Desmoplastic reaction of gastric carcinoma: a light- and electron-microscopic immunohistochemical analysis using collagen type-specific antibodies

The desmoplastic reaction in ten cases of gastric carcinoma was investigated light and electron immunohistochemically by using monospecific antibodies to collagen types. In addition to type I and III collagens, type V collagen was constantly recognized in the fibrous stroma, increasingly of the scirrhous carcinoma. Type IV collagen delineated the basement membranes of carcinoma nests linearly with occasional discontinuity, whereas in the scirrhous carcinoma, it was present along the thick bundles of collagenous fibers. Immunoelectron microscopic studies revealed that type I and III collagens were distributed on the collagen fibers, and type V collagen was stained in the margin of these fibers. These antibodies also reacted in the rough endoplasmic reticulum of fibroblasts or myofibroblasts in a few cases. Type IV collagen was localized in the periphery of smooth muscle cells, endothelial cells of collapsed capillaries, and myofibroblasts scattered in the stroma of scirrhous carcinoma. Carcinoma cells were not reactive with any antibodies examined. These findings suggest that type V collagen, as well as type I and III collagens, is involved in the formation of desmoplastic stroma, and that these collagens are reactively synthesized by fibroblasts and myofibroblasts in some interaction with invading carcinoma cells.     

Cytophotometric analysis on nuclear DNA contents of human scirrhous gastric carcinoma

DNA ploidy patterns in six cases of scirrhous gastric carcinoma were examined by scanning cytophotometry, using 15 micron paraffin sections which were confirmed to involve a single whole nucleus. Measurements of DNA content were performed in 70 to 100 cancer cells each in intramucosal (m), submucosal (sm), propria muscular (pm) and subserosal (ss) parts of the tumor. Out of the six scirrhous gastric carcinomas, four were generalized ones and two were localized ones. Aneuploidy with diploid stem line (mosaic pattern) was seen in two cases (generalized type). The remaining four cases (two cases of localized type and two cases of generalized type) were composed of a diploid stem cell line with or without polyploid cells. Stem cell line did not change through the gastric wall, but in three cases (one diploid case and two mosaic cases), the number of polyploid or aneuploid cells significantly increased at the deep infiltrative site. These findings suggest that DNA ploidy of scirrhous gastric carcinoma is determined at an early stage of tumor proliferation and that the increase of polyploid cells and aneuploid cells in the deeper layer might participate in tumor progression. Moreover, there was no evident correlation between the increase of polyploid or aneuploid cells and the expression of oncogene products or EGF related growth factors.     

Detection of the epithelial membrane antigen in patients with varying status of the gastric mucosa using monoclonal antibodies

To study how the normal gastric mucosa develops chronic gastritis, gastric ulcer or cancer, biopsies from 103 patients with non-tumor gastric diseases and operative specimens from 12 patients with stomach carcinoma were examined for epithelial antigen (EA). In the peroxidase-antiperoxidase reaction performed on paraffin-embedded and cryostat sections of the gastric mucosa, monoclonal antibodies (MAb) to antigens located on the membranes of fatty breast milk globules were employed, which had been presented by the Department of Biomedical Sciences, Tampere University (Finland). The MAb 111C12-identified antigenic determinant was found to occur in 12.8% of the nonmalignant gastric mucosa examined and 48% of the stomach carcinoma one. The detection of EA is not characteristic of the given portion of the gastric mucosa in carcinoma.     

Integrin distribution in gastric carcinoma: Association of β3 and β5 integrins with tumor invasiveness

Immunolocalization of a variety of integrins using monoclonal antibodies against β3, β5, α3β1 and α6, and polyclonal antibodies against vitronectin receptor (αvβ3) and l were investigated on PLP-fixed paraffin sections of 19 cases of advanced gastric carcinomas. The β5 integrin, which pairs only with the av subunit, was positive in seven cases (37%), and was associated closely with scirrhous invasion (P < 0.05). β5 was positive chiefly in the cytoplasm of carcinoma cells and infrequently in cell membranes. β3, which is another subunit pairing with av, was positive in six cases (32%), and tended to be associated with scirrhous invasion (P < 0.1). β3 was also located chiefly in the cytoplasm. Five of the seven β5-positive cases showed coexpression of β3. Polyclonal antibodies to αvβ3 also showed a significant difference among the amounts of stroma (P < 0.05). Anti-β1 antibodies showed clear positivity in many cases (89%). Of the β1 integrins, α3β1 was positive in a few cases (26%) without any preferential pattern, and laminin receptor subunit α6 stained on cell membranes of neoplastic epithelia in many cases of carcinoma (89%) except for mucinous carcinoma. These distinctive patterns of integrin positivity indicate a close association of β5 and β3 expression with scirrhous invasion in gastric carcinoma.     

Abnormal distribution of collagen type IV in extrahepatic bile duct carcinoma

The present study investigated the pathogenesis of desmoplastic stroma formation, which is characteristic of most bile duct carcinomas and other scirrhous carcinomas. Using immunohistochemical analysis, the expression of collagen types I and IV, laminin and TGF-beta1 was examined in human extrahepatic bile duct carcinoma and compared with gastric and colon carcinoma. In addition to delineating the basement membranes of carcinoma nests and blood vessels, collagen type IV was present along the thick bundles of collagenous fibers in the stroma of extrahepatic bile duct carcinoma and scirrhous gastric carcinoma. The immunoreactivity of collagen type IV was strong in the adjacent or surrounding interstitium of tumor cell nests, but was absent or weak in older, more central portions of the tumor that contained sclerotic collagen. In situ hybridization demonstrated active expression of collagen alpha1(IV) mRNA in extrahepatic bile duct carcinoma and scirrhous gastric carcinoma cells. These results suggest that, although collagen type IV is typically a component of the basement membrane, it is expressed in the interstitial stroma of extrahepatic bile duct carcinoma and scirrhous gastric carcinoma where it may play a role in desmoplastic stroma formation.     

Distribution of collagen types I and III and basal lamina in human gastric carcinoma: An immunohistochemical and electron microscopic study

Summary Collagen types I and III were examined immunohistochemically in 32 cases of gastric carcinoma classified as poorly differentiated adenocarcinoma with scirrhous stroma, well differentiated adenocarcinoma with intermediate stroma, or poorly differentiated adenocarcinoma with medullary stroma. In the stroma of scirrhous carcinoma, types I and III collagens were distributed abundantly in fibrillar or granular patterns with little difference in the intensity of staining. In well differentiated adenocarcinoma, type I collagen was diffusely distributed in the stroma with type III collagen distributed sparsely. In poorly differentiated adenocarcinoma with medullary stroma, the two types of collagen were only found around capillaries, constituting the tumor interstitium. Electron microscopic examination of scirrhous carcinoma showed tumor cells partially covered with fibroblasts, and discontinuous basal lamina, collagen fibers and microfibrils present between tumor cells and fibroblasts. In well differentiated carcinoma, tumor cells were surrounded by fibroblasts, and well developed basal lamina was observed beneath the tumor cells. In poorly differentiated carcinoma with medullary stroma, the stroma consisted of capillaries and very few fibroblasts with discontinuous basal lamina occasionally being present between tumor cells and fibroblasts.     

Estrogen receptors in gastric adenocarcinoma: A retrospective immunohistochemical analysis

Summary Estrogen receptors (ER) in human gastric carcinomas were examined immunohistochemically using a specific monoclonal antibody to human ER. ER-immunoreactivity (ER-IR) was positive in 30 (27.8%) of the 108 gastric carcinomas examined. ER-IR was located in the nucleus of cancer cells. The incidence of ER-IR positive gastric carcinoma was not significantly different between male and female cases. However, the positive tumour cells were observed in 28 (39.4%) out of the 71 poorly differentiated adenocarcinoma, the incidence being significantly higher than that in well differentiated adenocarcinoma (p<0.01). There was no significant difference in the incidence of ER-IR between scirrhous carcinoma and non-scirrhous poorly differentiated adenocarcinoma. Synchronous expression of ER and epidermal growth factor receptor was found in 8 of the 26 scirrhous carcinomas (30.8%). Patients with ER-IR positive scirrhous gastric carcinomas showed a much worse prognosis than those with ER-IR negative scirrhous carcinomas.This work was supported in part by Grant-in Aid for Cancer Research No. 59-15 from the Ministry of Health and Welfare, Japan     

CUB-domain-containing protein 1 regulates peritoneal dissemination of gastric scirrhous carcinoma

CUB-domain-containing protein 1 (CDCP1) is a type-I transmembrane protein that is highly expressed in colon, breast, and lung cancers. We recently revealed that CDCP1 is associated with and phosphorylated by Src family kinases and is involved in the regulation of anchorage independence of certain lung cancer cell lines. In this study, we examined whether CDCP1 is involved in the regulation of tumor progression of scirrhous gastric cancer, which is a diffusely infiltrative carcinoma with high invasion potential. Expression and phosphorylation levels of CDCP1 correlated with the invasive potential of scirrhous gastric cancers. Reduction of CDCP1 expression by siRNA suppressed migration, invasion, and anchorage independence without affecting the proliferation of highly invasive scirrhous gastric cancer cells. However, CDCP1 overexpression promoted gastric cancer cell migration with low potential of invasion. Loss of CDCP1 suppressed invasion and dissemination of cancer cells that were orthotopically implanted in the gastric wall of nude mice. Expression and phosphorylation of CDCP1 were also detected in cancer cells of surgically resected tissues of human scirrhous gastric cancer by immunohistochemical analysis. Our results suggest that CDCP1 promotes invasion and peritoneal dissemination of cancer cells through the regulation of cell migration and anchorage independence. Therefore, it is both a potential prognostic and therapeutic target in certain types of gastrointestinal cancers, and suppression of its phosphorylation might be a useful strategy for modulating cancer metastasis.     

High expression of L-type amino-acid transporter 1 (LAT1) in gastric carcinomas: comparison with non-cancerous lesions

Amino acid transporters are essential for maintenance and proliferation of both normal and transformed cells. In the present study, L-type amino-acid transporter 1 (LAT1) immunoreactive expression was investigated in gastric carcinomas, in comparison with gastric adenomas and non-neoplastic lesions, using our recently developed novel monoclonal antibody. In a total of 87 cases of advanced gastric cancer, high LAT1 expression was observed in carcinoma cells, predominantly at plasma membranes with greater intensity in non-scirrhous than scirrhous carcinomas. Gastric carcinoma cases with lymph node metastasis showed significantly higher LAT1 expression than cases without lymph node metastasis. A positive correlation with Ki-67 LI was observed and the highly expressing non-scirrhous carcinomas showed a significantly poorer prognosis than the low LAT1 group. Cox hazard test revealed that TNM stage and LAT1 expression were independent prognostic factors in non-scirrhous carcinoma group. Further, a significant poor prognosis was confirmed in high LAT1 expression group, when limited to undifferentiated carcinoma cases excluding scirrhous carcinoma. Lower levels were found in adenomas. In conclusion, LAT 1 expression may be linked with cell proliferation and prognosis of gastric carcinomas, and offers a potential target for future anticancer therapy by inhibitors.     

The value of monoclonal antibody B72.3 for the diagnosis of breast carcinoma: experience with the first commercially available source.

One hundred cases of invasive breast carcinoma were studied using the commercially available monoclonal antibody Anti-Human Tumor-Associated Glycoprotein-72 (MAb B72.3, Biomedical Technologies Inc, Stoughton, MA) prediluted at 8.5 micrograms/mL. Forty-three cases displayed positive reactivity with this antibody. Intensity and distribution of positive staining varied among the tumor cells. Twenty-two cases had 1% or less reactive cells, while eight cases contained 40% or more positive tumor cells. Apical cell membrane and diffuse cytoplasmic staining were present. In fifteen cases intracytoplasmic lumina and extra-cellular secretory material were highlighted by positive staining. Thirty-five cases had benign breast tissue adjacent to the tumor. Benign ductal and lobular epithelial cells were nonreactive except for two cases in which small foci of apocrine metaplasia were positive. Reactivity with MAb B72.3 was not dependent upon histologic grade, nuclear grade, nodal status, or patient age. Excluding the lower number of positively stained cases, our findings were similar to other MAb B72.3 investigations. The number of positively stained cases and the intensity of the positivity were increased by using MAb B72.3 at 5.0 micrograms/mL with overnight incubation, or by using MAb B72.3 at 40.0 micrograms/mL with 2 hours of incubation. Our findings confirm that MAb B72.3 shows reliable reactivity with breast carcinoma that is sensitive to antibody concentration and incubation time without loss of specificity for tumor cells. Our results are also consistent with the view that MAb B72.3 probably detects epithelial membrane-related antigens in breast carcinoma, as do several other antibodies.     

Coexpression patterns of vimentin and glial filament protein with cytokeratins in the normal, hyperplastic, and neoplastic breast.

The authors studied by immunohistochemistry the intermediate filament (IF) protein profile of 66 frozen samples of breast tissue, including normal parenchyma, all variants of fibrocystic disease (FCD), fibroadenomas, cystosarcoma phylloides, and ductal and lobular carcinomas. Monoclonal antibodies (MAbs) to cytokeratins included MAb KA 1, which binds to polypeptide 5 in a complex with polypeptide 14 and recognizes preferentially myoepithelial cells; MAb KA4, which binds to polypeptides 14, 15, 16 and 19; individual MAbs to polypeptides 7, 13, and 16, 17, 18, and 19, and the MAb mixture AE1/AE3. The authors also applied three MAbs to vimentin (Vim), and three MAbs to glial filament protein (GFP). Selected samples were studied by double-label immunofluorescence microscopy and by staining sequential sections with some of the said MAbs, an MAb to alpha-smooth muscle actin, and well-characterized polyclonal antibodies for the possible coexpression of diverse types of cytoskeletal proteins. Gel electrophoresis and immunoblot analysis also were performed. All samples reacted for cytokeratins with MAbs AE1/AE3, although the reaction did not involve all cells. Monoclonal antibody KA4 stained preferentially the luminal-secretory cells in the normal breast and in FCD, whereas it stained the vast majority of cells in all carcinomas. Monoclonal antibody KA1 stained preferentially the basal-myoepithelial cells of the normal breast and FCD while staining tumor cell subpopulations in 4 of 31 carcinomas. Vimentin-positive cells were found in 8 of 12 normal breasts and in 12 of 20 FCD; in most cases, Vim-reactive cells appeared to be myoepithelial, but occasional luminal cells were also stained. Variable subpopulations of Vim-positive cells were noted in 9 of 20 ductal and in 1 of 7 lobular carcinomas. Glial filament protein-reactive cells were found in normal breast lobules and ducts and in 15 of 20 cases of FCD; with rare exceptions, GFP-reactivity was restricted to basally located, myoepithelial-appearing cells. Occasional GFP-reactive cells were found in 3 of 31 carcinomas. Evaluation of sequential sections and double-label immunofluorescence microscopy showed the coexpression of certain cytokeratins (possibly including polypeptides 14 and 17) with vimentin and alpha-smooth muscle actin together with GFP in some myoepithelial cells. The presence of GFP in myoepithelial cells was confirmed by gel electrophoresis and immunoblotting. Our results indicate that coexpression of cytokeratin with vimentin and/or GFP is comparatively frequent in normal basal-myoepithelial cells of the breast.(ABSTRACT TRUNCATED AT 400 WORDS)     

Kba.62 and S100 protein expression in cytologic samples of metastatic malignant melanoma

The diagnosis of melanoma can be challenging, especially in metastatic lesions, due to the ability of melanoma cells to morphologically mimic carcinoma, sarcoma and even lymphoma cells. Moreover, melanomas can exhibit negative immunostaining for the melanoma markers HMB‐45 and MART‐1/Melan‐A, often used in the diagnosis of this tumor. KBA.62 is a recently described antibody that reacts with benign and malignant melanocytic proliferations. In this study, we report our experience with KBA.62 and S100 protein immunostaining in the diagnosis of metastatic melanoma on fine‐needle aspiration and effusion samples. We reviewed 60 cytology samples from 58 patients with metastatic melanoma. Our results showed that KBA.62 stained 75% of the cases and S100 protein 87% of the cases. KBA.62 and S100 protein stained the majority of metastatic melanomas that were negative for HMB‐45 and MART‐1; KBA.62 stained 73% of the cases and S100 protein 73% of the cases. The majority (85%) of the cases negative for HMB‐45 and MART‐1 were positive for KBA.62 and/or S100 protein. Additionally, we also observed that KBA.62 staining was positive in the majority of epithelioid and spindle cell type melanoma cells. In conclusion, the performances of KBA.62 and S100 protein were similar and both markers are useful in the diagnosis of metastatic melanoma in cytology material, especially when the tumor cells lack expression of HMB‐45 and MART‐1. Diagn. Cytopathol. 2013;41:847‐851. © 2013 Wiley Periodicals, Inc.     

Immunohistochemical studies on the bovine lactating mammary gland (Bos taurus)

The study aimed to evaluate the validity of immunohistochemistry in the differential labeling of the diverse components of the lactating mammary gland. Paraffin-embedded sections of lactating bovine mammary glands were stained by conventional and histochemical techniques. Primary antibodies against S100, alpha smooth muscle actin (α-SMA), connexin-43 (Cx43), cytokeratin-14 (Ck14), galactosyltransferase (GalTase), angiotensin converting enzyme (ACE) and vascular endothelial growth factor (VEGF) were applied on paraffin sections. Strong cytoplasmic and nuclear S100 immunoreactivity was mainly expressed by alveolar epithelium and to a lesser variable extent by ductal epithelium. The Golgi zone of the epithelial cells expressed strong GalTase immunostaining. Myoepithelial cells displayed a strong immunostaining for α-SMA, Cx43 and Ck14, but not for S100. Vascular endothelium showed a moderate (for VEGF) to strong (for ACE) immunostaining. The presence of VEGF-immunoreactive mast cells within the interstitium may reflect their functional significance in angiogenesis, vascular permeability and migration of mononuclear leukocytes, suggesting their regulatory role in the secretory and immunological functions of the mammary glands.     

AMACR/34βE12/p63鸡尾酒双染对诊断前列腺癌及癌前病变的价值

目的探讨AMACR／34βE12／p63鸡尾酒双染对诊断小灶性前列腺癌及癌前病变的价值。方法从2005年6月起对3个月内临床连续送检的105例前列腺穿刺活检标本,6例前列腺癌根治标本和19例经尿道和耻骨上摘除的良性前列腺增生标本总计130个病例,1030个组织块中需要用免疫组织化学辅助诊断的262个组织块分别作AMACR／34βE12／p63鸡尾酒双染,同时作这3个抗体的单项染色,并结合HE切片和临床资料观察结果作出诊断。结果鸡尾酒双染切片中前列腺癌和高级别上皮内瘤变（HGPIN）上皮细胞呈蓝黑色,良性腺体的基底细胞呈红色。癌的蓝黑色腺上皮周围无红色基底细胞围绕,HGPIN的蓝黑色腺上皮周围有间断或连续的红色基底细胞。共在214个（82％）组织块中发现前列腺癌,包括31处小灶性癌。64个（24％）组织块中发现HGPIN,包括局灶性HGPIN和小腺泡性HGPIN。1个组织块有不典型性腺瘤样增生。未发现上皮细胞AMACR阳性同时基底细胞34βE12和p63阴性的良性腺体。结论鸡尾酒双染有助于提高小灶性前列腺癌和HGPIN检出率。     

Immunohistochemical detection of cd1a antigen in formalin-fixed and paraffin-embedded tissue sections with monoclonal antibody 010

The immunoreactivity of a CD1a monoclonal antibody (MAb), denoted 010, was investigated by means of the streptavidin-biotin-peroxidase method in formalin-fixed and paraffin-embedded tissues from 47 cases. The samples comprised reactive lymphoid proliferations of skin, tonsil, and lymph node including dermatopathic lymphadenopathy and Langerhans' cell histiocytosis, Hodgkin's and non-Hodgkin's lymphomas, and thymomas. Interdigitating and dermal dendritic cells, veiled cells, Langerhans' cells, and also cortical thymocytes and their neoplastic counterparts displayed immunostaining with MAb 010 in paraffin sections. These results are identical to previous ones reported for other CD1a MAbs in fresh or frozen specimens. The findings suggest that the binding site of 010 is a fixation-resistant epitope of CD1a antigen which has not been previously identified.     

Immunohistochemistry as an aid in the diagnosis of Hirschsprung's disease

The interpretation of rectal suction mucosal biopsies taken for the purpose of ruling out Hirschsprung's disease (HD) can be especially difficult in neonatal patients because of ganglion cell (GC) immaturity. Acetylcholinesterase histochemistry on frozen sections can be helpful but requires experience and may be complicated by excessive mucosal hemorrhage. The authors retrospectively have studied 27 patients, including 11 patients with HD, on whom conventionally fixed and embedded tissue was available, using an immunoperoxidase system directed against neuron-specific enolase (NSE) and S100 protein. NSE immunostaining produced intense staining of GC perikarya, greatly facilitating recognition of small immature forms. S100 immunostaining also highlighted GC as prominent negative stained cells surrounded by the positivity of Schwann cells. Both stains were helpful in demonstrating the overall pattern of microinnervation and its relationship to possible GC. The authors conclude that NSE and S100 immunostaining may facilitate interpretation of rectal mucosal biopsies when Hirschsprung's disease is being considered as a possibility.     

胃癌MT1-MMP、RECK蛋白的表达及其意义

目的探讨胃癌组织中膜型基质金属蛋白酶一1（MT1一MMP）和RECK蛋白表达状况和两者之间的关系,及其表达与胃癌临床诊断之间的关系。方法采用免疫组织化学法（两步法）对胃癌切除标本进行研究。结果在44例胃癌标本中,有37（84．1％）例MT1-MMP表达呈阳性反应,31（70．5％）例RECK免疫组化为阳性。MT1-MMP表达与胃癌分化具有一定的关系,低分化组织标本中,MT1一MMP表达较多,而中、高分化组织标本中,MT1-MMP表达相对较弱,具有统计学意义（P=0．015）。胃癌中MT1-MMP表达与肿瘤细胞浸润深度相关（P=0．007）。但MT1-MMP表达与性别以及有无淋巴结转移之间未见统计学差异。与此相反,在胃癌标本中,高分化标本中RECK蛋白表达相对较多,中分化标本其次,在低分化标本中表达较少。RECK表达与肿瘤分化具有统计学意义（P：0．006）。RECK表达与性别、肿瘤细胞浸润深度以及有无淋巴结转移无关,且与MT1-MMP表达之间亦无统计学意义。结论MT1-MMP过表达在胃癌分化、转移中发挥着重要的作用,而RECK的表达有益于胃癌向高分化发展。     

Early gastric stump carcinoma with rhabdoid features. Case report

Among 474 cases of gastric carcinoma studied in gastrectomy specimens from 1990 to 1996, only one (0.21%) showed positivity for vimentin. It was located on a gastric stump and, endoscopically, the tumor was classified as early gastric carcinoma type IIb + IIc. Histologically, tumor cells were extensively round to polygonal and had eosinophilic, or clear cytoplasm; the nuclei were large with conspicuous nucleoli. In some areas, the cytoplasm showed vimentin and(or cytokeratin coexpression by double immunostaining. Our results and the patient's rapid deterioration (death occurred six months after surgery) suggest that this type of tumor, although diagnosed as early carcinoma, behaved like an advanced malignancy.