Ber EP4 and epithelial membrane antigen aid distinction of basal cell, squamous cell and basosquamous carcinomas of the skin

AIMS: Seventy-five skin tumours were studied to investigate the value of immunohistochemistry in differentiating basal cell, squamous cell and basosquamous carcinomas of the skin. METHODS AND RESULTS: Archived paraffin-embedded tissue samples of basal cell carcinomas (n = 39), squamous cell carcinomas (n = 23) and basosquamous carcinomas (n = 13) were stained immunohistochemically using a panel of antibodies. All of the basal cell carcinomas stained positively for Ber EP4, in contrast to the group of squamous cell carcinomas, that showed no staining. Basosquamous carcinomas all showed at least some areas of Ber EP4 positivity. None of the basal cell carcinomas, but most of the squamous cell carcinomas (22 of 23) expressed epithelial membrane antigen (EMA). Only one of the basosquamous carcinomas expressed EMA positivity focally. CAM 5.2, carcinoembryonic antigen (CEA) and 34betaE12 antibodies lacked specificity in relation to the different tumour types. CONCLUSION: Distinction of basal and squamous cell carcinomas of the skin can be readily achieved with routine immunohistochemistry using Ber EP4 and EMA. Identification of basosquamous carcinoma is also facilitated with this method.     

High-level expression of CD109 is frequently detected in lung squamous cell carcinomas

CD109 is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein, which is a member of the alpha2-macroglobulin/C3, C4, C5 family of thioester-containing proteins. It has been reported that CD109 is expressed in a subset of hematopoietic cells, endothelial cells and several kinds of human tumors. Herein it is reported that the CD109 protein is preferentially expressed in lung squamous cell carcinomas compared with other types of lung carcinoma including adenocarcinomas, large cell carcinomas and small cell carcinomas. Immunohistochemical staining of surgically resected lung specimens using an anti-CD109 antibody detected CD109 expression in basal cells of bronchial and bronchiolar epithelia and myoepithelial cells of bronchial secretary glands, but not in bronchial and bronchiolar apical epithelial cells and alveolar epithelial cells. Furthermore, the CD109 immunoreactivity was observed in squamous cell carcinomas at a high frequency compared with other types of lung carcinoma. Although the detailed function of CD109 protein is unclear, these results suggest that CD109 expression may play a role in the development of lung squamous cell carcinoma.     

Lack of lymphangiogenesis during breast carcinogenesis

BACKGROUND: Recent evidence suggests that functional intratumorous lymph vessels may be absent from some human cancers. This could result from either the failure of tumours to induce lymphangiogenesis, or the collapse of lymph vessels, caused by high interstitial tumour pressure. METHODS: To differentiate between these two hypotheses, paraffin wax embedded clinical specimens from normal breast (n = 13), usual ductal hyperplasia (n = 11), ductal carcinoma in situ (n = 21), and invasive breast cancer (n = 40) were compared for lymphatic and blood vessel density by immunohistochemistry with antibodies to the lymphatic endothelial hyaluronan receptor (LYVE-1) and CD31, respectively. RESULTS: Lymph vessel density was lower than blood vessel density in normal breast tissue. Within breast lobuli, lymph vessels were absent. In premalignant lesions blood microvessel density increased, whereas no increase in lymph vessels could be seen intralesionally. In invasive cancers, lymph vessels were absent in all but a few cases, where probably some pre-existing lymph vessels remained, although blood microvessel density was once again increased. CONCLUSION: Unlike angiogenesis, lymphangiogenesis is absent during breast carcinogenesis. This, and not rising interstitial pressure caused by an increase in the size of lesions, explains the absence of intratumorous lymph vessels in invasive breast cancer.     

The human hematopoietic progenitor cell antigen (CD34) in vascular neoplasia.

The human hematopoietic progenitor cell antigen CD34 is synthesized and expressed by early normal hematopoietic progenitor cells and by many acute leukemias. Anti-CD34 antibodies also have been reported to stain blood vessels in tissue sections, and, more recently, CD34 mRNA has been detected in vascular endothelial cells. Therefore, the authors studied the diagnostic utility of immunohistochemical CD34 antigen detection in tumors of endothelial cell derivation and compared the results with stains for von Willebrand (vW) factor. A wide variety of epithelial and mesenchymal neoplasms also were examined to assess the specificity of CD34 for vascular neoplasia. Seven cases of angiosarcoma (seven of seven), five cases of Kaposi's sarcoma (five of five), and eight cases of epithelioid hemangioendothelioma (eight of eight) were moderately to strongly positive for CD34. This reactivity was equally intense in frozen sections, alcohol-fixed tissue, and formalin-fixed specimens. In many cases, the malignant endothelial cells stained more strongly than adjacent benign endothelium. Moreover, in most cases CD34 positivity was quantitatively and qualitatively stronger than staining for vW factor. Two cases of hemangiopericytoma (two of two) were CD34 positive but stained less intensely than the angiosarcomas, Kaposi's sarcomas, or hemangioendotheliomas. Five of six cases of hemangioma also stained positively for CD34; the nonreactive tumor in this group was the only one among 28 vascular neoplasms studied that was not reactive for CD34. In comparison, 9 of the 28 vascular tumors did not stain for vW factor. Three hundred fifty-seven tumors of nonvascular derivation also were examined for CD34 antigen expression. Focal light staining was seen in one pulmonary squamous cell carcinoma; moderate to intense staining was observed in half of the epithelioid sarcomas studied (8 of 16) and in a minority of leiomyosarcomas (3 of 22). These findings indicate that CD34 is a sensitive and relatively specific marker for neoplasms of vascular origin.     

Immunohistochemical and Immunoblot Analyses of ras-p21 Expression in Lung Carcinomas

Immunohistochemical and immunoblot analyses were performed on 49 cases of surgically resected primary lung carcinoma to examine the expression of ras oncogene product using monoclonal antibodies to ras -p21. Two different monoclonal antibodies, NCC RAS 001 and RAP 5 were used for immunohistochemical study. Cancer cells of 16 cases (33%) and 15 cases (31%) were strongly positive for NCC RAS 001 and RAP 5, respectively. The staining pattern of antibodies was heterogenous among cancer cells, even in the same case. Among various histologic type of lung cancers, squamous cell carcinomas and well-differentiated adenocarcinomas had a tendency to react more intensively than other histologic types of carcinoma. Immunoblot analysis using monoclonal antibody NCC RAS 004 revealed the presence of ras p21 not only in cancer tissues bus also in non-cancerous tissues in all cases analysed. In 13 cases (27%), cancer tissue expressed more than twise as much ras p21 as non cancerous tissues. Our study showed that the over expression of ras -p21 in lung carcinomas compared with non cancerous tissues was a relatively common phenomenon, especially in squamous cell carcinomas and adenocarcinomas. Acta Pathol Jpn 39: 643 647, 1989.     

Glypican-3 as a potential differential diagnosis marker for hepatocellular carcinoma: a tissue microarray-based study

The differential diagnosis between hepatocellular carcinoma (HCC) and benign hepatic lesions is still difficult and new biochemical markers for HCC are required. The aim of this study was to assess the differential diagnostic value of glypican-3 (GPC3) immunostaining in HCC patients. 147 cases of surgically excised HCC tissues, 94 cases from needle biopsies, and tissue microarrays were used for this study. The tissue microarrays contained 449 specimens including: 115 HCC, 25 intrahepatic cholangiocellular carcinoma, 29 lung adenocarcinoma, 23 squamous cell lung carcinoma, 53 ovary adenocarcinoma, 44 renal cell carcinoma, 30 prostate acinar adenocarcinoma, 42 breast carcinoma, 41 gastric carcinoma and 47 colorectal carcinoma. The immunolocalization of GPC3 was measured using immunohistochemical staining. Among 147 surgically excised HCC samples, 87.1% (128/147) were GPC3 positive. No GPC3 expression, however, was observed in paracarcinomatous and cirrhotic tissues. In needle biopsy tissues, GPC3 was positively expressed in 81.9% (77/94). Among tissue microassays, HCCs showed positive GPC3 expression in 55.7% (64/115), while 9.6% (5/52) of lung carcinoma and 5.7% (3/53) of ovary adenocarcinoma also were positively stained. The other tumor types showed negative GPC3 expression. In conclusion, our results show that GPC3 is specifically overexpressed in HCC tissue and may be regarded as a potential marker for differential diagnostic hepatocellular carcinoma.     

Study of the mechanism involved in angiogenesis and synovial cell proliferation in human synovial tissues of patients with rheumatoid arthritis using SCID mice

To examine whether synovial cell proliferation is due to angiogenesis, we studied the relationship between the inhibition of synovial cell proliferation and an angiogenesis inhibitor, TNP-470, in human synovial tissues. Human synovial tissues were implanted into the back of SCID mice (SCID-HuAg mice). Sixteen mice were divided into two groups of eight mice each: the untreated group (vehicle group) and the TNP-470-treated group that received a dose of 10 mg/kg body weight by subcutaneous injection. The number of blood vessels and synovial lining cells clearly increased in the vehicle group, but the number of synovial lining cells clearly decreased and the blood vessels were hardly detected in the TNP-470 group. Immunohistochemically, cells that stained positively for the anti-proliferating cell nuclear antigen (PCNA) mAb were abundant in synovial lining cells and endothelial cells in synovial tissues. Cells that stained positively for the anti-CD34 polyclonal antibody were abundant in the endothelial cells in the vehicle group, but these positively stained cells were hardly detected in the TNP-470 group. The PCNA positivity ratio in the vehicle group was 0.64 +/- 0.019, whereas that in the TNP-470 group was 0.199 +/- 0.007. The numbers of cells that stained positively for anti-CD34 polyclonal antibody were 242 +/- 13.4 in the vehicle group and 153 +/- 6.73 in the TNP-470 group per 10 microscopic fields. Cells that stained positively for anti-mouse CD31 mAb were mainly localized in the synovial lining, but invaded the subsynovial lining layer in human synovial tissues. On the other hand, cells that stained positively for anti-human CD31 mAb were mainly localized in the subsynovial lining layer. We found that endothelial cell proliferation is dependent on angiogenesis based on the result that angiogenesis and synovial cell proliferation were inhibited by treatment with TNP-470.     

人胎儿早中期肺免疫细胞的发育

目的：探讨人胎儿早中期肺免疫细胞的发生及发育特点。方法：采用免疫组化方法对胎儿肺免疫细胞的发生和发育进行观察。结果：肺间质中的白细胞共同抗原(leukocyte common antigen,LCA)阳性细胞和溶菌酶阳性细胞均在13周时出现;此后,两种阳性细胞数目有所增多,形态和分布上也有变化,且LCA阳性细胞数目较多。CD45-RO和CD20阳性细胞在28周开始出现于肺组织中。结论：人胎儿肺的固有免疫出现早,其中肺间质免疫细胞出现最早(13周),而支气管黏膜的稍迟(18周);适应性免疫则出现较迟(28周)。同时肺间质细胞在间质中免疫细胞功能成熟的过程中也起到促进和加速的作用。     

Lung cancer neovascularisation: Cellular and molecular interaction between endothelial and lung cancer cells

Background

Novel vascular-independent conduits have been observed in some cancers. These have been variously described as vasculogenic mimicry, mosaic vessel formation, vascular co-option and intratumour embryonic-like vasculogenesis. Despite lung cancer being the most common cancer worldwide, there is little information on its neovascularisation or the pathways involved.

Methods

An in vitro model involving co-cultures of microvascular lung endothelial cells and squamous or adenocarcinoma lung cancer cells was developed to assess their angiogenic interaction. Cells were incubated and examined by phase contrast microscopy and by immunocytochemistry in both mono- and co-cultures. Cultured cells and lung cancer tissue sections were assessed for new tumour vessel formation, expression of the endothelial marker CD31 and morphology.

Results

Lung tumour cells and endothelial cells interacted morphologically via pseudopodia and used alternative pathways to generate new vessels. Co-culturing microvascular endothelial and squamous carcinoma cells led to endothelial cells surrounding tumour cells and the tumour cells being incorporated into vessel walls. Co-culturing endothelial and adenocarcinoma cells resulted in cellular contact and the formation of tumour cell bridges around clusters of endothelial cells. These adencocarcinoma cells became strongly positive for CD31. Tumour tissue section studies supported the in vitro findings.

Conclusion

Lung carcinoma cells when co-cultured with lung endothelial cells modify their cellular and molecular features that encourage alternative means of providing blood supply. The mechanisms underpinning these non-angiogenic processes need to be further investigated and should be considered when anti-tumour therapeutic interventions are being considered.     

Matrix proteoglycans and remodelling of interstitial lung tissue in lymphangioleiomyomatosis

The interstitial lung disease lymphangioleiomyomatosis (LAM) is characterized by diffuse proliferation of smooth muscle cells (SMCs), which in many patients show TSC2 (tuberin) gene mutations, in addition to thickening of interstitial tissues, loss of alveoli, and the development of cystic spaces. While SMC proliferation is the defining feature of LAM, a significant proportion of LAM lung tissue consists of expanded interstitial connective tissue that is negative for smooth muscle actin and TSC2 mutations. The importance of this actin-negative interstitial tissue to the pathophysiology of LAM is not clear. The present study has determined the contribution of this interstitial tissue to LAM lung volume by morphometric analysis and has examined its cell and matrix proteoglycan composition by immunohistochemistry. Lung tissue from nine LAM patients and four control subjects was examined. LAM lung contained twice as much interstitial tissue as control lung (27% versus 13% of total lung volume), with SMCs accounting for less than 25% of the interstitial volume. Areas of interstitial tissue stained strongly for the matrix proteoglycans versican and biglycan. Decorin was prominent in association with collagen bundles. SMCs did not stain, or stained lightly, for proteoglycans. Versican and biglycan deposits were closely associated with actin-negative interstitial fibroblasts identified by prolyl 4-hydroxylase immuno-staining. Comparatively normal alveolar walls in LAM lung also stained strongly for versican and had a reduced elastin content. Thickened interstitial regions contained significant amounts of elastin (approximately 13% of interstitial volume) but with fibres in disorganized patterns. Elastic fibres were absent from areas that stained strongly for versican and biglycan. These areas also showed weak staining for elastin binding protein (EBP), consistent with proteoglycan-induced shedding of EBP and inhibition of elastic fibre formation. These findings point to a significant contribution from matrix proteoglycans to the expanded and remodelled interstitial lung tissue of LAM patients.     

Pathological study of carbohydrate antigens in human lung cancer with monoclonal antibodies]

The expressions of carbohydrate antigens were examined with panel of specific anti-carbohydrate monoclonal antibodies (MAbs) on human lung cancer tissues. The MAbs used were SH1, SH2, SNH3, AH6, CA3F4, TKH2, TKH6 and TKH5 which define Lex, dimeric Lex, sialosyl Lex, Ley, Lea, sialosyl Tn, Tn and fucosyl GM1, respectively. Paraffin-embedded tissue sections (30 squamous cell carcinomas, 30 adenocarcinomas and 27 small cell carcinomas) were tested by immunohistological staining. Evaluation of stained specimens was performed by taking mean value of scores evaluated by three independent examiners. In squamous cell carcinoma, expression of Ley, sialosyl Tn, sialosyl Lex and Lea was significantly higher than other antigens. Lex was also expressed especially in keratinized tissues. In adenocarcinoma, Lea was expressed most remarkably. Sialosyl Lex, Ley, sialosyl Tn were also highly expressed in malignant cells. There was no significant difference in staining patterns between well and poorly differentiated adenocarcinomas. Sialosyl Tn and sialosyl Lex were positive in morphologically normal mucous glands adjacent to tumors. In small cell carcinomas, Ley was expressed more than other types of tumors, whereas Lex and sialosyl Tn were less than others. Tn antigen was observed throughout adenocarcinomas, squamous and small cell carcinomas in a relatively weak manner. Dimeric Lex and fucosyl GM1 antigens were not detected. Most of normal lung sections showed negative staining with those MAbs. These findings indicate that there are differential expressions of certain carbohydrate antigens in different types of human lung cancers based on their origins.     

Oropharyngeal malignant epithelial cell, lymphocyte and macrophage CD44 surface receptors for hyaluronate are expressed in sustained EBV infection: Immunohistochemical data and EBV DNA tissue indices

The role of CD44 in Epstein-Barr virus (EBV)-related epithelial tumors is poorly understood. We studied the expression of CD44 in EBV infection in patients with oral squamous cell carcinoma (SCC) and nasopharyngeal carcinoma (NPC) and measured the EBV DNA. Whole blood, plasma and tissue samples from 8 male and 2 female patients with oral SCC, NPC, salivary gland lymphoepithelioma, normal salivary gland and buccal mucosa were assayed for EBV DNA. Expression of CD44, latent membrane protein (LMP), and labeling of lymphocytes, macrophages and dendritic cells were estimated by immunohistochemistry. Tissue EBV DNA was detected in 7 of 8 cases (87.5%) of oral malignant, benign and border-line lesions. LMP expression levels in tumors varied from absence and minimal to moderate - 50.3, 43.6, 6.0% and 91.1, 6.7, 2.2% for SCC and NPC, respectively. Levels of CD44 positivity in neoplasms were minimal (15.5 and 16.7%), moderate (30.3 and 47.8%), and diffuse (54.2 and 35.5%) for SCC and NPC, respectively, thus deviating from normal oral mucosa revealing heavily stained (100.0%) epithelial contours. CD19-positive B lymphocytes and S100-positive dendritic cells were intermixed with neoplastic cells. Collectively, CD44 mediated signaling may be implicated in EBV infection associated with the pathogenesis of oral SCC and NPC.     

Pediatric AIDS-Associated Lymphocytic Interstitial Pneumonia and Pulmonary Arterio-Occlusive Disease : Role of VCAM-1/VLA-4 Adhesion Pathway and Human Herpesviruses

Because the mechanisms of lymphocyte accumulation in the lungs of children with AIDS-associated lymphocytic interstitial pneumonia (LIP) are unknown, we studied the relative contributions of known adhesion pathways in mediating lymphocyte adherence to endothelium and the potential role of human herpesviruses in the expansion of these lesions. LIP was characterized by lymphoid hyperplasia of the bronchus-associated lymphoid tissue (BALT) and infiltration of the pulmonary interstitium with CD8(+) T lymphocytes. In some individuals there was expansion of the alveolar septae with dense aggregates of B lymphocytes, many containing the Epstein-Barr viral (EBV) genome. Patients with concurrent EBV infection also demonstrated large-vessel arteriopathy characterized by thickening of the intimae with collagen and smooth muscle. Venular endothelium from the lung of children with LIP, but not uninflamed lung from other children with AIDS or lung from children with nonspecific pneumonitis, expressed high levels of vascular cell adhesion molecule-1 (VCAM-1) protein. In turn, inflammatory cells expressing very late activation antigen-4 (VLA-4), the leukocyte ligand for VCAM-1, were the predominant perivascular infiltrate associated with vessels expressing VCAM-1. Expression of other endothelial adhesion molecules, including intracellular adhesion molecule-1 and E-selectin, was not uniformly associated with LIP. Using a tissue adhesion assay combined with immunohistochemistry for VCAM-1, we show that CD8(+) T cell clones that express VLA-4 bind preferentially to pulmonary vessels in sites of LIP: vessels that expressed high levels of VCAM-1. When tissues and cells were pretreated with antibodies to VCAM-1 or VLA-4, respectively, adhesion was inhibited by >/=80%. Thus, infiltration of alveolar septae with CD8(+) T cells was highly correlative with VCAM-1/VLA-4 adhesive interactions, and focal expansion of B cells was coincidental to co-infection with EBV.     

泛素特异性蛋白酶10在宫颈鳞状细胞癌中的表达及临床意义

目的:研究泛素特异性蛋白酶10(ubiquitin-specific protease 10,USP10)蛋白在人体宫颈鳞状细胞癌组织中的表达及临床意义.方法:采用免疫组织化学方法检测105例人体宫颈鳞状细胞癌及65例正常鳞状上皮组织石蜡切片中USP10蛋白的表达水平,并分析其表达与临床病理因素之间的相关性.结果:宫颈鳞状细胞癌组织中USP10阳性表达率为41.9%,而宫颈正常鳞状上皮组织中USP10的阳性表达率为61.5%.与正常鳞状上皮组织相比,USP10蛋白在宫颈鳞状细胞癌组织中的表达明显下调,差异具有统计学意义.同时,USP10蛋白表达与宫颈鳞状细胞癌的FIGO临床分期及肿瘤浸润深度呈负相关,而与E-cadherin,P53蛋白表达呈正相关.结论:宫颈鳞状细胞癌组织中USP10蛋白表达的下调与宫颈鳞状细胞癌的恶性程度及预后相关.     

Enhanced CD34 expression of sinusaid-like vascular endothelial cells in hepatocellular carcinoma

The immunohistochemlcal expression of CD34 (human hematopoletic stem cell and endothelial cell marker) and laminin were studied In chronic liver diseases and hepate cellular carcinoma (HCC) to elucidate whether their expression reflected phenotypic differences between non-cancerous slnusoids and sinusoid-like tumor vessels. In normal liver, hepatic sinusoids were always negative for CD34 and lami-nin. In chronic hepatitis and cirrhosis, the two antigens were sparsely expressed in capillarized sinusoids at periportal and petinodular area. In advanced HCC, CD34 was strongly and diffusely expressed by the endothelial lining of sinusoid-like tumor vessels. However, early-stage HCC showed a wide spectrum of CD34 expression from negative to focal and diffuse, strongly positive staining in sinusoid-like vessels. Laminin was strongly expressed in advanced HCC but not in early-stage HCC. The results Indicate that the enhanced expression of CD34 by sinusoidal endothelial cells may reflect the phenotypic change of endothellal cells in chronic liver diseases and HCC, and that the expression may correlate with the processes of angiogenesis induced by hepatocarcinogenesis.     

Expression and function of retinoblastoma binding protein 6 (RBBP6) in human lung cancer

Retinoblastoma binding protein 6 (RBBP6) interacts with both p53 and pRb, and has been identified as an E3 ubiquitin ligase due to the presence of a RING finger domain. RBBP6 promotes the degradation of p53, thereby increasing cell proliferation. However it is not known whether RBBP6 is expressed in lung cancer, or interacts with p53 and pRB to modulate the proliferation or apoptosis of lung cancer cells. As assessed by immunohistochemistry, RBBP6 and p53 proteins were overexpressed in lung adenocarcinomas and lung squamous cell carcinomas. Expression of RBBP6 mRNA in lung tumor tissue was demonstrated by quantitative RT-PCR and fluorescence in situ hybridization. Expression of RBBP6 mRNA was low in poorly differentiated tumors but high in well-differentiated tumors. Bronchoalveolar carcinomas showed intense RBBP6 mRNA hybridization in the cytoplasm of cells undergoing mitosis, supporting the association between RBBP6 expression and cell proliferation. By qRT-PCR, RBBP6 mRNA expression was 1.6 fold higher, whereas p53 mRNA expression was 2.9 fold lower in lung tumors compared with normal lung tissue. Transfection of lung adenocarcinoma and squamous cell carcinoma cells with RBBP6 siRNA decreased RBBP6 mRNA expression, whereas transfection with p53 siRNA increased RBBP6 mRNA expression. Treatment with RBBP6 siRNA increased the ratio of Bax/Bcl2 mRNA, suggesting that RBBP6 may have an anti-apoptotic function in lung cancer cells. This is the first report that RBBP6 mRNA and “its protein products” are expressed in subtypes of human lung cancer. RBBP6 may be involved in the degradation of p53, thereby enhancing cell proliferation and inhibiting apoptosis in lung cancer.