Assay of cell attachment and spreading factors

Summary Methods are presented for the quantitative assay of proteins influencing cell attachment and spreading. These include assays in which cell-substratum interactions are quantitated directly and a serum-free assay in which cell growth is dependent on attachment under substratum-limited conditions. Procedures are also presented for the identification of active sites of substratum molecules through the use of antibodies and synthetic peptides that are inhibitory for cell attachment.     

An ultrastructural and immunohistochemical study of microcystic meningioma with emphasis on matrix proteins and connexin 26 type gap junctions

Although the histopathological subtypes of meningioma do not themselves appear to have prognostic significance, they are collectively important for defining the overall histopathological entity of microcystic meningioma (MCM) and allowing a distinction from other intracranial tumors, such as capillary hemangioblastoma, glioma, and metastatic renal cell carcinoma showing similar histology. Four cases of MCM were analyzed by conventional histology, immunohistochemistry, and electron microscopy. The present series of MCM was characterized by spindle- or cobweb-shaped tumor cells, characteristically associated small blood vessels, and a peculiar microcystic pattern. Among the microcystic meningeal tumor tissue, small areas of conventional subtypes were identified. Immunohistochemically, tumor cells showed the mesenchymal features of vimentin positivity and a rich distribution of matrix proteins around tumor cells. They lacked epithelial marker positivity but were faintly EMA positive. Ultrastructurally, primitive cellular junctions, desmosomes, and gap junctions were frequently seen between tumor cells. The gap junctions correlated with connexin 26 immunoreactivity. Although lacking an obvious epithelial nature, these features could be interpreted as showing an abortive differentiation mimicking meningothelial (arachnoidal) cells, which, physiologically, regulate cerebrospinal fluid between blood vessels and brain parenchyma.     

The cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis

Ｌａｍｉｎｉｎ(ＬＮ)ｉｓａｍａｊｏｒｎｏｎｃｏｌｌａｇｅｎｏｕｓｇｌｙｃｏｐｒｏｔｅｉｎｉｎｂａｓｅｍｅｎｔｍｅｍｂｒａｎｅｓＩｔｈａｓｂｅｅｎｒｅｐｏｒｔｅｄｔｈａｔＬＮｃｏｕｌｄａｆｆｅｃｔａｎｕｍｂｅｒｏｆｂｉｏｌｏｇｉｃｂｅｈａｖｉｏｒｓｏｆｃａｒｃｉｎｏｍａｃｅｌｌｓｍｅｄｉａｔｅｄｂｙｔｈｅＬＮｒｅｃｅｐｔｏｒｓ1　ＩｔｉｓｋｎｏｗｎｔｈａｔｔｈｅＮｌｉｎｋｅｄｃｏｍｐｌｅｘｔｙｐｅｃａｒｂｏｈｙｄｒａｔｅｃｈａｉｎｓｏｎＬＮａｒｅｉｎｖｏｌｖｅｄｉｎｔｈｅｒｅｃｏｇｎｉｔｉｏｎａｎ…     

鼻咽癌中FN、LN和Col.IV的免疫组化研究

本文采有免疫组化方法用纤维连接蛋白（fibronectin,FN),层粘连蛋白（laminin,LN),和Ⅳ型胶原（collagenIV,Col,IV)抗体对54例鼻咽癌及1例鼻咽鳞状上皮瘤的基底膜进行了研究。结果显示鼻咽鳞状上皮瘤中的基底膜呈连续线状,而鼻咽癌的癌巢周围基底膜通常是不完整的,呈断续甚至消失的改变。高分化鳞癌显示连续线状基膜,偶有中断。低分化腺癌则显示断续不完整的基膜,大圆型细胞癌,低分化鳞癌部分显示连续线状基膜,偶有中断,未分化癌的基膜显示断线和消失的改变,癌瘤中基底膜的分布情况与癌实质类型,间质结构以及间质中淋巴细胞,浆细胞数量,嗜银纤维分布类型有关。     

血清Ⅲ、Ⅳ型胶原及板层素对肝纤维化的诊断价值

目的：探索诊断早期肝纤维化的敏感指标。方法：用放射免疫法检测２３６例各型肝病患者血清Ⅲ型胶原（ＴｐｙｅⅢｃｏｌａｇｅｎ,ＰＣⅢ）、Ⅳ型胶原（ＴｐｙｅⅣｃｏｌａｇｅｎ,Ⅳ－Ｃ）和板层素（Ｌａｍｉｎｉｎ,ＬＮ）水平。结果：各类肝病患者血清ＰＣⅢ、Ⅳ－Ｃ和ＬＮ水平随病情进展而逐渐升高。以ＰＣⅢ≥１３５μｇ／Ｌ、Ⅳ－Ｃ≥１３０μｇ／Ｌ为诊断肝纤维化的临界点,其敏感度、特异性、准确率分别达８０％以上。结论：表明ＰＣⅢ、Ⅳ－Ｃ及ＬＮ可作为肝纤维化诊断较为敏感的指标,其中Ⅳ－Ｃ对早期肝纤维化的诊断优于ＰＣⅢ及ＬＮ,对肝癌的诊断有一定意义     

Ⅳ型胶原,纤维连接蛋白和层粘连蛋白在舌癌中的表达和意义

探讨Ⅳ型胶原,纤维连接蛋白（ＦＮ）,层粘蛋白（ＬＮ）与舌鳞形细胞癌（ＴＳＣＣ）的关系,应用免疫组化ＳＰ法检测ＴＳＣＣ３０例中Ｃｏｌ Ⅳ,ＦＮ和ＬＮ的表达情况。结果：ＣｏｌⅣ、ＦＮ和ＬＮ表达一致,癌旁原位癌基底膜呈线状,偶见局部染色模糊或中断,浸润癌癌巢周围表达阳性２１例,阳性９例,阳性率在分化好,无颈淋巴结转移组明显高于分化差,颈淋巴结转移组;癌巢呈团块型者,癌团呈较完整线状表达,染色强度深浓,细     

层黏连蛋白及其受体在胃癌中的表达及其与侵袭和转移的关系

目的 :探讨层黏连蛋白 (LN)和LN受体 (LN R)的表达与胃癌侵袭和转移的关系。方法 :应用免疫组织化学S P法检测 68例胃癌组织中LN和LN R的表达。结果 :LN在有淋巴结转移组中阳性表达率明显低于无转移组 ,P <0 0 1;LN在其他各组中的表达差异无统计学意义 ,P >0 0 5 ;LN R的阳性表达率有随肿瘤分化水平的降低、浸润深度的增加而增高 ,P <0 0 1;淋巴结转移组中的LN R表达明显高于无淋巴结转移组 ,P <0 0 0 1。结论 :LN的低表达与肿瘤的转移有关 ,LN R的高表达与肿瘤分化、侵袭和转移的关系十分密切。LN和LN R可作为胃癌进展的重要生物学指标之一。     

层黏连蛋白受体反义核酸对卵巢癌细胞侵袭能力的影响

目的:研究67 kD层黏连蛋白受体(67kDLN-R)反义寡核苷酸(antisense oligonucleotide,ASODN)对卵巢癌细胞HRA体外侵袭转移能力的影响.方法:采用流式细胞仪、RT-PCR及Transwell小室的方法检测反义核酸对67 kD LN-R基因和蛋白表达影响,及转染前后细胞体外侵袭转移能力变化.结果:67 kD LN-R ASODN可从蛋白质和mRNA水平下调67kDLN-R的表达,且下调作用呈剂量依赖性.与正义组及对照组相比差异有显著性(P＜0.05).体外侵袭实验证实转染后HRA穿透人工基底膜的能力显著降低并呈现剂量依赖性.结论:67kDLN-R ASODN可降卵巢癌细胞体外侵袭转移能力,有望为卵巢癌治疗提供新方向.     

层粘连蛋白受体基因表达谱分析

目的 探讨层粘连蛋白受体（67LR1）基因相关的一组基因表达变化，为反式二羟环氧苯并芘致癌机制的研究提供依据。方法 构建67LR1转基因表达载体，转染人支气管上皮细胞（16HBE），获得转基因67LR1的瞬时表达的细胞株。用半定量RT-PCR方法分析转基因表达产物，应用含8064个基因点的微芯公司基因表达谱芯片CSC-GE-80与双荧光标记样品与芯片杂交和扫描、图像数据分析等一系列处理进行表达谱分析。结果 成功构建出瞬时表达67LR1的细胞株。基因表达谱分析，2组相比发生了显著性表达变化的基因有295个，其中表现为上调的基因有145个，下调表达的基因有150个。在发生了显著性表达改变的基因中，比较有明显功能分类特征的包括与信号转导相关的基因，与肿瘤相关的基因，与免疫反应相关的基因以及与蛋白质合成系统相关的基因等。结论 67LR1基因功能涉及到信号转导、肿瘤相关基因等方面，相关基因的表达变化具有复杂性。