Beta-tubulin epitope expression in normal and malignant epithelial cells.

The expression of a unique beta-tubulin isoform (class III) was monitored in squamous cell carcinoma (SCC) and normal epithelial cells using a monoclonal tubulin antibody called TuJ1. Whole tissue homogenates of SCC, normal tissue, SCC grown in nude mice, and SCC cultured cells were examined using sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot. TuJ1 antibody localization was performed using peroxidase immunostaining on paraffin sections of SCC, normal tissue, nude mouse SCC, and immunofluorescent microscopy of SCC cultured cells. The malignant tissues examined stained positive with TuJ1 and a general beta-tubulin antibody, whereas the normal tissues stained positively only for the general beta-tubulin antibody. TuJ1 epitope expression may be a useful marker for SCCs and may assist in understanding differences between normal and malignant squamous cells.     

Overexpression of the A9 antigen/alpha 6 beta 4 integrin in head and neck cancer.

A tumor-associated antigen detected by monoclonal antibody UM-A9 raised against a cultured cell line from a patient with an aggressive SCC of the oral cavity has been defined. The A9 antigen is abnormally expressed in squamous cancers, with loss of basal polarization and increased intensity of expression distinguishing malignant from normal cells. A minority of cultured SCC cell lines and about one third of fresh tumors exhibit polarized A9 expression. The increased intensity and loss of polarized expression of A9 antigen in recurrent and metastatic tumor cell lines when compared with primary or early tumor cell lines from the same patients indicated an association of altered expression with tumor progression. When A9 expression was evaluated in frozen tumor sections, three patterns of expression representing increasing intensity and loss of polarization were observed. Patients whose tumors exhibited the most intense A9 antigen expression had a higher rate of early relapse than patients whose tumors exhibited low intensity and polar expression. Loss of blood group antigen expression was also associated with poor prognosis, and together high A9 antigen expression and loss of blood group defined a group of patients at high risk of early relapse. The A9 antigen is immunologically and biochemically identical to the alpha 6 beta 4 integrin. The association of high expression of the A9/alpha 6 beta 4 integrin as a prognostic factor is supported by similar findings with a mouse model system. The mouse tumor-associated antigen, TSP-180, which is also an alpha 6 beta 4 integrin, distinguishes highly metastatic tumor cells from nonmetastatic variants of the same tumor line. In SCC, the alpha 6 beta 4 integrin contributes to attachment to laminin since anti-alpha 6 subunit specific antibody blocks cell attachment and only the beta 4 subunit is found in association with the alpha 6 subunit in these cells. Similar findings were obtained in colon carcinomas. Antibodies and peptides that block laminin attachment may lead to the development of antimetastatic agents for squamous carcinomas. The beta 4 subunit is unique from other integrins in that it has an unusually long cytoplasmic domain, the function of which is not known. The beta 4 subunit is heavily phosphorylated under conditions that favor anchoring and terminal differentiation in normal keratinocytes. Paradoxically the beta 4 subunit is also heavily phosphorylated in tumor cells, which are highly migratory and immortalized.(ABSTRACT TRUNCATED AT 400 WORDS)     

Immunohistochemical identification of squamous carcinoma of the head and neck with monoclonal antibody SQM1

Frozen tissue sections of biopsies from head and neck squamous cancer lesions were examined for immunohistochemical staining with a recently developed monoclonal antibody, designated as SQM1 antibody and directed against the surface membrane of squamous carcinoma cells. SQM1 antibody stained selectively squamous carcinoma, while normal mucosa and cells of the stroma were non-reactive. Positive staining of tumor was found in 33/35 specimens obtained from several major sites of the head and neck area and was observed in primary manifestations and lymph node metastases as well as in recurrences. The most consistent reactivity was seen with carcinomas of the tongue. Well differentiated squamous carcinomas contained a higher proportion of SQM1 positive tumor cells than poorly differentiated carcinomas. We suggest that the SQM1 antibody may aid in the immunohistochemical identification of squamous carcinoma of the head and neck area.     

Monoclonal antibodies (Mabs) in head and neck cancer

The development of monoclonal antibodies (Mabs) represents a major contribution of immunology to the study of head and neck cancer. The interest of the otolaryngologist in this new field is quite recent, but our knowledge of cancer and other immunoproliferative disorders will exponentially expand with the characterization of tumor-associated antigens of squamous cell carcinomas. The author reviews briefly the technique for producing monoclonal antibodies and presents an overview of the application of Mabs to clinical medicine, oncology and, more specifically, head and neck cancer. Very few contributions have been made in head and neck cancer, but the field of Mabs will significantly alter our diagnostic and therapeutic armamentarium.     

免疫组化和连续切片在检测头颈鳞癌颈淋巴结微转移中的意义

目的〓〖HTK〗探讨应用连续切片EnvisionTM法检测角蛋白表达对判断头颈鳞状细胞癌淋巴结微转移的效果及微转移对肿瘤预后的影响。〖HTW〗方法〓〖HTK〗术中淋巴结标本行快速EnvisionTM法检测角蛋白CKAE1/AE3,与常规冰冻病理对淋巴结微转移检出率比较。对14例下咽癌、声门上型喉癌淋巴结标本连续病理切片,比较常规病理检测与EnvisionTM法对淋巴结微转移检出率。〖HTW〗结果〓〖HTK〗快速EnvisionTM法可检出2粒常规冰冻病理不能检出的淋巴结微转移,连续切片应用EnvisionTM法可检出5粒常规病理阴性的淋巴结（5/216）存在微转移灶。〖HTW〗结论〓〖HTK〗EnvisionTM法检测角蛋白CKAE1/AE3表达可快速判断头颈部鳞状细胞癌淋巴结微转移,存在多粒淋巴结微转移的患者预后可能较差。     

喉鳞癌组织两种P糖蛋白单克隆抗体的检测

为了解喉鳞癌的多药耐药性，以期提高综合治疗中化疗的效果，采用免疫组化技术对未经任何治疗的喉鳞癌患者在手术中取出的新鲜肿瘤组织进行了Ｐ糖蛋白（ＰＧＰ）单克隆抗体ＪＳＢ１及其单克期抗体Ｃ２１９的检测。对比了肿瘤组织分化程度、肿瘤大小及有无淋巴结转移与ＰＧＰ表达阳性之间的关系。结果发现，单抗ＰＧＰ ＪＳＢ１的阳性率为６５．２％（ｎ＝２３），而单抗Ｃ２１９检测的６例ＰＧＰ均为阴性，与分化无关，与肿瘤大小、     

Monoklonale Antikörper gegen Merkmale der Zellmembran von Larynxkarzinomzellen

Summary Humoral and cellular immune responses to laryngeal carcinomas suggest the presence of tumor associated antigens on the surface of larynx carcinoma cells. Production of antiserums against specific antigens on human tumors or tissues is complicated by the concomitant production of antibodies that react with all human cells. To circumvent this problem monovalent antibodies are needed.After isolation of larynx carcinoma cells by cell culture techniques outer cell membranes were prepared and used to immunize mice. From these mice the antibody producing spleen cells were isolated and fused with myeloma cells resulting in antibody secreting hybrid cells. Hybrid cells were cloned thus forming monoclonal antibodies. Monoclonal antibodies are a useful tool for detection of tumor specific antigens and differentiation antigens. They represent an approach toward identifying and isolating cell surface components.

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Ausführliche Publikation anderweitig vorgesehen     

Tumor-Associated glycoprotein expression in salivary gland mucoepidermoid carcinomas: An immunohistochemical study using the monoclonal antibody B72.3

The monoclonal antibody B72.3 recognizes a mucin-like glycoprotein which is expressed in a variety of human malignancies including adenocarcinomas of the breast, colon, lung, endometrium, pancreas, and ovary. The antibody, an immunoglobulin G (IgG1) subclass monoclonal antibody, was generated from membrane-enriched fractions of metastatic mammary carcinoma. Because the antigen detected by B72.3 is expressed in a broad spectrum of human neoplasms, B72.3 is a potentially useful marker for glandular neoplasms. Despite its potential usefulness, there is only one previous comprehensive study which has examined the distribution of the tumor-associated glycoprotein (TAG-72), as detected by B72.3, in salivary neoplasms. We examined 21 mucoepidermoid carcinomas with the antibody. Twenty (95%) of the 21 carcinomas stained positively. The only carcinoma that did not stain was initially thought to be a high-grade mucoepidermoid carcinoma based on immunohistochemical staining. Further review, however, indicated that this lesion was most likely a pure squamous cell carcinoma, and the absence of TAG-72 antigen expression was helpful in this regard. Among those neoplasms which stained with B72.3, the strongest staining was seen in the low-grade carcinomas. These results suggest that B72.3 may be a useful marker for glandular differentiation in mucoepidermoid carcinomas.     

鼻腔鼻窦内翻性乳头状瘤中Survivin及Bcl-2的表达

目的：探讨细胞凋亡抑制蛋白Survivin在鼻腔鼻窦内翻性乳头状瘤中的表达及其与Bcl-2表达的相关性。方法：采用免疫组织化学Envision二步法检测30例鼻腔鼻窦内翻性乳头状瘤,10例鼻腔鳞状细胞癌及10例正常下鼻甲黏膜标本中Survivin和Bcl-2的表达。结果：鼻腔鼻窦内翻性乳头状瘤和鼻腔鳞状细胞癌组织中Survivin的表达分别为73.3%和80.0%,均显著高于正常下鼻甲黏膜组织（均P〈0.01）。鼻腔鳞状细胞癌组织中Bcl-2的表达率为90.0%,显著高于正常下鼻甲黏膜组织的表达率（20.0%）（P〈0.01）。鼻腔鼻窦内翻性乳头状瘤组织中Bcl-2的表达率为46.7%,高于正常下鼻甲黏膜,但两者之间的差异无统计学意义（P〉0.05）。Survivin表达与Bcl-2表达呈正相关（P〈0.01）。结论：Survivin在鼻腔鼻窦内翻性乳头状瘤和鼻腔鳞状细胞癌的发生、发展中起重要作用,可能成为鼻腔鼻窦内翻性乳头状瘤和鼻腔鳞状细胞癌基因治疗的靶点。Survivin可能与Bcl-2在鼻腔内翻性乳头状瘤的发展中起协同作用。     

Development of monoclonal antibodies with specificity to oral squamous cell carcinoma

A panel of five monoclonal antibodies have been produced that show binding when directed against human oral head and neck squamous cell carcinomas. These monoclonal antibodies are derived from antibody-secreting cells obtained using cell hybridization techniques. The activity of the antibody was tested in vitro against oral head and neck squamous cell carcinoma (n = 10). Control groups included normal oral mucosa (n = 7), hyperkeratosis (n = 5), mild dysplasia (n = 4), and severe dysplasia (n = 3). Results using immunoperoxidase staining confirmed the binding of the five monoclonal antibodies to human head and neck squamous cell carcinoma cells, with predominant reactivity (50% to 90%) in the majority of the specimens, compared with negative (0%) to weak (less than 5%) or focal (5% to less than 50%) reactivity in the control specimens.     

Interest in frozen section examination of margins and lymph nodes in laryngeal surgery

One hundred and one patients presenting with a squamous cell carcinoma of the larynx underwent surgery in our department between January 1980 and May 1985. In most of these patients, nodes were removed from the main lymphatic drainage pathways and subjected to immediate frozen section examination. The results from frozen section examination of the nodes were then compared with those from the surgical specimens of cervical neck dissections performed on the patients according to the classic rules. In addition, margin resections were made and examined by frozen section after removal of the tumour. In the event of a positive finding, these resections were continued until healthy tissue was reached, the specimens being examined in addition by classic methods. Immediate frozen sections enable the margins of the resection to be verified correctly. In our series we were brought to extend the limits of resection in 10 cases out of 68 (15 per cent). It can also be seen that the accuracy of the pathologist's reading of the frozen sections is satisfactory. The overall level of error is three out of 68 (4.5 per cent). All the errors correspond to false negatives. The aim of avoiding neck dissections in the presence of N0, thanks to nodal selection with frozen section, is not attained. We find a 6/61 rate of false negatives for N0-N1 (10 per cent) when we compare the frozen sections of the selected nodes and the neck dissections. This is due to the fact that the surgeon may be led astray by a reactive hyperplastic node whilst other less inflammatory neighbouring nodes are in fact the site of metastasis.     

喉癌相关抗原免疫电镜的定位研究

目的研究喉癌混合单克隆抗体相关抗原的定位,为喉癌的早期诊断及导向药物治疗奠定理论基础。方法采用三株喉癌单克隆抗体ＬＣ９、ＬＣ１１、ＬＣ１２混合对９０例病理证实的喉癌组织切片（病理Ⅰ级４９例,Ⅱ级４１例）、１４例喉癌前病变及１０例正常喉粘膜进行免疫组化染色（ＡＢＣ法）光镜观察,同时采用ＬＳＡＢ法对９例新鲜喉癌组织细胞行免疫过氧化物酶标记电镜包埋前染色,对混合单克隆抗体相应抗原进行超微结构的定位观察。结果喉癌组织中喉癌相关抗原检出率（９７．７％）与正常喉粘膜相比明显增高（Ｐ＜０．０１）;混合单克隆抗体相应抗原主要定位于细胞膜性结构,即细胞膜、核膜、线粒体膜和内质网膜。结论混合喉癌单克隆抗体具有良好的特异性,带有药物的单克隆抗体可通过膜性结构来攻击癌细胞,达到治疗作用。混合单克隆抗体检测的相关抗原均分布于细胞膜和／或细胞浆中,细胞核无着色。     

Oncofetal antigen expression in head and neck carcinoma.

Oncofetal antigens (OFA) are proteins expressed during periods of embryonal/fetal development and on malignant cells. Previous investigations detected the presence of phase-specific 44- and 200-kd OFA, using a monoclonal antibody (MoAb 115). This study will determine if primary head and neck squamous cell carcinomas express the 44-kd OFA, and will assess the possible role of OFA in oncogenesis. Fifteen primary human head and neck squamous cell carcinomas (HNSCC) and six HNSCC cell lines were tested for OFA expression. Thirteen primary cancers and three cell lines demonstrated various degrees of OFA positivity. Oncofetal antigens are proposed to represent a proto-oncogene active during fetal development and malignant growth in head and neck squamous cell carcinomas.     

Studies on localization of laryngeal carcinoma associated antigen by immunoelectromicroscopy]

OBJECTIVE: To study the localization of laryngeal carcinoma associated antigen (LCAA) in the carcinoma tissue. METHODS: Ninety cases of laryngeal carcinoma and 14 cases of laryngeal precancerous lesion and 10 cases of normal laryngeal mucosa were detected with three strains of monoclonal antibodies LC9, LC11, LC12 by immunochemistry. The positive sections of laryngeal carcinoma were observed under light microscope and electromicroscopy. RESULTS: The positive rates of LCAA were dramatically higher than that in normal epithelial and precancerous tissues (P < 0.01). The results showed that the mixed monoclonal antibody had tissue specificity. The MLC associated antigens only distributed in cell membranes and/or cytoplasm. No cell nucleus was stained. CONCLUSION: The LCAA is mainly located in cell membranous structure. This study may provide morphological basis for immunoimaging diagnosis and targeting chemotherapy by application of laryngeal carcinoma McAb.     

A simple objective evaluation and grading for facial paralysis outcomes

Matrix metalloproteinase (MMP)-2 and -9 degrade type IV collagen, which is one of the major components of the basement membrane in normal tissue and expressed in the surroundings of the cancer nest in squamous cell carinoma. The degeneration of type IV collagen is an essential step in the metastasis to lymph nodes and distant organs. In this study, we examined MMP-2 and -9 levels of cancer tissue and serum obtained from patients with head and neck squamous cell carcinoma (HNSCC) in order to evaluate the relationship between the clinicopathologic features and MMPs. We examined the production of MMP-2 and -9 in cancer tissue homogenates of 73 patients who had HNSCC and the serum MMP levels of 16 patients with HNSCC and 8 healthy volunteers. We also studied the localization of MMP-2 in the carcinoma using an immunohistochemical approach. The concentrations of MMP-2 and -9 in the tissue homogenates and serum were measured by means of a sandwich enzyme immunoassay using a monoclonal antibody. Immunohistochemical analyses were performed with monoclonal antibody to MMP-2. The concentration of MMP-2 in the tumor tissue homogenates was unrelated to tumor size, but that in patients with lymph node metastases was significantly higher than in those without lymph node metastases. The concentration of MMP-9 was unrelated to lymph node metastasis and tumor size. The levels of both MMP-2 and -9 in serum were unrelated to lymph node metastasis. Immunohistochemistry indicated that MMP-2 was mainly expressed in cancer cells. Because MMP-2 degrades type IV collagen, the level of MMP-2 in carcinomas may be a useful indicator of the degree of invasion and metastasis.     

Enhanced production of matrix metalloproteinase-2 in human head and neck carcinomas is correlated with lymph node metastasis

Matrix metalloproteinase (MMP)-2 and -9 degrade type IV collagen, which is one of the major components of the basement membrane in normal tissue and expressed in the surroundings of the cancer nest in squamous cell carcinoma. The degeneration of type IV collagen is an essential step in the metastasis to lymph nodes and distant organs. In this study, we examined MMP-2 and -9 levels of cancer tissue and serum obtained from patients with head and neck squamous cell carcinoma (HNSCC) in order to evaluate the relationship between the clinicopathologic features and MMPs. We examined the production of MMP-2 and -9 in cancer tissue homogenates of 73 patients who had HNSCC and the serum MMP levels of 16 patients with HNSCC and 8 healthy volunteers. We also studied the localization of MMP-2 in the carcinoma using an immunohistochemical approach. The concentrations of MMP-2 and -9 in the tissue homogenates and serum were measured by means of a sandwich enzyme immunoassay using a monoclonal antibody. Immunohistochemical analyses were performed with monoclonal antibody to MMP-2. The concentration of MMP-2 in the tumor tissue homogenates was unrelated to tumor size, but that in patients with lymph node metastases was significantly higher than in those without lymph node metastases. The concentration of MMP-9 was unrelated to lymph node metastasis and tumor size. The levels of both MMP-2 and -9 in serum were unrelated to lymph node metastasis. Immunohistochemistry indicated that MMP-2 was mainly expressed in cancer cells. Because MMP-2 degrades type IV collagen, the level of MMP-2 in carcinomas may be a useful indicator of the degree of invasion and metastasis.     

Basaloid squamous cell carcinoma of the maxilla: a case report and immunohistochemical analysis

Basaloid squamous cell carcinoma (BSCC) is a recently recognized high-grade tumor with a propensity for nodal as well as systemic metastasis and can arise from different anatomic locations. The differential diagnosis includes adenoid cystic carcinoma, small cell neuroendocrine carcinoma and squamous cell carcinoma. Monoclonal antibodies reactive with cytokeratin (34betaE12, AE3, pancytokeratin), as well as other cellular antigens (vimentin [VIM]; synaptophysin [SYNF]; chromogranin A [ChA]; neuron-specific enolase [NSE]; S-100, desmin, smooth-muscle actin [SMA]), were used in an immunoperoxidase method with paraffin-embedded tissue to phenotypically characterize a case with features of BSCC arising in the maxillary sinus. Neoplastic cells reacted with the high-molecular-weight cytokeratin antibody 34betaE12, as well as with other antikeratin antibodies, but failed to react with the antibodies VIM, desmin and SMA and showed variable immunoreactivity for NSE, SYNF and S-100. The staining pattern for NSE was diffuse and intense and reactivity for ChA was inconsistent.     

Monoclonal antibodies to inner ear antigens: I. Antigens expressed by supporting cells of the guinea pig cochlea

Murine monoclonal antibodies against guinea pig cochlear epithelium were generated with the goal of identifying cochlea-specific antigens and elucidating their function. To compensate for the limited amount of cochlear tissue, intrasplenic immunization was used. Hybridoma supernatants were screened by ELISA for antibody production and for binding to homogenates from cochlea, liver, lung, kidney and brain. Hybrids producing antibody to cochlea were subcloned and tested immunocytochemically against frozen sections and surface preparations of paraformaldehyde-fixed cochlear tissue. KHRI-1, a low titer IgM antibody stained only Hensen cells. KHRI-2, also an IgM antibody, stained tectorial membrane, cells of the spiral limbus, cells bordering the space of Nuel, Hensen cells and the root cells of the spiral prominence. KHRI-3, an IgG1 antibody, stained the phalangeal processes of outer pillar cells and the apical portion of phalangeal processes of Deiters' cells in a distinctive wine goblet pattern on surface preparations. KHRI-3 antibody also reacted with peripheral nerves and pia mater of brain in unfixed frozen sections but the antigenic site was not stable to fixation in contrast to the epitope detected in the cochlea. In Western blots of detergent extracts from cochlea KHRI-3 stained a broad tissue-specific band of Mr 70-75 kDa; a narrower band of Mr 68-70 kDa was identified by KHRI-3 in extracts of tongue and brain. KHRI-1 and KHRI-2 did not detect any proteins in Western blots. The monoclonal antibodies KHRI-1, -2, and -3 which define epitopes expressed by discrete populations of supporting cells in the inner ear should be useful in characterizing the nature and function of cellular structures in the cochlea.     

Expression of KITENIN and its association with tumor progression in oral squamous cell carcinoma

Objective

KAI1 COOH-terminal interacting tetraspanin (KITENIN) contributes to tumor invasion and metastasis in various cancers. The aim of this study was to investigate expression of KITENIN in patients with oral cavity squamous cell carcinoma (SCC) and to determine whether KITENIN affects tumor cell behavior in oral cavity SCC cell line.

Methods

Western blotting and immunohistochemistry was used to assess alteration of KITENIN expression in human oral cavity SCC and normal oral cavity mucosa. To evaluate the impact of KITENIN knockdown, the cell invasion assay and cell migration assay using small-interfering RNA were performed.

Results

KITENIN protein expression was significantly increased in human oral cavity SCC tissues than in normal oral cavity mucosa by Western blotting. KITENIN immunoreactivity was strongly identified in human oral cavity SCC relative to adjacent normal tissue. Knockdown of KITENIN resulted in significantly reduced cell invasion in human oral cavity SCC cells (p = 0.001). Cell migration showed a marked decrease in KITENIN knockdown oral cavity SCC cells compared to the negative control oral cavity SCC cells (p = 0.01).

Conclusion

KITENIN is associated with tumor invasiveness and metastasis in human oral cavity SCC.     

Fluorescent in situ hybridizaton evaluation of p53 gene deletions at a tumor interface of lingual carcinoma

OBJECTIVE/HYPOTHESIS: To evaluate the ability of fluorescent in situ hybridization (FISH) to detect malignant cells missed by standard histological assessment at an interface between malignant and normal tissue in lingual squamous cell carcinoma (SCC) and to correlate findings of FISH assessment with patients' clinical stages. STUDY DESIGN: Retrospective assessment of archival tissue from 31 patients with lingual SCC treated at University of Wisconsin Hospital and Clinics in Madison. METHODS: An assay combining standard histological and FISH techniques was used to assess a tumor interface tissue section and allow identification of each tumor's ploidy characteristics and p53 gene deletions and the presence or absence of malignant cells within tissue viewed as "normal" on histological review. RESULTS: Forty-five percent of tumors (14 of 31) demonstrated ploidy changes and 84% (26 of 31) showed p53 deletions. Of these 26 tumors with p53 deletions, 14 were found to have "microfoci" with p53 deletions within tissue that appeared normal on histological examination. These microfoci were found in 75% of late-stage tumors and in only 35% of early-stage tumors. CONCLUSIONS: FISH allowed identification of malignant cells in tissue viewed as normal on standard histological assessment, and this finding occurred more frequently in late-stage tumors.