大肠癌Lovo细胞E-Cadherin,N-Cadherin的表达及化学治疗药物对其表达的影响

目的：研究大肠癌Lovo细胞上皮钙粘附素(E-Cadherin)，神经钙粘附素(N-Cadherin)的表达及常用化学治疗药物对其表达的影响。方法：应用逆转录聚合酶链反应(RT-PCR)法检测大肠癌Lovo细胞E-Cad-herin，N-Cadherin的表达，并通过几种不同的化疗药物(顺铂、吡柔比星、丝裂霉素、5-FU)不同浓度和时间作用后，对E-Cadherin，N-Cadherin表达的影响。结果：不同化疗药物对大肠癌Lovo细胞N=Cadherin表达无影响，高浓度顺铂和高浓度吡柔比星二组出现-Cadherin表达，其它各组未见表达。结论：大肠癌常用的化疗药物无论低浓度持续用药，还是高浓度短时间用药，对大肠癌Lovo细胞N-Cadherin的表达无抑制作用，说明上述药物不能通过抑制N—Cadherin的表达，而起到抗癌作用。高浓度顺铂和吡柔比星二组显示E-Cadherin的表达，表现出抑癌的作用，从而提示在大肠癌化疗时，顺铂和吡柔比星更适合于高浓度短时间应用。     

乳腺癌淋巴道转移潜能与癌细胞中钙粘素及金属蛋白酶的关系

背景及目的:目前尚缺少有效评价乳腺癌淋巴道转移潜能的理想指标。本研究旨在通过检测上皮性钙粘素(E-cadherin,E-Cad)、神经性钙粘素(N-cadherin,N-Cad)和基质金属蛋白酶-9(matrixmetalloproteinase-9,MMP-9)基因产物在乳腺癌组织中表达的情况来探讨它们与乳腺癌浸润和转移的关系。方法:采用免疫组织化学SP方法检测E-Cad、N-Cad和MMP-9在72例乳腺浸润癌(其中淋巴结转移39例,无淋巴结转移33例)中的表达,并用多因素Cox比例风险模型分析患者的预后。结果:E-Cad表达在淋巴结转移组和无淋巴结转移组乳腺癌肿瘤细胞中的平均秩次分别为29.19,45.14,两组差异显著(P<0.001),E-Cad表达与乳腺癌转移呈负相关;N-Cad和MMP-9在淋巴结转移组乳腺癌细胞中的平均秩次分别为40.04和42.97;在无淋巴结转移组乳腺癌肿瘤细胞中的平均秩次分别为32.32和28.85。二者在淋巴结转移组与无淋巴结转移组的表达均具有显著性差异(P<0.05),与乳腺癌淋巴结转移呈正相关。E-Cad高表达者生存时间长。结论:乳腺癌的淋巴道转移与E-Cad、N-Cad和MMP-9的表达具有显著相关性,检测这几种蛋白表达将有助于判断乳腺癌的转移潜能及预后。     

Effect of artesunate on human endometrial carcinoma HEC-1B cells

Objective： To observe the effect of the artesunate （ART） on cellular proliferation in vitro, to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the experimental and theoretical foundations for the clinical applications of ART. Methods： The cell proliferation was observed by microscope; MTT was used to examine the effects of ART on proliferation of HEC-1B cells, and flow cytometric analysis was used to detect cell cycle and apoptosis. The human endometrial carcinoma HEC-1B cells were conventionally cultured; ART was administered with a concentration of 40 μg/ml before the total RNA were extracted, mRNA expression of Survivin, Caspase-3, N-Cadherin, E-Cadherin, Fibronectinl and Cox-2 were detected using RT-PCR. Results： ART reduced proliferation in human endometrial carcinoma cell line HEC-1B in a dose- and time-dependent effect. The cells of G0/G1 stage were significantly increased （P〈0.05）, but the cells of G2/M stages were significantly decreased （P〈0.05）, so it has shown that the cell cycle was probably blocked in G0/G1 stage. After intervention with ART at 20 and 80 μg/ml for 48 h, cellular apoptosis rate respectively was （36.42±0.77）% and （11.77±0.58）%, and the difference was statistically significant compared with the control （[6.64±0.191%, P〈0.01）. The expression of Cox-2 mRNA in the ART group was lower than those of control group, yet the expression of Caspase-3 and E-Cadherin mRNA in the ART group was higher than those of control group. Conclusion： ART can inhibit HEC-1B cell growth and proliferation in a dose- and time-dependent manner. Furthermore, ART can induce apoptosis in a dose-dependent manner. ART is able to downregulate Cox-2 mRNA expression and to upregulate E-Cadherin and Caspase-3 mRNA expression. So we can conclude that ART could induce the endometrial carcinoma HEC-1B cell apoptosis and inhibit tumor cell proliferation.     

上皮-间质转化相关标志物对鼻咽癌远期疗效预测的意义

目的探讨上皮-间质转化(epithelial-mesenchymal transition,EMT)相关标志物神经钙粘连素(N-Cadherin)、上皮钙粘连素(E-Cadherin)、波形蛋白(Vimentin)和Twist在鼻咽癌肿瘤组织中的表达,及其对鼻咽癌复发、转移和预后的意义。方法采用免疫组织化学方法检测77例鼻咽癌组织标本中的E-Cadherin、N-Cadherin、Vimentin和Twist蛋白表达。通过计算机辅助图像分析系统定量分析各EMT相关标志物的表达情况。采用单因素和多因素分析方法评估上述标志物对鼻咽癌复发、转移的预测价值。结果单因素分析显示,E-Cadherin、N-Cadherin、Vimentin和Twist蛋白表达与鼻咽癌复发、转移相关(P<0.05)。多因素分析显示,E-Cadherin和Vimentin是影响鼻咽癌复发、转移的独立风险因素。采用Kaplan-Meier法进行生存分析并作生存曲线。Log-rank检验显示,肿瘤组织内E-Cadherin、N-Cadherin、Twist低表达与高表达患者间生存率均有显著差异(P<0.05),而Vimentin高表达与低表达患者间生存率差异不显著(P=0.055)。结论 E-Cadherin、N-Cadherin、Vimentin和Twist蛋白与鼻咽癌转移、复发密切相关。EMT相关标志物对鼻咽癌侵袭、转移和预后有一定的预测价值。     

Cardiac cell-cell junctions in health and disease: Electrical versus mechanical coupling

Intercalated discs are the membrane sites where individual cardiomyocytes are connected to each other. Adherens-, desmosomal-, and gap junctions are situated in the intercalated disc and ensure mechanical coupling between cells and enable propagation of electrical impulses throughout the heart. A number of cardiac disorders, for example arrhythmogenic right ventricular dysplasia/cardiomyopathy, have been described in which an impaired mechanical coupling leads to electrical dysfunction, with occurrence of fatal arrhythmias.In this article the interaction between electrical and mechanical coupling is explored by reviewing studies performed in patients, animals, and in vitro. In these studies the effect of changes in protein composition of a mechanical junction on the electrical junction, and vice versa were investigated. It is shown that impaired electrical coupling does not change mechanical coupling. However, impaired mechanical coupling largely affects electrical coupling.     

鸡胚发育过程N-Cadherin和Neurofilament在视顶盖中的表达模式研究

目的:阐明鸡胚发育过程中神经钙黏蛋白(N-Cadherin)和神经丝蛋白(Neurofilament,NF)在视顶盖中的表达模式.方法:从鸡胚发育第6天(E6)开始到孵化的E20,分别取材、固定、包埋和冰冻切片,采用荧光免疫组织化学法检测N-Cadherin和NF蛋白在视顶盖各层的表达.结果:在鸡胚发育过程中,从E6开始,N-Cadherin和NF便呈强阳性表达,并且表现出明显各层分布的差异,E10-E14时,在NF强表达的区域与N-Cadherin重叠,E14之后NF的表达减弱,N-Cadherin无明显变化.结论:N-Cadherin和NF在鸡胚发育过程视顶盖的表达呈明显的层的分布,二者的表达既存在差异又有重叠.     

Transition of asthmatic bronchial fibroblasts to myofibroblasts is inhibited by cell-cell contacts

The role of airway wall remodelling in bronchial asthma is well established. Myofibroblasts, the cells displaying features intermediate between fibroblasts and smooth muscle cells, are involved in this process but the mechanism of myofibroblasts activation in the onset of the disease remains obscure. Myofibroblasts can differentiate from various cell types, including resident fibroblasts, and the fibroblasts to myofibroblasts transition (FMT) can be reproduced in?vitro. We aimed to investigate the process of FMT in human bronchial fibroblasts (HBF) derived from non-asthmatic (n?=?7) and asthmatic (n?=?7) subjects. We also tested whether cell-cell contacts affect FMT by using N-cadherin blocking antibody. HBF plated in low or high cell density were treated with TGF-β(1) up to one week to induce FMT. The percentage of myofibroblsts was counted and expression of α-smooth muscle actin was evaluated by cytoimmunofluorescence, flow cytometry and immunobloting. We demonstrated that the intensity of FMT induced by TGF-β(1)in?vitro was strongly enhanced in asthmatic as compared to non-asthmatic HBF populations. This process was facilitated by low cell plating density in both groups of cultures. Furthermore, we proved that neither HBF-conditioned medium nor growth arrest in G(0)/G(1) phase of cell cycle could stop the TGF-β(1)-induced FMT in asthmatic cell populations. However, even in sparse asthmatic HBF, the blocking of N-cadherin resulted in the inhibition of FMT. Our findings show for the first time that the initial absence or an induced loss of cell-cell adhesions in asthmatic HBF populations is important for the completion of FMT.     

小鼠胚胎发育过程中N-Cadherin异常表达对大脑皮层神经元迁移的影响

目的:探究N-Cadhetin异常表达对小鼠大脑皮层发育过程中神经元迁移的影响.方法:通过小鼠活体子宫电转基因技术和RNA干扰技术相结合,在小鼠胚胎大脑皮层发育过程中实现对N-Cadherin的超表达和抑制表达,荧光免疫组化检测超表达结果,用GFP示踪N-Cadherin超表达和抑制表达后神经元在大脑皮层发育过程中的迁移特征.结果:与对照组相比,N-Cadherin超表达后有更多的GFP阳性神经元迁移到皮层的Ⅱ-Ⅳ层;N-Cadherin被抑制后大量GFP阳性神经元停滞在SVZ区.结论:N-Cadherin在小鼠大脑皮层发育过程中对神经元的迁移具有影响作用.     

戊四氮癫痫幼鼠海马内NF-κB与N-Cadherin定位与苔藓纤维发芽现象

目的:探讨单次癫痫发作后脑内NF-κB和N-Cadherin表达与海马结构中苔藓纤维发芽现象之间的关系与作用。方法:利用腹腔注射戊四氮(PTZ)制作发育期SD大鼠(14 d、28 d)单次惊厥发作模型,各日龄组随机分为生理盐水(NS)组、PTZ组、吡咯二硫氨基甲酸酯(PDTC)组和PDTC+PTZ组,采用免疫组化法检测NF-κB和N-Cadherin的表达,利用Timm染色法观察海马苔藓纤维发芽现象。结果:NS组在海马CA3区可见极少Timm染色颗粒;致惊后1周偶见Timm染色颗粒、3周可见Timm染色颗粒沿海马CA3区呈条带样分布(P<0.01);PDTC预处理组Timm染色颗粒较PTZ致惊组明显减少(P<0.05)。NS组大鼠海马CA1、CA3区无NF-κB p65核转位细胞,致惊后24 h各日龄组幼鼠海马CA1、CA3区NF-κB p65核转位细胞较NS组明显增加(P<0.01);PDTC预处理后NF-κB p65的核转位细胞较致惊组明显减少(P<0.05)。NS组海马CA1、CA3区可见少量N-Cadherin阳性细胞;致惊组海马CA3和齿状回门区的N-Cadherin的阳性细胞与NS组相比明显增多(P<0.01),PDTC预处理后相同区域内N-Cadherin阳性细胞较PTZ致惊组明显减少(P<0.05)。NF-κB p65、N-Cadherin及Timm染色颗粒的表达结果与惊厥鼠的日龄并无关联性。结论:幼鼠单次惊厥发作可以引起海马不同程度的苔藓纤维发芽,而NF-κB p65、N-Cadherin的分布位置与变化时相与苔藓纤维发芽相吻合,表明NF-κB p65、N-Cadherin可能参与或伴随着苔藓纤维的发芽。