细胞角蛋白的生物学功能

细胞角蛋白是细胞骨架蛋白的一部分，特定的表达于不同的上皮细胞及组织中，对细胞角蛋白表达的测定被广泛应用于检测肿瘤的来源。在此对细胞角蛋白其他重要的生物学功能做一综述，以期对临床工作提供理论指导依据。     

64例原发性肝癌角蛋白抗体标记观察

细胞角蛋白是上皮型细胞中丝蛋白的一种,它存在于绝大多数上皮细胞及其相应的肿瘤细胞中,因此用角蛋白制备的抗体可用于低分化恶性上皮性肿瘤的鉴别诊断.由于角蛋白种类多,不同种上皮所含     

乳腺增生性病变与乳腺癌的免疫组化研究

本文２４例乳腺良性增生性病变与１１例乳腺导管癌标本，采用细胞角蛋白，肌动蛋白和Ｓ－１００蛋白免疫组化染色。结果良性上皮细胞或癌细胞，细胞角蛋白均阳性，而肌上皮细胞阴性。肌动蛋白染色上皮和癌细胞均阴性，良性病变肌上皮细胞阳性。Ｓ－１００蛋白染色结果与肌动蛋白大致相仿，但反应较弱，结果提示鉴别乳腺良恶性病变以采用肌动蛋白为好。     

单(多)克隆抗体在肿瘤鉴别诊断中的应用(97例疑难病例分析)

所用一抗有角蛋白(Keratin),上皮细胞膜抗原(EMA)、细胞角蛋白(CK)、波状纤维(Viemutiu)、结蛋白(Desmin),α一抗胰蛋白酶(ACT)溶菌酶(Lysozyse)、白细胞共同抗原(LCA)、S-100蛋白、神经元特异性烯醇化酶(NSE)、嗜铬粒(Chromogla-     

细胞角蛋白7的表达在肿瘤及炎症中的意义

细胞角蛋白(Krt)作为一种中间丝,存在于所有的上皮细胞及部分非上皮细胞.主要作用是维持上皮细胞的机械稳定性和完整性.近年来细胞角蛋白的研究集中在肿瘤的诊断方面,即利用细胞角蛋白在上皮细胞表达的特异性,通过免疫组化技术,运用单克隆抗体进行比对分析,来确定肿瘤的分类、分型或者来源.其中Krt7是一个代表.探讨Krt7的作用,在上皮肿瘤中的分布,以及与细胞角蛋白20(Krt20)在上皮肿瘤中的联合表达情况,对肿瘤的诊断、转移性与原发肿瘤的鉴别等有重要意义.除此之外,Krt7还是一个炎性指标,发挥信号转导中的作用,参与炎症的发生发展,其表达也有利于对某些疾病早期及预后进行监测.     

结膜上皮细胞在角膜基质中转化为角膜上皮样细胞的研究

<正>结膜和角膜上皮细胞来源于不同的细胞系,正常情况下,分化成熟的结膜和角膜上皮细胞表达不同的角蛋白,即角膜上皮细胞表达CK3/CK12蛋白,而结膜上皮细胞表达CK4/CK13蛋白。而且,Ck12和Ck13分别代表终末分化的角膜和结膜上皮细胞,角膜缘干细胞或早期发育角膜细胞不表达CK4和CK13蛋白[1]。眼表上皮细胞分化除由其干细胞系所决定外,是否会受到其下基质环境的影响,即成体干细胞是     

鼻息肉上皮细胞角蛋白的表达及意义

对３８例鼻息肉石蜡及冰冻切片进行免疫组化染色，观察其上皮细胞角蛋白的表达，结果显示，细胞角蛋白可不同程度表达鼻息肉多种上皮细胞，提示鼻息肉粘膜上皮细胞反复变性，坏死，以及反复再生，分化的动态变化。     

借助自组装多肽水凝胶构建高活性复层结膜上皮细胞片

目的:评价RAD16-I自组装多肽水凝胶构建复层结膜上皮细胞片的效果,并探讨其作用机制.方法:酶消化法提取兔结膜上皮细胞(rCECs),倒置显微镜观察其形态,阿尔辛蓝-过碘酸-希夫染色检测杯状细胞,免疫荧光检测特异性蛋白细胞角蛋白4(CK4)和黏蛋白5AC(MUC5AC)的表达;磷脂酶脱细胞法制得脱细胞的细胞外基质(d...     

成人食管上皮细胞的体外培养及生物学特性

目的 培养成人正常食管上皮细胞,建立能够体外长期培养的食管上皮细胞系,为上皮细胞的体外研究提供实验材料.方法 取食管癌患者正常食管上皮,用0.25%Dispase酶和0.25%胰蛋白酶/0.02?TA消化获取成人食管上皮细胞,使用无血清角化细胞培养液培养,通过细胞形态学观察和角蛋白、上皮膜抗原(EMA)免疫组织化学染色鉴定细胞.结果 原代培养8d后,细胞汇合成片呈铺路石样生长,细胞角蛋白、上皮膜抗原表达阳性,可连续传代.结论 为体外分离培养成人正常食管上皮细胞建立了方便可行的方法.     

细胞角蛋白 19 通过调控 MAPK 和 PTEN 通路促进甲状腺癌细胞 的增殖、 侵袭及转移

目的 探究细胞角蛋白 19 对甲状腺癌细胞生物学过程的影响及相关机制。 方法 将人甲状腺癌 细胞 (human thyroid cancer cells, B-CPAP) 作为研究对象, 使用不同浓度的角蛋白 19 处理细胞, 通过 MTT 实验、 细胞划痕实验和肿瘤细胞侵袭实验探究细胞角蛋白 19 对 B-CPAP 增殖、 侵袭及转移的影响。 最后通 过研究相关蛋白分析细胞角蛋白 19 对 B-CPAP 的作用机制。 结果 MTT 实验发现, 随着角蛋白 19 浓度升 高, 其对 B-CPAP 的增殖率也明显提高。 Transwell 实验发现, 与空白对照组比较, 角蛋白 19 处理后通过小 室的细胞数明显增加。 细胞划痕实验发现, 角蛋白 19 能够促进 B-CPAP 的迁移能力, 并且 B-CPAP 的迁移 能力与角蛋白 19 浓度呈正相关。 此外, 角蛋白 19 的这一作用与 PTEN 和 MAPK 通路有关。 结论 细胞角 蛋白 19 能够明显提高 B-CPAP 增殖、 侵袭以及转移的能力, 同时角蛋白 19 的这一作用与 PTEN 和 MAPK 通 路有关。     

Novel monoclonal antibodies defining epitope of human cytokeratin 18 molecule

Two monoclonal antibodies, DA7 and DC10, were obtained from fusions of mouse myeloma cells with splenic lymphocytes from mice immunized with human breast cancer cells of PMC 42 line. The indirect immunofluorescence studies performed on established tumor cell lines together with immunoperoxidase staining of normal human tissues showed that the components reacting with the antibodies were cytokeratins. Positive reaction was noted in all epithelia derived cultured cells and in all simple epithelial tissues known to express keratin 18. Immunoblotting performed on various cytoskeletal preparations demonstrated strong staining of a single band with a mobility corresponding to that of cytokeratin 18 (45 kD). The negative immunoperoxidase reaction found in different epithelial tissues of seven animal species suggests that both antibodies are specific for human keratin 18. It was shown that DA7 and DC10 antibodies exhibited strong reaction in paraffin embedded tissues fixed in either methacarn or standard formalin. These characteristics predetermine both antibodies as suitable reagents for the specialized histopathological work.     

抗人肝癌单克隆抗体HL2的制备及鉴定

用肝癌细胞株QGY-7703、BEL-7402、SMMC-7721以贯序法免疫BALB/c小鼠,获得一株分泌肝癌单抗的杂交窟株HL_2,其染色体数目为97—105。单抗HL_2系IgG_2b亚类。吸收实验及阻断实验证明HL_2抗原与CEA、AFP、HBsAg、HBcAg及HBeAg无免疫同源性。ABC法和ELISA法说明单抗HL_2与四株肝癌细胞、六例肝癌组织均呈明确阳性反应,除与SGC-7901、H_(128)、K_(562)、Hela细胞株及部分胚胎肝脏,胚胎结肠有较弱性反应外,与肝癌旁组织、正常肝脏、其它肿瘤组织和细胞株、二种正常人细胞及其它胚胎组织无反应。显示了单抗HL_2的特异性较好、阳性率较高,是一个有应用前景的单抗。     

Use of monoclonal antibodies reactive with secretory epithelial cells for the immunocytochemical identification of plasma cells

Six monoclonal antibodies (McAbs) were identified as plasma cell-reactive when screened on sections of human tonsil. They were all produced following immunisation of mice with cells of a human plasmacytoid line. Three of the antibodies also stained the cytoplasm (but not the surface) of blood B cells and were unreactive with other leucocytes; one McAb showed broad lymphocyte reactivity and two were completely unreactive with blood leucocytes; on testing with a panel of cell lines specificity for the plasmacytoid line was demonstrated by three of the McAbs. In spite of the marked restriction shown by the reactivity of these antibodies in tests on cells of haemopoietic origin, tests on other human tissues - including thyroid and pancreas - showed that a related antigen was present in the cytoplasm of secretory epithelial cells. The overall patterns of reactivity of the individual McAbs on various tissues and blood lymphocytes were different. Comparisons were made with the established McAb OKT10, which binds to plasma cells, early stem cells and activated lymphocytes; its binding to plasma cells was confirmed and it was shown that it did not stain secretory epithelia. The potent reactions obtained with the new McAbs suggest that antibodies to antigens associated with epithelial cell secretory apparatus provide potentially useful reagents for studying plasma cells.     

Use of monoclonal antibodies to keratin 7 in the differential diagnosis of adenocarcinomas.

Monoclonal antibodies (MAbs) to specific keratin subtypes were prepared and characterized by immunoblotting and immunohistochemical assays on human cell cultures and normal and malignant human tissues. Chain-specific MAbs to keratin 7 (RCK 105, OV-TL 12/30) and keratin 18 (RGE 53, RCK 106, CK18-2), as well as broadly cross-reacting keratin MAbs (RCK 102, OV-TL 12/5) could be shown to react with different types of human epithelial tissues and were therefore tested for their usefulness in the differential diagnosis of carcinomas. The two broad-spectrum antibodies stained virtually all of the more than 350 carcinomas tested, especially when combined, and distinguished them from most nonepithelial tumors. The keratin 18 MAbs distinguished adenocarcinomas (which are keratin 18 positive) from most squamous cell carcinomas (which are generally keratin 18 negative). The MAbs to keratin 7 could be shown to recognize specific subtypes of adenocarcinoma and could, for example, distinguish between ovarian carcinomas (keratin 7 positive) and carcinomas of the gastrointestinal tract (keratin 7 negative), or between transitional cell carcinomas (keratin 7 positive) and prostate cancer (keratin 7 negative). In general, malignancies showed the expected keratin reactivity pattern as concluded from the keratin pattern of its cell of origin or its type of differentiation. The use of an extended series of malignancies did, however, also illustrate that exceptions to this rule exist. For example, certain antibodies to keratin 18 stained tumor areas in squamous cell carcinomas of the lung. Also a certain percentage of tumors, which generally showed no keratin 7 expression, were positive with RCK 105 or OV-TL 12/30. On the other hand, a certain percentage of tumors, which were generally positive for keratin 7, did not show a staining reaction with these MAbs. Furthermore subtle differences between reactivity patterns of different MAbs recognizing the same keratin protein were observed, both in the normal and malignant human tissues, indicating that specific keratin epitopes may be masked in certain tissues and that unmasking of such epitopes can occur with malignant progression. This phenomenon may be of some use in a further subtyping of carcinomas, especially those of the gastrointestinal tract. Despite these exceptional staining patterns, the keratin MAbs described above have proved to be useful tools in the characterization of epithelial tumors in routine histopathology and cytopathology, in which they add to a more refined diagnosis of (adeno)carcinomas.     

Monoclonal antibodies specific for subsets of epidermal keratins: biochemical and immunocytochemical characterization--applications in pathology and cell culture

Keratin composition has been widely used as a biochemical marker of differentiation in normal epithelia, cell culture systems and tumours of epithelial tissues. We have been developing a model system for the study of human squamous epithelial cell differentiation, and among a panel of monoclonal antibodies we have generated for analysing this system are two antibodies recognizing subsets of epidermal keratins. The two antibodies, designated LICR-LON-16a and LICR-LON-29b, were raised to the human squamous carcinoma cell line LICR-LON-HN-5, and we describe here their biochemical and immunocytochemical characterization. Antibody 16a reacts with only epidermal basal cells in normal human skin and shows specificity for the 45 and 46 kdalton keratins. Antibody 29b stains all living layers of the epidermis, and reacts with a broad range of ketain polypeptides, (45-56 kdaltons) in immunoblotting analyses. We have investigated the alterations of cellular staining that occur in chronic hyperproliferative skin diseases and carcinomas and compared this with the staining of multilayered cultures of normal keratinocytes and the HN-5 cell line. We show that in squamous cell carcinomas and in HN-5 cell xenografts 16a and 29b stain only the well-differentiated cell types. Furthermore we found that the basal cell specificity of 16a was lost in all of the hyperproliferative skin lesions examined including psoriasis and eczema. This transition to suprabasal staining pattern was also seen in the cultures of normal keratinocytes and HN-5 cells. We conclude that aberrant keratin synthesis or abnormal post-translational processing of keratins associated with an increased rate of cell turnover could account for the altered expression of the epitope recognized by antibody 16a.     

Differential diagnosis of gastrointestinal carcinomas by using monoclonal antibodies specific for individual keratin polypeptides

Monoclonal antibodies which recognize particular keratin polypeptides have been used to analyze normal human tissues including pancreas, stomach, colon, gall bladder, and liver as well as tumors of the gastrointestinal tract by immunohistological techniques. Broad specificity (lu5), ker 8 (Troma 1) and ker 18 (CK2) antibodies were positive while a ker 14 specific antibody (CKB1) was negative on all specimens tested. Differential staining patterns were seen with a ker 7 (CK7) and a ker 19 (KA4) antibody. Both antibodies stained gall bladder epithelium, pancreatic ducts but not acinar cells, as well as pancreatic ductal adenocarcinomas. KA4 but not the CK7 antibody stained adenocarcinomas of the stomach and large bowel. Both antibodies stained bile ducts and cholangiocellular carcinoma of the liver but did not stain hepatocytes or hepatocellular carcinomas. The results with keratin monoclonal antibodies compare well with those obtained by others using two dimensional gel electrophoresis and they further support the idea that monoclonal antibodies specific for particular keratin polypeptides will find applications in routine pathological diagnosis.     

Monoclonal antibodies to the human mammary gland

Summary Mouse monoclonal antibodies have been raised to the human milk fat globule membrane. The distribution of the antigens detected by four of the antibodies has been examined in formalin-fixed, paraffin-embedded human tissues by light microscopic immunocytochemistry. The four antibodies stain lactating breast and normal resting breast. Two exclusively stain the luminal membranes of breast epithelial cells. A third antibody stains in addition the lateral membranes of duct epithelial cells. The fourth antibody stains both epithelial and myoepithelial cells. None of the antibodies is breast specific, nor do they stain every epithelial cell within the breast. Instead, each antibody reveals a complex and heterogeneous distribution of staining throughout the normal tissues. Within the breast, the staining by a given antibody is usually segmental and conforms to secretory units and their associated ducts. Similarly heterogeneous patterns of staining are also observed in the extramammary normal tissues. Despite the apparent morphological identity between breast epithelial cells when examined by conventional light microscopy, the hitherto unrecognised functional heterogeneity, which has been revealed by the monoclonal antibodies could have importance in understanding the biology of the normal breast and the pathology of breast cancer.     

Mesenchymal specificity of three new monoclonal antibodies generated to the osteosarcoma cell line 4444T

We have generated three murine monoclonal antibodies to the new human osteosarcoma cell line 4444T. Analysis of their binding patterns to tumor cell lines, normal human tissues, and surgical tumor specimens indicates that the antibodies recognize a subset of human sarcomas and stromal tissues but fail to react with carcinomas or normal human epithelial tissue. These mesenchyme-specific monoclonal antibodies bind to antigens in the extracellular matrix. One antibody is specific in its binding to the muscularis arteriosus. We expect these antibodies to aid in the identification of sarcomas and to extend our knowledge of the components of the extracellular matrix and its interaction with tumors.     

Keratin and vimentin distribution patterns in the epithelial structures of the cannie anal region

The intermediate filament labeling pattern of the epithelial structures of the canine anal region was studied with different polypeptide specific keratin monoclonal antibodies (MoAbs) and with a monoclonal and polyclonal vimentin antibody. The epithelial structures in this region could be discriminated and characterized by differences in their keratin staining pattern. The basal cells in the different epithelial structures showed a similar staining pattern characterized by reactivity with MoAbs staining keratins 5, 8, 14, and 17. Columnar epithelial cells showed a completely different phenotype mostly characterized by reactivity with MoAbs staining keratins 7, 5, 8, 18, and 19. A restricted number of differentiated perianal gland cells showed perinuclear vimentin staining. Myoepithelial cells did not stain for vimentin, but, as other basal cells, were positive for MoAbs staining keratins 5, 8, 14, and 17.© Willey-Liss, Inc.     

Keratin and vimentin distribution patterns in the epithelial structures of the canine anal region.

The intermediate filament labeling pattern of the epithelial structures of the canine anal region was studied with different polypeptide specific keratin monoclonal antibodies (MoAbs) and with a monoclonal and polyclonal vimentin antibody. The epithelial structures in this region could be discriminated and characterized by differences in their keratin staining pattern. The basal cells in the different epithelial structures showed a similar staining pattern characterized by reactivity with MoAbs staining keratins 5, 8, 14, and 17. Columnar epithelial cells showed a completely different phenotype mostly characterized by reactivity with MoAbs staining keratins 7, 5, 8, 18, and 19. A restricted number of differentiated perianal gland cells showed perinuclear vimentin staining. Myoepithelial cells did not stain for vimentin, but, as other basal cells, were positive for MoAbs staining keratins 5, 8, 14, and 17.