正常胃肠道上皮粘液性质及其显示方法

应用组织化学方法包括PAS,AB/PAS,HID/AB,OR/AB,PAPS,mPAS,PB/KOH/PAS,PAT/KOH/PAS,PATB/KOH/PAS观察正常胃肠道粘膜分泌粘液的成分.对各种组织化学方法进行了比较,并对国内尚无报告的新方法进行了介绍.在正常胃粘膜表面上皮及胃窦腺体以分泌中性粘液为主.小肠主要分泌氮乙酰化唾液酸粘液,大肠上皮分泌以硫酸粘液及氧乙酰化唾液酸粘液为主.     

Mucinous (colloid) adenocarcinomas secrete distinct O-acylated forms of sialomucins: a histochemical study of gastric, colorectal and breast adenocarcinomas

AIMS: Mucinous (colloid) adenocarcinomas represent a distinct group of tumours defined by the presence of large amounts of extracellular mucins. By using histochemical methods, we analysed mucins secreted by mucinous versus non-mucinous adenocarcinomas and looked for differential secretion profiles. METHODS AND RESULTS: Sixty-four adenocarcinomas were studied (23 colorectal, 17 gastric, and 24 breast tumours). Thirty-two tumours were of the colloid type. The following methods were applied to paraffin tissue sections: (i) Alcian blue (pH 2.5) and periodic acid-Schiff (PAS); (ii) high iron diamine and Alcian blue (pH 2.5); (iii) periodic acid borohydride, potassium hydroxide, and PAS; (iv) periodic acid-thionine Schiff, potassium hydroxide, and PAS; and (v) periodic acid-borohydride and PAS. Most adenocarcinomas secreted acidic mucins, with sialomucins predominating over sulfomucins, except for non-mucinous adenocarcinomas of the breast which showed predominant neutral mucins. All mucinous adenocarcinomas contained C9-O-acyl sialic acid as mono, di(C8,C9)-, or tri(C7,C8,C9)-O-acyl forms. Acidic mucins secreted by the majority of non-colloid adenocarcinomas consisted of non-O-acylated sialomucins. CONCLUSIONS: C9-O-acylation of sialic acid is a characteristic feature of mucinous adenocarcinomas and can be readily detected by histochemical methods.     

O-acylated sialic acid variants in mucinous tumours of the ovary

Summary O-acylated sialic acid variants (site 8) can be demonstrated histochemically by the PB/KOH/PAS method. They are secreted by goblet cells of the lower gastrointestinal tract, by colorectal adenocarcinomas, and by their metastases. Since the metastases are positive only when the primary tumour is positive, O-acylated sialomucins can be considered to be specific markers of colorectal adenocarcinomas if identified in metastases of a tumour of unknown origin. In our histochemical study we evaluated 29 mucinous cystomas of the ovary (23 benign and 6 malignant). We found that six cases were positive to PB/KOH/PAS. The positivity was observed in a limited number of cells and only in areas which presented an intestinal type epithelium. It was also more evident in malignant cystomas than in benign ones. We therefore think that the PB/KOH/PAS positivity can not only be considered a marker of colorectal adenocarcinomas, but also of all neoplasms which originate from an intestinal epithelium or appear to an intestinal type epithelium.This work is partially supported by the M.P.I. (40%)     

A new histochemical technique of use in the interpretation and diagnosis of adenocarcinoma and villous lesions in the large intestine.

The periodic acid-thionin Schiff/potassium hydroxide/periodic acid-Schiff (PAT/KOH/PAS) procedure has been used to investigate the histochemical staining characteristics of the mucins found in adenocarcinoma and villous lesions of the large intestine. The 46 blocks examined represented 58 lesions from 37 patients, all of whom had had resections for carcinoma of the colon. tin sharp contrast to normal colon, none of the adenocarcinomas stained red with the PAT/KOH/PAS. With two exceptions the poorly and moderately differentiated adenocarcinomas stained blue, whereas of the well differentiated lesions half were blue and half purple. The malignant villous lesions demonstrated the same trends, although a larger percentage were purple. None of the benign lesions stained blue. It is suggested that malignancy in the colon is accompanied by an increase in blue staining in the PAT/KOH/PAS technique and that such staining may be of value in the interpretation of highly atypical adenoma where it might identify the onset of malignancy. This change in staining indicates a distinct alteration in the chemistry of the mucins which we interpret as a reduction in the degree of side chain O-acylation of their constituent sialic acids.     

FIBROBLAST GROWTH FACTOR 1 AND FIBROBLAST GROWTH FACTOR 2 IMMUNOREACTIVITY IN GASTROINTESTINAL TUMOURS

Acidic and basic fibroblast growth factors (FGF-1 and FGF-2) are mitogenic polypeptides that may play a role in autocrine and paracrine growth control of malignant tumours. We have examined the expression of FGF-1 and FGF-2 in a series of 41 colorectal tumours (24 adenomas, 17 adenocarcinomas) and 50 gastric adenocarcinomas (23 intestinal, 27 diffuse), using immunohistochemistry. Whereas the FGF-1 distribution was cytoplasmic, FGF-2 was restricted to the nuclei of the epithelial cells. FGF-1 immunoreactivity was detected in all samples (100 per cent), whereas FGF-2 immunoreactivity was seen in 17 adenomas (71 per cent), 13 colorectal carcinomas (76 per cent), and 29 gastric carcinomas (58 per cent). Compared with the normal mucosa, FGF-1 was overexpressed in 42 per cent of colorectal adenomas, 76 per cent of colorectal cancers, and 54 per cent of gastric cancers. Conversely, FGF-2 expression was reduced in 16 (66 per cent), 8 (47 per cent), and 40 (80 per cent) adenomas and colorectal and gastric samples, respectively. We found a significant correlation only between reduced FGF-2 and gastric tumour grade. These data indicate that FGF-1 overexpression occurs in a large proportion of human colorectal and gastric cancers. This may play a role in the progression of these tumours. The topographic variation in FGF-2 expression between normal (nuclear) and tumour (cytoplasmic) cells implies a corresponding functional change that may in turn facilitate tumour growth. © 1997 by John Wiley & Sons, Ltd.     

Expression of the 17-1A antigen in gastric and gastro-oesophageal junction adenocarcinomas: a potential immunotherapeutic target?

BACKGROUND: A murine monoclonal antibody against the 17-1A epithelial antigen has been shown to be a useful adjuvant therapy in colorectal cancer. Its clinical use could be extended to patients with upper gastrointestinal adenocarcinoma. AIM: To determine the distribution of the antigen in gastric and oesophageal adenocarcinoma. METHODS: The activity of two monoclonal antibodies active against 17-1A epithelial antigen was studied in gastric and gastro-oesophageal junction adenocarcinomas: fresh frozen tissue from both the carcinoma and adjacent mucosa was stained using immunocytochemistry with a murine monoclonal antibody (17-1A edrecolomab, Glaxo Wellcome); paraffin embedded tissue was stained using the humanised monoclonal antibody 3622W94 (Glaxo Wellcome). RESULTS: 29 of 33 cancers (88%) stained with the murine antibody and 39 of 40 (98%) with the humanised antibody. The degree of staining was greater in well differentiated and moderately differentiated tumours. There was no staining of the normal background gastric or oesophageal mucosa, but areas of intestinal metaplasia stained intensely. The humanised monoclonal 3622W94 antibody produced more intense staining than the murine antibody. CONCLUSIONS: The high incidence of expression of the 17-1A antigen in patients with gastric and gastro-oesophageal junction adenocarcinomas suggests a potential role for these antibodies as an adjuvant treatment for these common cancers.     

A study of vesical adenocarcinoma, intestinal metaplasia and related lesions using mucin histochemistry

Biopsy and autopsy material from the urinary bladder was studied using PAS and PAS-D techniques to identify glycogen and neutral mucins, the alcian blue/high iron diamine method to distinguish sialo- and sulphamucins and the PB/KOH/PAS technique to localize O-acylated sialomucins. All of 10 examples of normal urothelium and both of two cases of transitional carcinoma in situ contained glycogen, but no mucin. Other lesions displayed one of two patterns of mucin production: the extracellular mucin seen focally in 17 cases of cystitis cystica consisted of sialo- and/or neutral mucins only, a pattern also displayed by mucins produced in 10 of 13 examples of transitional cell carcinomas and by three of nine tumours purely or in part adenocarcinomas. The intracellular mucins expressed in five of the 17 cases of cystitis glandularis and in all of eight cases of frank intestinal metaplasia with goblet cells displayed a colonic phenotype, with production of O-acylated sialomucins. A similar profile was expressed by six adenocarcinomas and this included tumours likely to be of vesical and also of urachal origin. It is concluded that identification of O-acylated mucins cannot distinguish between primary bladder tumours and metastases from a colonic primary, or between carcinomas of vesical and urachal origin.     

Failure to demonstrate specificity of the morphological and histochemical changes in mucosa adjacent to colonic carcinoma (transitional mucosa).

Characteristic morphological and histochemical changes in mucosa adjacent to colorectal adenocarcinomas have been described. It has been suggested that this type of mucosa, labelled transitional mucosa (TM) because of its specific association with colorectal adenocarcinoma, is indicative of a premalignant change. In an investigation of mucosa adjacent to anal tumours extending into the rectum and mucosa from solitary ulcer syndrome and colostomies the mucosal alterations described in TM could be demonstrated. Thus TM is not specifically related to colorectal adenocarcinoma and probably arises as a secondary phenomenon. The claim that TM represents a premalignant change cannot be substantiated.     

AMACR is highly expressed in gastric adenomas and intestinal-type carcinomas

Alpha-methylacyl-CoA racemase (AMACR) is a novel tumor biomarker expressed in a number of neoplasms, including colorectal and prostatic adenocarcinomas. However, AMACR expression has not been investigated in preneoplastic and neoplastic lesions of the stomach. Using immunohistochemistry we studied the expression of AMACR in normal gastric mucosa (n=32), intestinal metaplasia (n=26), adenomas (n=29) and adenocarcinomas (n=132) of the stomach from 135 patients. Synchronous adenocarcinomas arising in the background of adenomas were observed in 26 cases. AMACR immunoreactivity was not observed in all normal gastric mucosa. Tissue from intestinal metaplasia, adenomas, and adenocarcinomas was positive in 7.7% (2/26), 79.3% (23/29), and 62.9% (83/132) of cases, respectively. The difference in AMACR expression between adenomas or adenocarcinomas and non-neoplastic mucosa was statistically significant (p=0.0001). Moreover, intestinal-type carcinomas showed significantly higher expression of AMACR (69.8%) compared to diffuse-type carcinomas (47.2%) (p=0.02). Our results indicate that as well as being an additional diagnostic tool, altered AMACR expression in gastric adenomas and intestinal-type carcinomas suggests that AMACR may be involved early in the development of intestinal-type gastric carcinomas.     

CDX-2 and HER-3 expression in canine gastric and colorectal adenocarcinomas

CDX-2 is used as a specific cell marker for human intestinal adenocarcinoma. In human studies, HER-3 overexpression predicts poor survival for patients with various cancers including gastric cancer. Gastrointestinal adenocarcinoma is less common in dogs than in man and the expression of immunological markers by the canine tumours has not yet been extensively studied. CDX-2 and HER-3 expression was determined in 18 canine gastrointestinal adenocarcinomas: 13 were of colorectal origin and five were of gastric origin. CDX-2 expression was predominantly observed in the nuclei of normal colonic epithelium and in neoplastic epithelium and neoplastic gastric epithelial cells that which had metastasized to the gastric lymph node. CDX-2 was expressed in 11 of 13 (84.6%) colorectal adenocarcinomas and in all five (100%) gastric adenocarcinomas. HER-3 was consistently expressed in the cytoplasm of neoplastic epithelial cells. HER-3 expression was detected in 12 of 13 (92.3%) colorectal and in all five (100%) gastric adenocarcinomas. CDX-2 and HER-3 may be useful markers for canine gastrointestinal adenocarcinoma.     

Mucin profiles in the mucosa adjacent to large bowel non-adenocarcinoma neoplasias

Morphological features and mucin secretion patterns were investigated in the colonic mucosa adjacent to or overlying mesenchymal or primary epithelial neoplasias, other than adenomas and adenocarcinomas. The material included 15 cases of non-adenocarcinoma tumours examined during 1978-1981. Increased sialomucins and morphological features similar to those described in the so-called 'transitional' mucosa adjacent to primary colorectal adenocarcinomas were observed in only two cases. In contrast our previous studies have demonstrated 'transitional' profiles in 98% of adenocarcinomas.     

Differentiation towards gastric foveolar, mucopeptic and intestinal goblet cells in gallbladder adenocarcinomas

Cellular differentiation in 22 surgically removed adenocarcinomas of the gallbladder was immunohistochemically studied with antibodies specific to mucins of gastric foveolar cells (M1), (pseudo)pyloric cells (M2) and intestinal goblet cells (M3), and also with antibodies against pepsinogen II and chromogranin A. More than 70% of tumours (16 of 22 cases) displayed gastric-and/or intestinal-type differentiation, most of which (12 of 16 cases) showed both types of differentiation. Two tumours showed an organoid growth pattern similar to the normal gastric mucosa. The presence of endocrine cells positive for chromogranin A was closely related to that of gastric- and/or intestinal-type cells. The present findings clearly indicate the multidirectional differentiation of gallbladder adenocarcinomas and suggest that most gallbladder adenocarcinomas develop and progress under induction of gastric and intestinal differentiation.     

Phenotypic change of muscularis mucosae in early invasive colorectal adenocarcinoma

BACKGROUND: Invasive colorectal adenocarcinomas have bundles of eosinophilic spindle cells, which are regarded as myofibroblasts, in their desmoplastic stroma, some of which are continuous with the muscularis mucosa. AIM: To investigate the relation between the eosinophilic spindle cells and the muscularis mucosa based on their cytoskeletal phenotypes in early invasive colorectal adenocarcinoma. METHODS: Formalin fixed, paraffin wax embedded tissues of 17 early invasive colorectal adenocarcinomas were immunostained for alpha-smooth muscle actin (alpha-SMA), desmin, and vimentin. RESULTS: The phenotype of the muscularis mucosa was alpha-SMA positive, desmin positive, and vimentin weakly positive, whereas the eosinophilic spindle cells showed a decreased degree of immunoreactivity for alpha-SMA and desmin in particular, and an increased degree of immunoreactivity for vimentin. The degree of phenotypic difference between the muscularis mucosa and the eosinophilic spindle cells was greater in the eosinophilic spindle cells in the centre of the invasive area that were not continuous with the muscularis mucosa than in the eosinophilic spindle cells continuous with the muscularis mucosa. CONCLUSIONS: These findings suggest that the smooth muscle cells of the muscularis mucosa change their phenotype to become eosinophilic spindle cells, namely myofibroblasts, in the early invasive area of colorectal adenocarcinoma.     

Expression of c-erbB-2 oncogene product in Barrett's adenocarcinoma: pathological and prognostic correlations.

AIMS--To establish the prevalence of c-erbB-2 protein expression in a surgical series of Barrett''s adenocarcinomas; and to correlate this expression with clinicopathological data and prognosis. METHODS--Sixty six surgical specimens of Barrett''s adenocarcinomas were included in this retrospective study. Blocks of the tumour and of non-dysplastic Barrett''s mucosa were stained with a polyclonal antibody specific for the intracytoplasmic domain of the c-erbB-2 protein. RESULTS--Seven of 66 tumours showed membrane staining for the c-erbB-2 protein. The non-dysplastic Barrett''s mucosa was negative in all cases. There was no difference between c-erbB-2 positive and negative tumours with regard to mean age, sex ratio, percentage of alcohol misusers, percentage of smokers, tumour differentiation, depth of invasion, lymph node response, and proliferative activity, assessed by the percentage of tumour cells positive with the MIB-1 antibody directed against the Ki-67 antigen. All c-erb B2 positive tumours were of Lauren''s intestinal type compared with negative c-erbB-2 tumours. Patients with c-erbB-2 positive tumours had a significantly poorer prognosis than patients with negative tumours. CONCLUSIONS--The prevalence of Barrett''s adenocarcinomas expressing c-erbB-2 found in this study (11%) was similar to that observed in published series of gastric adenocarcinomas. c-erbB-2 protein expression could be an important prognostic indicator in Barrett''s adenocarcinoma.     

Epithelial markers in prostatic, bladder, and colorectal cancer: an immunoperoxidase study of epithelial membrane antigen, carcinoembryonic antigen, and prostatic acid phosphatase

Twenty prostatic adenocarcinomas, 20 transitional cell carcinomas of the bladder, and 20 colorectal adenocarcinomas were stained for epithelial membrane antigen, carcinoembryonic antigen, and prostatic acid phosphatase. Polyclonal affinity purified first and second antibodies and an indirect immunoperoxidase technique were used. All of the colorectal and bladder tumours and 16/20 prostatic tumours were positive for epithelial membrane antigen. All 20 colorectal, 7/20 bladder, and 5/20 prostatic tumours stained for carcinoembryonic antigen. All of the prostatic adenocarcinomas and none of the colorectal or bladder tumours were positive for prostatic acid phosphatase. These markers may be used to discriminate between tumours arising from these sites.     

347例大肠息肉及其癌变的粘液组化和免疫组化观察

对大肠息肉及其癌变347例的粘液组化和223例的免疫组化研究显示,它们的粘液分泌改变和四种结肠癌相关抗原表达与其组织学类型、体积大小和不典型增生程度相关,并且PAT/KOH/PAS粘液组化染色显示氮乙酰化、侧链C_7、C_9位氧乙酰化唾液酸粘液分泌可能代表了息肉的恶变倾向,结肠癌单克隆抗体CL-2、CL-4的免疫组化染色对判断腺瘤癌变有较大参考价值。     

Differential expression of vesicular monoamine transporter (VMAT) 1 and 2 in gastrointestinal endocrine tumours.

Neuroendocrine tumours are characterized by their capacity to produce hormones, which are stored in vesicles and secretory granules. Demonstration of granule/vesicle proteins in tumours is taken as evidence of neuroendocrine differentiation. Vesicular monoamine transporters (VMAT1 and VMAT2) mediate the transport of amines into vesicles of neurons and endocrine cells. The expression of VMAT1 and VMAT2 and the usefulness of VMAT1 and VMAT2 in the histopathological diagnosis of gastrointestinal endocrine tumours have not been fully explored. This study therefore investigated the expression of VMAT1 and VMAT2 in 211 human gastrointestinal tumours by immunocytochemistry and western blotting. VMAT1 and/or VMAT2 were demonstrated in the majority of amine-producing endocrine tumours of gastric, ileal, and appendiceal origin. Serotonin-producing endocrine tumours (ileal and appendiceal carcinoids) expressed predominantly VMAT1, while histamine-producing endocrine tumours (gastric carcinoids) expressed VMAT2 almost exclusively. In peptide-producing endocrine tumours such as rectal carcinoids and endocrine pancreatic tumours, only a small number of immunopositive tumour cells were observed. No labelling was found in non-endocrine tumours, including gastric, colorectal and pancreatic adenocarcinomas and gastrointestinal stromal tumours. In conclusion, VMAT1 and VMAT2 are differentially expressed by gastrointestinal endocrine tumours, with a pattern specific for each tumour type, reflecting their neuroendocrine differentiation and origin. VMAT1 and VMAT2 may therefore become valuable markers in the classification of neuroendocrine tumours and may also indicate patients suitable for radioisotope treatment operating via these transporter systems.     

Mucin 16 (cancer antigen 125) expression in human tissues and cell lines and correlation with clinical outcome in adenocarcinomas of the pancreas, esophagus, stomach, and colon

Mucin 16 (cancer antigen 125) is a cell surface glycoprotein that plays a role in promoting cancer cell growth in ovarian cancer. The aims of this study were to examine mucin 16 expression in a large number of digestive tract adenocarcinomas and precursors and to determine whether mucin 16 up-regulation is correlated with patient outcome. Tissue microarrays were constructed using surgical resection tissues and included pancreatic (115 normal, 29 precursors, 200 pancreatic ductal adenocarcinomas), esophageal (86 normal, 104 precursors, 95 esophageal adenocarcinomas, 35 lymph node metastases), gastric (211 normal, 8 precursors, 119 gastric adenocarcinomas, 62 lymph node metastases), and colorectal (34 normal, 17 precursors, 39 colorectal adenocarcinomas) tissues. Mucin 16 was detected in 81.5%, 69.9%, 41.2%, and 64.1% of the pancreatic ductal adenocarcinomas, esophageal adenocarcinomas, gastric adenocarcinomas, and colorectal adenocarcinomas, respectively. Mucin 16 was seen in a subset of the precursors. On multivariate analysis, moderate/diffuse mucin 16 in pancreatic ductal adenocarcinomas was strongly associated with poor survival (P < .001), independent of other prognosis predictors. A similar trend was observed for esophageal adenocarcinomas (P = .160) and gastric adenocarcinomas (P = .080). Focal mucin 16 in colorectal adenocarcinomas was significantly correlated (P = .044) with a better patient outcome, when compared with mucin 16-negative cases. Using Western blot analysis, we found mucin 16 expression in 3 of 6 pancreatic ductal adenocarcinoma and 1 of 2 esophageal adenocarcinoma cell lines. We conclude that most of the digestive tract adenocarcinomas and a subset of their precursors express mucin 16. Mucin 16 expression is an independent predictor of poor outcome in pancreatic ductal adenocarcinomas and potentially in esophageal adenocarcinomas and gastric adenocarcinomas. We propose that mucin 16 may function as a prognostic marker and therapeutic target in the future.     

Guanylyl cyclase C is a specific marker for differentiating primary and metastatic ovarian mucinous neoplasms

Aims:  The aim was to assess the value of GCC in distinguishing primary ovarian mucinous neoplasms from metastatic mucinous adenocarcinomas with ovarian involvement. Guanylyl cyclase C (GCC) is a brush border membrane receptor for the endogenous peptides guanylin and uroguanylin, and the homologous diarrhoeagenic bacterial heat-stable enterotoxins that is selectively expressed by epithelial cells from the duodenum to the rectum, but not by normal epithelia of the stomach or oesophagus, or normal extramucosal cells in humans.
Methods and results:  Fifty ovarian tumours: 27 primary ovarian mucinous neoplasms (seven cystadenomas, 10 borderline tumours and 10 cystadenocarcinomas) and 23 metastatic mucinous adenocarcinomas with ovarian involvement [13 colorectal adenocarcinomas, four gastric adenocarcinomas, six appendiceal mucinous tumours (four adenocarcinomas, one with neuroendocrine features, and two appendiceal mucinous cystadenomas)] were studied. For primary ovarian mucinous neoplasms, 25 of 27 were negative for GCC. Twelve of 13 cases of colorectal adenocarcinoma (except for one neuroendocrine adenocarcinoma) were positive for GCC. Three of four appendiceal mucinous adenocarcinomas were positive for GCC in both the primary and metastatic tumours (except for one neuroendocrine adenocarcinoma). Two of two appendiceal mucinous cystadenomas were positive for GCC. Of four cases of gastric adenocarcinoma with ovarian involvement, only one (primary tumour) exhibited focal GCC staining.
Conclusions:  GCC is a useful marker for differentiating between primary and secondary ovarian mucinous neoplasms.