胞黏附实验表明,FAAP与LR对细胞黏附影响类似,也能够抑制细胞的黏附．这些实验结果为深入研究FAAP蛋白功能提供了依据．     

盘基网柄菌细胞黏附分子cadA/DdCAD-1在细胞分化及细胞命运决定中的作用

盘基网柄菌细胞黏附分子DdCAD-1是在细胞发育过程中最先表达的黏附分子,为了研究DdCAD-1在盘基网柄菌细胞发育中的作用,将cadA基因的突变株cadA-细胞用中性红染料染色,发育成的蛞蝓体显示cadA-细胞的前柄细胞/前孢子细胞的分化出现明显的障碍,外源表达的重组蛋白His6-DdCAD-1与cadA-细胞作用一段时间后,这种现象得到了改善。另外,cadA-细胞的孢子产率也有所降低,外源重组蛋白也可以拯救该表现型。表达DdCAD-1的细胞与cadA-细胞共同发育所形成的嵌合体显示表达DdCAD-1的细胞占据在拔顶期结构的顶端及尾部,而这些结构都在非孢子区,最终会死亡。提示DdCAD-1对于细胞分化及细胞命运决定有重要意义。     

黄芪对急性白血病患者血清黏附分子水平影响的临床研究

目的:探讨黄芪对急性白血病病人可溶性细胞间黏附分子-1和可溶性血管细胞黏附分子-1水平的影响。方法:64例初治急性白血病患者随机分为化疗组32例和化疗加黄芪组32例,采用酶联免疫吸附测定方法(EuSA法),对治疗前后的血清可溶性细胞间黏附分子-1和可溶性血管细胞黏附分子-1水平进行检测。结果:①与正常组比较,急性白血病病人治疗前后可溶性细胞间黏附分子-1和可溶性血管细胞黏附分子-1水平升高(P<0 05)。②治疗后,化疗加黄芪组与化疗组血清可溶性细胞间黏附分子-1和可溶性血管细胞黏附分子-1水平均下降(P<0 05),化疗加黄芪组下降尤为明显(P<0 05)。结论黄芪可通过降低白血病血清sICAM-1和sVCAM-1水平的而发挥抗肿瘤作用。     

上皮细胞黏附分子增强细胞间紧密连接促进乳腺癌细胞耐药

乳腺癌是致死率很高的恶性肿瘤,由ABCG2 (ATP-binding cassette G2)介导的多药耐药(multidrug resistance,MDR)是导致其化疗失败的重要原因,探讨ABCG2介导的耐药机制并探寻其关键分子是当前亟待解决的难题。上皮细胞黏附分子(epithelial cell adhesion molecule,EpCAM)参与多种肿瘤耐药,且与乳腺癌MDR密切相关,但它在ABCG2介导的乳腺癌耐药中的作用尚未阐明。本研究目的在于探究EpCAM对于ABCG2介导的乳腺癌细胞的多药耐药的调节作用及其机制。CCK8细胞毒性结果证实,相对于人乳腺癌药物敏感株MCF-7,耐药株MCF-7/MX对米托蒽醌(mitoxantrone,MX)的耐药性显著增强;Western 印迹结果显示,与MCF-7相比,MCF-7/MX细胞中ABCG2高表达,EpCAM表达上调。siRNA法敲低MCF-7/MX细胞中EpCAM可下调其ABCG2表达,并恢复对MX的敏感性。倒置显微镜观察细胞形态,发现敲低EpCAM可减少MCF-7/MX细胞间连接。免疫荧光双染法观察到EpCAM与密封蛋白1(claudin 1)在MCF-7/MX细胞共定位;进一步Western 印迹结果表明,敲低EpCAM减少MCF-7/MX细胞中密封蛋白1表达。综上所述,EpCAM可能通过与密封蛋白1相互作用,增强细胞间紧密连接,促进ABCG2介导的乳腺癌多药耐药。     

血管细胞黏附分子-1对卵巢癌细胞凋亡的影响及机制研究

目的:探讨过表达血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)对卵巢癌细胞凋亡的影响。方法:构建过表达VCAM-1的慢病毒载体GV358-VCAM1+,转染人类卵巢癌IGROV1细胞株,利用嘌呤霉素筛选稳定表达VCAM-1的IGROV1细胞,通过倒置荧光显微镜下观察绿色荧光,确定细胞转染效率,Western blot及RT-PCR法确定卵巢癌细胞VCAM-1蛋白和m RNA水平;采用流式细胞仪检测过表达VCAM-1的IGROV1的细胞凋亡变化,western blot法检测凋亡相关蛋白(Bcl-2、Bax、Casepase-3、Cleaved Casepase-3)以及STAT3、p-STAT3蛋白表达水平的变化。结果:成功构建的慢病毒载体GV358-VCAM1+在IGROV1细胞中的转染效率达到85%以上,转染细胞的VCAM-1蛋白及m RNA水平均呈稳定表达;VCAM-1过表达卵巢癌细胞的细胞凋亡显著高于空载体对照组(P=0.0149);Bax、Casepase-3、Cleaved Casepase-3表达水平均较对照组显著升高(P0.01),Bcl-2、p-STAT3表达水平明显低于对照组(P0.01),但STAT3表达水平无显著改变。结论:VCAM-1可能通过下调STAT3的磷酸化水平诱导卵巢癌细胞凋亡。     

Melatonin is Involved in Regulation of the Expression of Neural Cell Adhesion Molecules in the Rat Brain

Cell adhesion molecules play a diverse role in neural development, signal transduction, structural linkage to extracellular and intracellular proteins, synaptic stabilization, neurogenesis, and learning. Neural cell adhesion molecules (NCAM) are members of the immunoglobulin superfamily and are involved in synaptic rearrangements in the mature brain. There are three major NCAM isoforms: NCAM 180, NCAM 140, and NCAM 120. Several studies reported that NCAM play a central role in memory formation. We investigated the effects of melatonin on the expression of NCAM in the hippocampus, cortex, and cerebellum of rats. The levels of NCAM isoforms were determined by Western blotting. After administration of melatonin for 7 days, the expression of NCAM 180 increased both in the hippocampus and in the cortex, as compared with the control. In contrast, in rats exposed to constant illumination for 7 days (a procedure that inhibits endogenous production of melatonin), levels of NCAM 180 dropped in the hippocampus and became undetectable in the cortex and cerebellum. Levels of NCAM 140 in the hippocampus of light-exposed rats also decreased. There was no change in the expression of NCAM 120 in any brain region. This is the first report indicating that melatonin exerts a modulatory effect on the expression of NCAM in brain areas related to realization of cognitive functions. Melatonin may be involved in structural remodeling of synaptic connections during memory and learning processes.     

Heterogeneity of Soluble Neural Cell Adhesion Molecule

Soluble neural cell adhesion molecule (NCAM) from rat brain neuronal cell culture media consists predominantly of a polypeptide of Mr approximately 115,000. Minor amounts of a polypeptide of Mr approximately 180,000 and two inconsistently appearing components of Mr 160,000 and 145,000 are also observed. The Mr 115,000 component is derived from the neuronal membrane NCAM components NCAM-A of Mr 190,000, NCAM-B of Mr 140,000, or both. Thus, as a part of the catabolism of membrane NCAM-A plus -B, a minor fraction is posttranslationally cleaved and recovered in the media as discernible soluble NCAM polypeptides. The half-life of membrane NCAM-A plus -B is less than 24 h. Astrocyte culture media contains a predominant soluble NCAM component of Mr 120,000 derived from membrane-associated NCAM-C. A close comparison of deglycosylated soluble NCAM from astrocyte and neuronal cultures showed a small but consistent difference in Mr, a result suggesting that different NCAM polypeptides are released from the membrane of neurons and astrocytes. In contrast to the Mr 115,000-120,000 NCAM polypeptides, the Mr 180,000 polypeptide from neuronal culture media does not seem to be derived from membrane-attached NCAM and may therefore represent a secreted NCAM isoform.     

Developmental Expression of Neural Cell Adhesion Molecules of Oligodendrocytes In Vivo and in Culture

Abstract: Previously, we have shown that oligodendrocyte adhesion molecules are related to the 120,000–M_r neural cell adhesion molecule (NCAM-120). In this report, we present further evidence that the oligodendrocyte adhesion molecule is NCAM-120. Studies on the expression of NCAM-120 and other molecular forms of NCAM in vivo in rat brain, in vitro in primary mixed cultures, and in cultures enriched for oligodendrocytes are described. Western blot analysis of rat brain using anti-NCAM showed that NCAM-120 first appears at postnatal day 7 and increases in quantity thereafter, coincident with the development of oligodendrocytes in vivo and comparable to the expression of myelin basic protein. Purified oligodendrocytes from 4-week-old rat brains expressed only NCAM-120. Quantitation of various forms of NCAMs in rat brain showed marked age-related differences in the expression of three molecular forms of NCAM. Immunofluorescence analysis showed that oligodendrocytes, at all ages tested, expressed NCAM, but in older oligodendrocytes, the intensity of staining was less. Western blot analysis of oligodendrocyte-enriched cultures showed that from day 1 after isolation (12 days of age) through day 7 after isolation (18 days of age) only NCAM-120 is seen. A possible role for NCAM in myelination and remyelination is discussed.     

IFN—α对人脐带间充质干细胞粘附分子表达的影响

目的：本研究旨在观察不同浓度IFN-α对体外培养人脐带问充质千细胞表面粘附分子表达变化的影响。方法!采用组织块移行法培养人脐带间充质干细胞（Humanumbilicalcordmesenchymalstemcells,hucMSCs）,并进行干细胞表面抗原、成骨和成脂鉴定。向P3代hucMSCs加入不同浓度的IFN-α,24小时后收集细胞,应用流式细胞仪检测CD44、CD49d、CD54、CD58、CD62p、CD62L、CD102及CD106等八种粘附分子的表达情况。结果：①生理状态下,CD106、CD62P、CD62L和CD102阳性表达率极低（均〈1％）,CD54表达最高,为41．58％,经IFN-α干预后,CDl02、CDl06、CD62L、CD62p阳性表达率略有升高,但总体变化不明显（均〈5％）。②CD49d、CD54、CD58阳性表达率与IFN-α呈浓度依赖性,最高达（66．36＋2．48）％、（76．26＋1．85）％、（47．78＋O．44）％;CD44在浓度为3×103U／ml时阳性表达率最高,为（49．81±3．25）％,且干预组与对照组、各组间比较差异有显著性意义（P〈0．05）。结论：炎症因子IFN-α可显著提高hucMSCs表面CD54、CD58、CD44、CD49d的阳性表达率,但对CD102、CD106、CD62P和CD62L作用不明显。     

Characterisation of Antibodies Specific for Chick Brain Neural Cell Adhesion Molecules Which Cause Amnesia for a Passive Avoidance Task

Abstract: Antisera were prepared against six postsynaptic density glycoprotein fractions (150–180, 62–80, 50, 41, 33, and 28 kDa) that show enhanced fucosylation during memory formation after training day-old chicks in a one-trial passive avoidance task. Each antiserum was tested for its possible effect on memory retention. Bilateral intracranial injections of two of the antisera, R-1 and R-6, or their IgGs (IgG-1 and IgG-6), resulted in amnesia for the passive avoidance task when chicks were tested 24 h later. IgG-1 and IgG-6 antibodies were amnestic only when injected 5.5 h after training, and had no effect when injections were made 30 min before training, thus resembling an effect previously observed with polyclonal or monoclonal anti-N-CAM antibodies. IgG-1 and IgG-6 antibodies were found to be specific for protein epitopes of glycoproteins that contain a high amount of N-linked mannose and fucose, and a very low amount of polysialic acid and O-linked galactose. Absorption of IgG-6 antibodies with neural cell adhesion molecule (N-CAM) isolated from synaptic plasma membranes derived from day-old chick brain resulted in loss of amnestic effect. As we have previously shown that long-term memory for the passive avoidance task requires two waves of glycoprotein synthesis, the first occurring immediately after training and the second 5–8 h later, the present results suggest strongly that isoforms of N-CAM molecules with a low level of sialic acid are involved specifically in the establishment of an enduring memory for the experience of the passive avoidance task in chicks, possibly by stabilising changes in synaptic connectivity that encode the memory.     

Glucocorticoids and Nerve Growth Factor Differentially Modulate Cell Adhesion Molecule L1 Expression in PC12 Cells

Abstract: The differential expression of the cell adhesion molecule L1 by chromaffin cells has recently been suggested to be responsible for the segregation of chromaffin cells into homotypic catecholaminergic groups in the adrenal gland. The present study was undertaken to test the hypothesis that glucocorticoids, which increase in the adrenal gland during development, could be responsible for the repression of L1 in adrenergic chromaffin cells. PC12 cells were used as the experimental model, and relative L1 protein and mRNA levels were examined after treating the cells with glucocorticoids or NGF. Analysis of western blots indicated that glucocorticoids decreased the L1 protein levels by one-half, whereas NGF increased L1 protein levels ∼2.3-fold. In addition, the glucocorticoids inhibited both the NGF induction of the neurite outgrowth and the increase in L1 expression. Analysis of the mRNA levels by PCR and northern blots indicated that glucocorticoids reduced the L1 mRNA, whereas NGF increased the level of L1 mRNA. Maximal inhibition of L1 expression was observed at concentrations of 10⁻⁷ M dexamethasone, and the decrease occurred during the second day of treatment. The effects of dibutyryl cyclic AMP and phorbol ester on the glucocorticoid and NGF regulation of L1 protein were also examined. This is the first report indicating that L1 expression can be down-regulated by glucocorticoids. The results support the hypothesis that during development the repression of L1 in adrenergic chromaffin cells may be, in part, linked to the increase in glucocorticoid levels in the adrenal gland.     

Transplantation of a Mammary Stromal Cell Line into a Mammary Fat Pad: Development of the Site-Specific in Vivo Analysis System for Mammary Stromal Cells

The interaction between mammary epithelial and stromal tissue is considered to be important in breast tissue development. In this study, we developed a transplantation procedure for the mammary stromal fibroblastic cell line (MSF) to examine its life in vivo. First we established MSF cells which stably expressed lacZ (lacZ/MSF) and had characteristics of mammary stromal cells. The lacZ/MSF cells were then transplanted into a cleared mammary fat pad of syngenic mice with and without mammary primary epithelial organoids. Whole mount X-gal and carmine staining of the transplants revealed that a number of undifferentiated lacZ/MSF cells survived around the mammary epithelial tissue when transplanted with organoids. These results indicate that transplantation of MSF cells into mammary fat pad was accomplished by co-transplantation with primary mammary organoids. Finally, we discuss the application of transplantation procedure for in vivo studies of the mammary stromal tissue development and stromal-epithelial interactions.     

Expression of Cell Adhesion Molecules and Cytokines in Murine Antigen-Induced Arthritis

Adhesion molecules and cytokines are important in chronic inflammatory conditions such as rheumatoid arthritis (RA) by virtue of their role in cell activation and emigration. Using immunohistochemical techniques we studied the expression of adhesion molecules and cytokines in cryopreserved sections of murine knee joint in the course of antigen-induced arthritis, an animal model of human RA. Various adhesion molecules and cytokines are expressed in the arthritic joint tissue. LFA-I, Mac-1, CD44, ICAM-I and P-selectin were strongly expressed in the acute phase and to a lesser degree in the chronic phase of arthritis. VLA-4 and VCAM-I appeared to be moderately expressed on day 1, L-selectin between days 1 and 3. LFA-I, Mac-I, CD44, a4-integrin, ICAM-I and the selectins were found expressed on cells of the synovial infiltrate, LFA-1, Mac-1 and ICAM-I on the synovial lining layer, and VCAM-I and P-selectin on endothelial cells. Expression of E-selectin could be demonstrated throughout the experiment at a low level in cells of the acute cell infiltrate. Cytokines, especially IL-2, IL-4, IL-6, TNF, and IFN-7, were heavily expressed during the acute phase of arthritis in cellular infiltrate. Taken together these data demonstrate that cytokines and their activation of adhesion molecules contribute to cell infiltration and activation during the initial phase of arthritis and to the induction and progression of tissue destruction in arthritic joints. These molecules might be potential targets for novel therapeutic strategies in inflammatory and arthritic disorders.     

Equilibrium Binding Analysis of Neural Cell Adhesion Molecule Binding to Heparin

The kinetics of neural cell adhesion molecule (NCAM) binding to heparin were studied in a heparin-Sepharose-based solid-phase binding assay. The observed binding is time dependent and saturable. A binding constant of 5.2 +/- 1.4 X 10(-8) M is observed for binding of newborn rat NCAM to heparin. This is approximately 25 times lower than the binding constant determined for newborn rat NCAM homophilic binding. Both Scatchard and Hill plot analyses suggest the presence of only one binding site. Fab' fragments of antibodies to rat NCAM significantly inhibit binding, a result indicating that a specific site on NCAM is involved in binding to heparin. The binding is inhibited by heparin (IC50, approximately 5 micrograms/ml), whereas chondroitin sulfate is a less potent inhibitor (IC50, approximately 15 micrograms/ml).