The relationship of quantitative nuclear morphology to molecular genetic alterations in the adenoma-carcinoma sequence of the large bowel.

The relationship of abnormal nuclear morphology to molecular genetic alterations that are important in colorectal tumorigenesis is unknown. Therefore, Feulgen-stained isolated nuclei from 22 adenomas and 42 carcinomas that had been analyzed for ras gene mutations and allelic deletions on chromosomes 5q, 18q, and 17p were characterized by computerized image analysis. Both nuclear area and the nuclear shape factor representing irregularity correlated with adenoma-carcinoma progression (r = 0.57 and r = 0.52, P < 0.0001), whereas standard nuclear texture, a parameter of chromatin homogeneity, was inversely correlated with progression (r = -0.80, P < 0.0001). The nuclear parameters were strongly interrelated (P < 0.0005). In multivariate analysis, the nuclear parameters were predominantly associated with adenoma-carcinoma progression (P < or = 0.0001) and were not influenced significantly by the individual molecular genetic alterations. Nuclear texture, however, was inversely correlated with fractional allelic loss, a global measure of genetic changes, in carcinomas (r = -0.39, P = 0.011). The findings indicate that nuclear morphology in colorectal neoplasms is strongly related to tumor progression. Nuclear morphology and biologic behavior appear to be influenced by accumulated alterations in cancer-associated genes.     

Morphometrical analysis of gall-bladder adenoma and adenocarcinoma with reference to histogenesis and adenoma-carcinoma sequence

Summary In order to examine whether our subdivision of gall-bladder adenoma and adenocarcinoma into nonmetaplastic and metaplastic types is reasonable from the viewpoint of their cytological features, a morphometrical analysis was conducted on 17 adenomas and 59 adenocarcinomas. The morphometrical parameters used were nucleo-cytoplasmic ratio (N/C ratio) and nuclear areas (NA). N/C ratio in the metaplastic type of both adenoma and adenocarcinoma was significantly larger than that in the non-metaplastic. This result shows that the different tumour types are associated with a different N/C ratio. Continuous measurement of N/C ratio and NA in progressing from non-cancerous mucosa to the lesion was made and the data obtained were analysed by the Lowess method. In some adenomas the total area of polypoid lesions was serially measured and these data were also analysed by the Lowess method. The results showed different processes in non-metaplastic and metaplastic types of adenoma and adenocarcinoma from the standpoint of nuclear changes of N/C ratio and NA. These results indicate that our histogenetic classification of adenocarcinoma is reasonable in morphometrical nuclear analysis. We also investigated the adenoma-carcinoma sequence as a possible histogenesis for gall-bladder carcinoma. Eight (72.7%) of 11 metaplastic adenomas and two (33.3%) of six non-metaplastic adenomas had foci of atypical gland proliferation and were considered to be carcinomas. Moreover, these carcinomatous areas were surrounded by severe dysplasia. These findings indicate that adenoma-carcinoma sequence accounts for one of the histogeneses of gall-bladder carcinoma.     

The biological and prognostic significance of cell polarity and E-cadherin in grade I infiltrating ductal carcinoma of the breast

Grading of breast cancer based on the modified Scarff, Bloom, and Richardson system provides invaluable prognostic information. Recent evidence suggests that most tumours do not usually progress between grades and that groups of tumours within each grade are biologically distinct. This study has explored one potential aspect of biological tumour heterogeneity within grade by examining the relationship between cell polarity, the cell adhesion molecule E-cadherin, a major effector of cell polarity, and outcome, in 149 grade I infiltrating ductal breast carcinomas. Polarity was evaluated by studying the degree to which three features of polarized epithelial cells—nuclear ordering, basal positioning of nuclei within cells, and apical snouting/blebbing—were present in these tumours. E-cadherin expression was investigated using the antibody HECD-1. A low degree of tubule formation was correlated with poor nuclear ordering ( p< 0·01). The three histological features—nuclear ordering, basal nuclei, and apical blebbing—were all correlated with each other (all p< 0·0001). Polarity measurements did not correlate with survival. E-cadherin expression did not correlate with polarity and negative tumours were still able to form tubules. Surprisingly, strong E-cadherin immunostaining correlated with poor survival, tumour size, and nodal status. On univariate parametric (Weibull) survival models, high E-cadherin scores and tumour size were both significant predictors of survival in this group. Copyright © 1999 John Wiley & Sons, Ltd.     

Matrix metalloproteinases and E-cadherin immunoreactivity in different basal cell carcinoma histological types

The immunohistochemical staining of matrix metalloproteinases (MMPs) and E-cadherin in tumor epithelial and stromal cells was analyzed in a group of solid, superficial spreading and cystic tumors and in a group of morpheaform and recurrent basal cell carcinomas (BCC) in order to determine whether any of these factors possibly contribute to tumor therapy resistance. Tumor tissues of 64 patients were obtained by complete excisional or curettage biopsy of BCC and these were immunohistochemically stained for MMP-1, MMP-2, MMP-9, MMP-13 and E-cadherin. In the morpheaform and recurrent BCC, MMP-9 expression significantly increased in the stroma, while E-cadherin expression was negative in epithelial cells. Odds ratio for development of morpheaform and recurrent BCC was 6.2 for positive MMP-1 immunostaining in epithelial tumor cells, 5.8 for positive MMP-9 immunostaining in tumor stroma, 3.2 for positive MMP-13 immunostaining in tumor stroma, and 4.5 for negative E-cadherin in epithelial tumor cells. Our results suggest that MMP-1 immunostaining in tumor cells, MMP-9 expression in stromal cells, and absence of E-cadherin expression are associated with morpheaform and recurrent BCC.     

Expression of catenins and E-cadherin during epithelial restitution in inflammatory bowel disease

Catenins are cytoplasmic proteins associated with E-cadherin, the prime mediator of cell–cell adhesion. Perturbation in any of these molecules results in altered intercellular adhesion, cell differentiation, and increased migration. In this study, the expression and cellular localization of catenins and E-cadherin in inflammatory bowel disease were examined. The expression of E-cadherin; α-, β-, and γ-catenin; and p120 was evaluated immunohistochemically in 31 paraffin-embedded colonic specimens from 21 patients with ulcerative colitis and Crohn's disease. Loss of normal membranous E-cadherin and α-catenin staining was detected at the mucosal edges around epithelial ulcerations in all cases of active ulcerative colitis and in 50 per cent of cases with active Crohn's disease. Reduced expression of p120 protein was also found at the margins of ulcerated mucosa in all cases of active ulcerative colitis and in 75 per cent of those with active Crohn's disease. There was a statistically significant correlation between the expression of E-cadherin, α-catenin and p120 and disease activity. There were no changes in β- and γ-catenin expression in either ulcerative colitis on Crohn's disease. These findings indicate that altered expression of E-cadherin, α-catenin, and p120 occurs during mucosal ulceration in inflammatory bowel disease. These changes may be involved in promoting cell migration during epithelial restitution of the gastrointestinal mucosa. © 1998 John Wiley & Sons, Ltd.     

E-钙粘蛋白在子宫腺肌病组织中的表达

目的　检查E 钙粘蛋白 (E cadherin ,E cad)在子宫腺肌病病灶中的分布及其mRNA的表达 ,以探讨其在子宫腺肌病发病机制中的作用。方法　采用免疫组化二步法和原位杂交技术 ,检查E -cad在子宫腺肌病病灶中的表达及其mRNA的转录水平 ,并以正常子宫内膜作对照。采用图像分析技术检测E -cad及其mRNA表达的平均光密度值。结果　免疫组化显示 ,在正常的内膜中E -cad主要表达在腺上皮细胞的细胞膜 ,以侧面膜为主 ;在腺肌病病灶的表达明显减少 (P <0 .0 1)。原位杂交显示 ,在腺肌病病灶和正常内膜中 ,E -cad的mRNA均表达于腺上皮细胞的胞浆 ,但在后者中的表达强度明显高于前者 ,其平均光密度值有显著性差异 (P <0 .0 1)。结论　E 钙粘蛋白在腺肌病病灶中低表达及其mRNA的低转录水平是子宫腺肌病发病机制中的一个较重要的因素。     

Loss of extracellular E-cadherin in the normal mucosa of duodenum and colon of patients with familial adenomatous polyposis

The duodenum is the main site for (pre-) malignant extracolonic manifestations in patients with familial adenomatous polyposis (FAP). Changes in the E-cadherin/beta-catenin complex play a pivotal role in the development of malignancies. Loss of E-cadherin has been described in association with loss of SMAD4. To elucidate the pathways leading to the development of duodenal adenomas in patients with FAP, the distributions of E-cadherin, SMAD4, and beta-catenin were analyzed. Furthermore, differences between the duodenum and colon were evaluated. Normal FAP duodenum (n = 13) and FAP duodenal adenomas (n = 50; total, 21 patients) were compared with non-FAP duodenal adenomas (n = 7) and normal non-FAP duodenum (n = 15) by immunohistochemical staining for extracellular and intracellular E-cadherin, beta-catenin, and SMAD4. Colonic biopsies of 10 patients with FAP were also studied, as well as non-FAP colonic adenomas (n = 26) and non-FAP normal colon (n = 12). Compared with the intracellular component of E-cadherin that was present in all cases, a significant loss of extracellular E-cadherin was observed in both duodenal and colonic adenomas and normal tissue of patients with FAP. Nuclear localization of beta-catenin was more often observed in duodenal FAP adenomas compared with non-FAP adenomas. Loss of nuclear SMAD4 was seen in the duodenum and, to a higher degree, in the colon of patients with FAP, as well as non-FAP patients. The loss of extracellular E-cadherin in the normal duodenal and colonic mucosa of patients with FAP might play a role in the high susceptibility of these tissues for (pre-) malignant transformation.     

Effect of indomethacin on E-cadherin and beta-catenin expression in HT-29 colon cancer cells

Nonsteroidal anti-inflammatory drugs (NSAIDs) lower the incidence of and mortality from colon cancer. Although there is much evidence from epidemiological and laboratory studies that NSAIDs have antitumor activity and reduce the incidence of colon cancer, the mechanism of action remains unknown. In this paper, we present the effect of indomethacin on growth inhibition, induction of apoptosis, and alterations in the expression of several genes involved in Wnt signaling in HT-29 colon cancer cells. We have shown that indomethacin reduces the proliferation rate of HT-29 colon cancer cells and induces apoptosis. Concentrations of indomethacin from 10(-4) to 10(-3) M strongly inhibited the growth of HT-29 cells. The inhibition of growth, as well as induction of apoptosis was dose and time dependent. The treatment of cells with 4 x 10(-4) M indomethacin caused strong inhibition of cell growth (about 70%), enhanced expression of APC, decreased expression of beta-catenin and induced expression of E-cadherin proteins. Expression of beta-catenin was not markedly reduced instead, beta-catenin was translocated from the nucleus and cytoplasm to the plasma membrane. These results were confirmed by real-time RT-PCR analysis on mRNA level. At a concentration of 4 x 10(-4) M indomethacin there was increased expression of APC gene (10.9-fold induction; DeltaDeltaCt = 3.43) and E-cadherin gene (3.5-fold induction; DeltaDeltaCt = 1.79). These results suggest the antiproliferative effect of indomethacin may contribute to enhanced cell adhesion through increased expression of E-cadherin and translocation of beta-catenin from the nucleus to the cell membrane.     

Microsatellite instability in the adenoma-carcinoma sequence of the stomach

Sixty-three cases of stomach resections harboring both adenoma and carcinoma were analyzed for microsatellite instability (MSI). The cases included 28 carcinomas arising from adenoma (Type I) and 35 carcinomas with separate adenoma (Type II). The results of MSI assessed by 49 markers were the same for BAT-26 instability. The incidence of MSI was 21% in gastric adenoma and 30% in gastric carcinoma, which is significantly higher than gastric carcinoma without associated adenoma (p < 0.01). Five of eight (63%) cases of multiple carcinomas associated with adenoma showed MSI+ in adenoma and in one or more carcinoma lesion(s). Eight of thirteen (62%) MSI+ adenomas were associated with carcinoma, whereas 20 of 50 (40%) MSI adenomas were associated with carcinoma. MSI+ adenomas of Type I showed a higher mutation rate of the TGF-beta RII gene than Type II (88% versus 40%). Gastric adenoma with TGF-beta RII gene mutation was more prone to transform into carcinoma (p = 0.03). This study revealed that gastric carcinoma arising from adenoma is frequently associated with a mismatch repair deficiency mechanism. In the gastric adenoma-carcinoma sequence, TGF-beta RII gene mutation occurred early in the adenoma stage and it persisted after malignant transformation.     

The importance of E-cadherin binding partners to evaluate the pathogenicity of E-cadherin missense mutations associated to HDGC

In hereditary diffuse gastric cancer (HDGC), CDH1 germline gene alterations are causative events in 30% of the cases. In 20% of HDGC families, CDH1 germline mutations are of the missense type and the mutation carriers constitute a problem in terms of genetic counseling and surveillance. To access the pathogenic relevance of missense mutations, we have previously developed an in vitro method to functionally characterize them. Pathogenic E-cadherin missense mutants fail to aggregate and become more invasive, in comparison with cells expressing the wild-type (WT) protein. Herein, our aim was to develop a complementary method to unravel the pathogenic significance of E-cadherin missense mutations. We used cells stably expressing WT E-cadherin and seven HDGC-associated mutations (five intracellular and two extracellular) and studied by proximity ligation assays (PLA) how these mutants bind to fundamental regulators of E-cadherin function and trafficking. We focused our attention on the interaction with: p120, β-catenin, PIPKIγ and Hakai. We showed that cytoplasmic E-cadherin mutations affect the interaction of one or more binding partners, compromising the E-cadherin stability at the plasma membrane and likely affecting the adhesion complex competence. In the present work, we demonstrated that the study of the interplay between E-cadherin and its binding partners, using PLA, is an easy, rapid, quantitative and highly reproducible technique that can be applied in routine labs to verify the pathogenicity of E-cadherin missense mutants for HDGC diagnosis, especially those located in the intracellular domain of the protein.     

Loss of E-cadherin expression in early gastric cancer

AIMS: Reduction or loss of E-cadherin expression was examined in early gastric carcinomas and precursor lesions with the following aims: (1) to assess overall E-cadherin expression in various stages of gastric carcinogenesis; (2) to correlate E-cadherin expression with the Lauren type, the grade of differentiation and the type of growth pattern of the tumours; and (3) to correlate E-cadherin expression with lymph node metastasis and overall prognosis. METHODS AND RESULTS: Forty-five paraffin-embedded gastrectomy specimens from early carcinomas were examined for the presence of various precursor lesions. The Lauren type, the grade of differentiation and the type of growth pattern were reassessed for all early carcinomas. E-cadherin expression was examined using antibody HECD-1. Whereas E-cadherin was strongly and evenly expressed in the gastric foveolar epithelium, intestinal metaplasia and early gastric carcinomas showed a lower expression. A significant difference in E-cadherin expression was found between the Lauren types (P < 0.0001). Moreover, an inverse correlation was found between E-cadherin expression and histological grade (P < 0.0001). Neither a difference in E-cadherin expression between the various growth types nor an association with lymph node metastasis and overall prognosis was found. CONCLUSIONS: The Lauren types differ in E-cadherin expression, although reduced E-cadherin expression in all probability rather reflects poor differentiation than a diffuse growth pattern 'genotype'. Moreover, E-cadherin expression does not underlie the difference in biological behaviour of early carcinomas with different types of growth pattern. Finally, E-cadherin expression is not associated with lymph node status and 5-year survival rate, at least not in early gastric carcinomas.     

The adenoma-carcinoma sequence in AMMN-induced colonic tumors of the rat

There is general agreement today that most colorectal carcinomas develop from adenomas via the so-called adenoma-carcinoma-sequence. There has however in addition been a discussion regarding the possibility of carcinomas arising "de-novo". The aim of the present study was to find out alterations preceding carcinomas in animal experiment and to demonstrate possible similarities to man. For colonic tumor induction in 40 male Sprague-Dawley rats the local acting nitrosamine-derivative acetoxymethyl-methylnitrosamine was instilled intrarectally once a week for 6 weeks in a dosage of 2 mg/kg bodyweight. 14 weeks after the end of the tumor induction those 28 animals, that had developed endoscopically recognizable tumors, were randomized into two groups. The animals from group I were sacrificed 3 weeks, the animals from group II 20 weeks later, that is 23 and 40 weeks after the start of the experiment. At histopathologic examination an increase of the number of tumors per animal and a doubling of the mean tumor diameter could be observed in group II. The most severe findings (adenoma, adenoma with dysplasia, carcinoma) were more pronounced in group II. "De-novo" carcinomas without residues of benign adenomas were not found. It could be demonstrated, that chemically induced colonic tumours produced in the way described, develop in a fashion quite similar to the conditions found in man via the adenoma-carcinoma-sequence.     

E-cadherin expression in follicular carcinoma of the thyroid

Follicular carcinoma (FC) of the thyroid is distinguished from follicular adenoma (FA) by confirmation of invasion or metastasis. However, it is still unknown how FC acquires the potential for invasion or metastasis. Twenty early FC (pT1 and pT2) were analyzed for immunohistochemical E-cadherin (E-CD) expression. Four of four (100%) widely invasive FC showed reduced E-CD expression, whereas only two of 16 minimally invasive FC showed reduced expression. In most of the minimally invasive FC, both E-CD and beta-catenin were expressed on cell-cell boundaries, even in areas where capsular invasion occurred. These results suggest that the mechanism of invasion may be different between widely invasive FC and minimally invasive FC, and that E-CD is probably responsible for the invasion of widely invasive FC. Analysis of methylation of the E-CD gene promoter using the methylation-specific polymerase chain reaction (MSP) method revealed no methylated alleles in the DNA from FC.     

Characterization of the E-cadherin/catenin complex in colorectal carcinoma cell lines

The E-cadherin/catenin complex is a prime mediator of cell-cell adhesion. APC mutations can result in loss of beta-catenin downregulation and an accumulation of beta-catenin in the cell. Beta-CATENIN mutations can have a similar effect. The aim of this study was to investigate the effect of beta-CATENIN and APC mutations on the expression and assembly of the E-cadherin/catenin complex. Five colorectal carcinoma cell lines with different APC and beta-CATENIN gene status were selected and mutations were confirmed. The expression of members of the E-cadherin/catenin complex was studied by immunohistochemistry and Western blotting. Complex assembly was investigated by immunoprecipitation. It is shown that E-cadherin and catenins are expressed in colorectal carcinoma cell lines with the predominant complex assembly being E-cadherin/beta-catenin/alpha-catenin. The subcellular distribution of the proteins is influenced by cell-cell contact, resulting in membranous localization. The expression and assembly of the E-cadherin/catenin complex does not appear to be affected by the presence of APC and or beta-CATENIN mutations.     

The ultrastructural immunohistochemistry of oncofoetal antigens in large bowel carcinomas

Summary Seven large bowel carcinomas were examined by light and electron microscopy for the presence of five oncofoetal antigens. Ultrastructural investigations involved a novel method whereby thick sections of gluteraldehyde-fixed material were cut on a vibratome and then labelled using slight modifications of a standard unlabelled antibody-enzyme (PAP) technique, before further processing.Ultrastructural preservation, staining properties and the retention of antigen activity was seemingly better than that achieved by other investigators.Specific, positive labelling for carcinoembryonic antigen (CEA), colon specific antigen (CSA) and pregnancy-specific-1-glycoprotein (SP1) was seen in every case. Clear positive labelling for placental alkaline phosphatase (PLAP) and human chorionic gonadotropin (HCG) was seen in two cases.Extracellular labelling was found in areas of cell debris, free lying or in phagocytic cells and on tumour cell brush borders. The pattern of intracellular labelling, however, was different for each antigen and reflected the probable sites of synthesis and release from the cells.Thus CEA, a complex glycoprotein, was localised within the golgi apparatus, small apical cytoplasmic vesicles and mucous droplets in relatively well differentiated tumour cells. CSA, a chemically related glycoprotein, had a similar, but less dense distribution. SP1, by contrast, was localised within basally-located vesicles associated with the ribosomal endoplasmic reticulum and appeared to be released and persist as debris or taken up by phagocytic cells below the basal lamina. PLAP and HCG, both proteins, were found within simple single membrane-bound vesicles within relatively undifferentiated cells.     

喉鳞状细胞癌中FAK和E-cadherin的表达及意义

目的 探讨粘着斑激酶(FAK)和E-钙黏蛋白(E-cadherin)在喉鳞状细胞癌(LSCC)组织中的表达与肿瘤的病理学分级及颈淋巴结转移之间的关系.方法 应用逆转录多聚酶链反应(RT-PCR)和免疫组化sP法检测LSCC和癌旁组织中FAK和E-cadherin表达情况.结果 在癌和癌旁组织中,FAK和E-cadherin mRNA的相对表达量差异具有显著性(P<0.05),且两者的表达与LSCC的淋巴结转移相关(P<0.05).E-cadherin mRNA相对表达量与肿瘤病理学分级相关(P<0.01).在癌及癌旁组织中FAK和E-cadherin的蛋白表达差异具有显著性(P<0.01),并且与颈部淋巴结转移有关(P<0.05);E-cadherin蛋白表达与肿瘤病理学分级相关(P<0.05).FAK与E-cadherin在mRNA水平或蛋白水平其表达呈负相关(r=-0.287,P<0.05;r=-0.287,P<0.05).结论 FAK的表达增高与E-cadherin的表达降低可能参与了LSCC的发生过程,是LSCC恶性程度及侵袭转移的重要标志物,对预测LSCC预后具有重要意义.     

胃癌组织中E-cadherin和claudin-4的表达及临床意义

目的:研究E-cadherin和claudin-4在胃癌组织中的表达及其临床病理意义.方法: 采用免疫组织化学方法,观察98例胃癌组织中E-cadherin 和claudin-4表达,分析其与临床病理特征的关系及各参数对预后的影响.结果: 胃癌组织E-cadherin失表达与淋巴结转移和TNM分期相关.claudin-4过表达与TNM分期相关.单因素分析显示胃癌患者预后与E-cadherin失表达、浸润深度、淋巴结转移、TNM分期相关.结论: E-cadherin失表达和claudin-4过表达与胃癌的发生进展有关,E-cadherin失表达影响胃癌患者预后.