c-myc, c-erbB-1 and c-erbB-2 expressions in urothelial carcinoma

The expression of c -myc , c- erb B-1 and c- erb B-2 In 24 cases of urothelial carcinoma by Southern and northern blot analysis, and immunohistochemistry was examined. The results were compared with the pathological grade and stage. We found elevated mRNA expressions of c- myc and c- erb B-1 In 19 and 11 of 21 cases, respectively, but there was no apparent amplification or rearrangement of these oncogenes in any of the cases examined. By immunohistochemistry using anti-epidermal growth factor receptor antibody, most of the cases showed positbe immunoreactivity on the cancer cell membranes, and cancers of higher pathological grade and stage showed more intense staining. By contrast, amplification of c- erb B-2 was detected in four of 24 cases, all of which were assigned to a high pathological grade (G3). Elevated c- erb B-2 mRNA levels appeared to correlate with the pathological grade of the cancers. Positive immunohistochemical reactions to c- erb B-2 were found in the cancer cell membranes in three of 24 cases, which were accompanied by amplification and eievated mRNA levels of c -erb B -2 . In conclusion, expressions of c- myc , c- erb B-1 and c- erb B-2 were all elevated in the majority of urothellal carcinomas, but the amplification was not universal.     

The c-erbB-2 Protein in Primary and Metastatic Breast Carcinomas

Material from 41 patients with primary breast carcinoma and lymph node metastases at the time of primary surgical intervention was immunostained for c-erbB-2 protein, neuron-specific enolase (NSE), and estrogen receptors. Thirty of the primary breast carcinomas were of ductal type. Six were classified as infiltrating lobular carcinomas, 2 were apocrine, 1 was mucinous, and 1 was a tubular carcinoma. One tumor could not be classified as ductal or lobular by light microscopic examination alone. The number of lymph node metastases available varied from 1 to 14 per case (median, 3.9). Nine (22%) of the primary breast carcinomas (8 ductal and 1 apocrine) expressed c-erbB-2 protein and showed c-erbB-2 gene amplification; 12 expressed NSE immunoreactivity. None expressed both markers. Estrogen receptor immunoreactivity was present in 23 of the 41 cases, including 9 of the NSE-positive cases. C-erbB-2 protein-positive metastases were present in 18 cases (44%), and in 13 cases all metastases were immunostained. In 5 cases the expression of c-erbB-2 protein varied from metastasis to metastasis. NSE immunoreactivity was expressed in 10 cases, and in 3 cases with minor NSE-positive cell populations the metastatic lesions expressed c-erbB-2 protein as well. All 9 primary breast carcinomas expressing c-erbB-2 protein had lymph node metastases with c-erbB-2-immunoreactive tumor cells. Eight of the 9 c-erbB-2 protein-negative primary tumors with metastases expressing c-erbB-2 protein showed no amplification of the c-erbB-2 gene. Thus expression of c-erbB-2 protein can occur during the metastatic process, even if it seems to be missing in the primary tumor. On the other hand, if a primary breast carcinoma expresses c-erbB-2 protein, this feature seems to be present in all the tumor metastases as well.     

Immunohistochemical distribution of c-erbB-2 in in situ breast carcinoma--a detailed morphological analysis

An immunohistochemical study of c-erbB-2 expression was carried out on in situ (non-invasive) breast carcinoma, using antibody 21N, raised to the intracytoplasmic domain of the c-erbB-2 oncogene product. Strong membrane staining was observed in 44 out of 74 (59 per cent) cases of ductal carcinoma in situ (DCIS), but none of 48 lobular carcinoma in situ (LCIS) lesions. A detailed comparative morphological evaluation using several different parameters, including histological subtypes, was performed within the DCIS group. The results showed that there was a significant correlation between c-erbB-2 expression and the presence of large cell size, periductal lymphoid cell infiltration, marked nuclear pleomorphism, multinucleation, and a high mitotic rate. Of these, cell size appears to be the most important predictor of c-erbB-2 status, followed by the presence of periductal lymphoid cell infiltration. These results indicate, firstly, that LCIS and DCIS are biologically (as well as histologically) different and, secondly, that a subgroup of DCIS, which is associated with c-erbB-2 over-expression, exists and appears to have distinct histological features. The subgroup of DCIS cases which over-express c-erbB-2 may be a biologically definable category with prognostic importance. These results may therefore have relevance to breast screening programmes, but a larger study incorporating clinical data would be necessary to correlate these findings with clinical outcome.     

乳腺癌钼靶X线显示钙化灶与ER、PR和c-erbB-2表达的相关性研究

目的:探讨乳腺癌钼靶X线钙化与ER、PR及c-erbB-2表达的相关性及其临床意义.方法:分析86例乳腺癌钼靶X线片,将其分为有钙化组(46例)和无钙化组(40例);将切除的肿瘤标本行HE及免疫组织化学染色,检测并比较两组ER、PR和c-erbB-2的表达率.结果:乳腺癌钼靶X线有钙化组ER和PR的表达率均略高于无钙化组(56.5% vs 55.0%,45.6% vs 42.5% ),但差异无统计学意义 (P＞0. 05);有钙化组c-erbB-2表达率高于无钙化组(65.2% vs 40.0%),差异有统计学意义(P＜0. 05).结论:乳腺癌钼靶X线钙化与ER、PR表达无相关性,但与c-erbB-2表达密切相关;钼靶X线钙化可以粗略反映c-erbB-2表达情况,可为乳腺癌治疗策略的制定和预测预后提供参考.     

Aberrant FGF-2, FGF-3, FGF-4 and C-ERB-B2 Gene Copy Number in Human Ovarian,Breast and Endometrial Tumours

Abstract

The important role of oncogene amplification and tumour suppressor gene deletion in human tumours is becoming increasingly apparent. However, extensive screening of human tumours is required before the prognostic significance of such genetic abnormalities can be fully appreciated. The present investigation describes a rapid non-radioactive and largely automated procedure for the analysis of aberrant gene copy number in large numbers of tissue samples of different human rumours. This procedure is based on the sequential use of the polymerase chain reaction (PCR) and high performance ion exchange liquid chromatography (HPIEX). Using this rapid PCR/HPIEX technique, we have identified amplification and deletion of the FGF-2 gene and the FGF-3, FGF-4 and c-erb-B2 oncogenes in human tumours of the breast, ovary and endometrium. Comparison of the data with tumour pathology has revealed possible associations between aberrant gene copy number and tumour type, invasiveness and metastases.     

Detection of c-erbB-2 (HER-2/neu) amplification in breast carcinoma by fluorescence in situ hybridization on tissue sections and imprinted cells

Abnormalities in c-erbB-2 have attracted a great deal of attention. Treatment using an antibody against the c-erbB-2 gene product is effective against breast cancers with amplification and/or overexpression of c-erbB-2. There is an urgent need to establish methodology for selecting patients who would benefit from this therapy. A total of 235 breast carcinomas were examined for c-erbB-2 protein overexpression by immunohistochemistry. Tissue sections with discernible immunostaining from 52 tumors, 70 negative tumors and smear imprints from 35 patients were examined by dual-color fluorescence in situ hybridization (FISH) using probes specific for c-erbB-2 and the centromeric region of chromosome 17. The concordance between gene amplification and protein overexpression was 95.7%. When the findings of the two FISH preparation techniques were compared, no discrepancies were found in 24 of the tumors. However, differences were seen in eight cases. In six of these cases the differences did not affect the presence or absence of amplification, but in the other two cases, considered to show low-level amplification on paraffin sections, polysomy 17 was detected instead. It was concluded that FISH is an excellent tool to detect gene amplification in particular, and FISH on touch imprints is a useful adjunct to differentiate between low-level amplification and polysomy 17.     

c-erbB-2 oncoprotein expression in mammary and extramammary Paget''s disease: an immunohistochemical study

Over-expression of the c-erbB-2 oncogene occurs in a proportion of human adenocarcinomas and in breast carcinoma is associated with poorer prognosis. Sections of formalin-fixed, paraffin-embedded tumour tissue from 22 patients with mammary and extramammary Paget's disease have been stained immunohistochemically using a monoclonal antibody (NCL-CB11) raised against a synthetic peptide from the C-terminal end of the predicted sequence of the c-erbB-2 protein product. All 12 cases of mammary Paget's disease showed membrane staining of intra-epidermal cells, indicating c-erbB-2 over-expression. Sections of underlying ductal breast carcinoma were available in nine cases; all nine tumours were c-erbB-2 positive and in eight the in situ component was of comedo or solid type. There was membrane staining of tumour cells in four of the 10 cases of extramammary Paget's disease; staining intensity was generally weaker than that observed in the cases of mammary disease. The possible implications of these findings for the histogenesis of both mammary and extramammary Paget's disease are discussed.     

Overexpression of p53, c-erbB-2 and epidermal growth factor receptor in human breast carcinomas

Overexpression of p53 protein, epidermal growth factor receptor (EGF-R), and c-erbB-2 protein was assessed by immunohistochemical staining of formalin-fixed, paraffin-embedded tissue from 64 invasive breast tumors. The correlation between abnormal expression of each protein and various disease parameters, including lymph node metastasis and histopathologic type and grade was analyzed. Despite the previous proposal, no significant correlation was found between lymph node metastases and overexpression of each gene in the primary tumors. In addition, some metastatic lesions did not always exhibit overexpression, even if it was evident in the primary tumors. Overexpression of c-erbB-2 protein correlated well with Bloom's histological grading. p53 expression was detected most often in tumors with hyperchromatism and more frequent mitosis. Overexpression of c-erbB-2 protein occurred more frequently in p53-positive tumors. The results indicate that abnormal expression of p53 protein causes genetic instability in the early stage of tumor development, resulting in subsequent overexpression of other oncogenes.     

Over-expression of c-erbB-2 correlates with nuclear morphometry and prognosis in breast carcinoma in Asian women

AIM: We aimed to investigate the immunohistochemical expression of c-erbB-2 in invasive breast carcinoma in Asian women and its correlations with clinicopathological parameters and nuclear morphometry. Patients were followed up for disease relapse and overall survival, and the data were reviewed in conjunction with c-erbB-2 over-expression. METHODS: Paraffin sections from 321 invasive breast cancers were immunohistochemically stained with anti-human c-erbB-2 antibody using the streptavidin-biotin technique. RESULTS: c-erbB-2 was over-expressed in 110 (34.3%) cases, with an inverse correlation with oestrogen receptor (ER) and progesterone receptor (PR) status (p=0.0001) and a positive correlation with histological grade (p=0.017). Nuclear morphometry in 96 cases revealed rounder nuclei in c-erbB-2 negative tumours (p=0.0322) when compared with c-erbB-2 positive tumours. Among c-erbB-2 positive cases, malignant cells of histological grade 3 tumours revealed larger nuclear area and perimeter than grade 1 and 2 cases (p=0.0095, p=0.03, respectively) while increasing tumour size correlated with greater nuclear perimeter (p=0.046). c-erbB-2 positivity was significantly associated with poor survival when all patients were included in the analysis (p=0.0166) and for subsets of node positive, histological grade 1 and 2, and ER positive tumours, and in women aged over 50 years (p=0.0047, p=0.0367, p=0.0092, p=0.0096, respectively). CONCLUSIONS: c-erbB-2 was independently prognostic when histological grade, nodal and ER status were considered. Our results show that c-erbB-2 over-expression correlates with poor histological grade and negative ER/PR status, and predicts poor overall survival in Asian women with breast cancer.     

c-erbB-2 expression in FNAB smears and matched surgical specimens of breast cancer

Most of the data regarding the significance of c-erbB-2 oncogene expression as a prognostic marker in breast cancer have been generated in many large retrospective studies by retrieving the corresponding oncoprotein in archival paraffin embedded sections. Recently, employing fresh breast cancer cells obtained by means of fine-needle aspiration biopsy, we found a rate of c-erbB-2 positive breast tumors (58%) higher than that reported in paraffin-embedded tissue sections by others studies. The present analysis was undertaken to investigate the impact of routine tissue processing on the preservation of the c-erbB-2 immunoreactivity. This issue was addressed by assessing the relative rate of c-erbB-2 oncoprotein immunodetection on FNAB smears and matched surgical specimens of breast cancer. The expression of c-erbB-2 oncoprotein was evaluated using the alkaline phosphate-anti-alkaline phosphatase (APAAP) technique in 54 breast aspirates and corresponding surgical specimens of primary breast cancer. Twenty-six (48%) smears and 23 (43%) matched paraffin sections gave specific signal for c-erbB-2 oncoprotein. The slightly higher incidence of c-erbB-2 expression found on smears seems to be mainly due to the better antigen preservation in the fresh cytological preparations. We conclude that routine histological processing may affect c-erbB-2 immunoreactivity; therefore, in mounting prospective studies, it is advisable to assess c-erbB-2 status in fresh tissue. Moreover, the assessment of c-erbB-2 expression on aspirate samples may yield additional information to the pre-surgical prognostic evaluation of breast cancer diagnosed by FNAB. Diagn Cytopathol 1996;14:135–139. © 1996 Wiley-Liss, Inc.     

Absence of microsatellite instability in breast carcinomas with both p53 and c-erbB-2 alterations

Based on a previous finding that amplification of the c-erbB-2 oncogene and alteration of p53 are strongly associated in most aggressive breast tumours, the present study investigated whether microsatellite instability (MI) might also be associated with this tumour phenotype. Nine polymorphic microsatellite markers, including six dinucleotide, one trinucleotide, and two tetranucleotide repeats, were amplified from paired normal and tumour DNA samples of 15 breast tumours that overexpressed both c-erbB-2 and p53 and of 15 control breast tumours that overexpressed neither protein. All 30 breast tumours analysed exhibited a replication error-negative phenotype, with only one sample showing MI in one of the nine loci. This suggests that the genetic events underlying MI, which are critical in colorectal and gastric tumours, are not involved in the pathogenesis of c-erbB-2/p53 double-altered breast tumours and do not play a central role in breast tumour formation. Copyright © 1999 John Wiley & Sons, Ltd.     

Immunocytochemical localization of c-erbB-2 protein in transitional cell carcinoma of the urinary bladder

The level of expression and cellular localization of the c-erbB-2 gene product in transitional cell carcinoma of the urinary tract is controversial. Analysis of the c-erbB-2 gene structure and comparison of its expression in the same cells by Southern, Northern and immunoblotting, and by immunocytochemistry minimize the errors of interpretation inherent in one technique. Such a ‘correlative study’ has been performed on tumours from 82 patients, c-erbB-2 gene amplification was detected in 14 per cent of initial tumours and was associated with grade (.P< 0·001). Raised levels of mRNA were seen in those tumours with increased gene copy number and in 13 per cent of the remainder. Immunoblotting detected the expected 185 kD immunoreactive protein and a 155 kD piotein associated with high gene copy number. Immunocytochemistry localized c-erbB-2 immunoreactivity to the cell membrane and cytoplasm, and the latter predominated. Four antibodies to c-erbB-2 (AB-3, 21N, pAb 1, and NCL CB11) were compared on contiguous sections of the same tumour and showed the same pattern of immunoreactivity. Similarly, analyses carried out in three independent laboratories identified the same cellular localization. Membrane and cytoplasmic immunoreactivity was demonstrated in all tumours with gene amplification or increased mRNA levels and in 40 per cent of the remaining tumours. We showed that immunocytochemistry requires careful standardization of techniques and quantitation between different groups. However, despite variations in the intensity of immunoreactivity, the total number of positive cells remained constant. Therefore quantitation must be based on the number of positive cells and, ideally, their immunoreactive content relative to normal and positive tissue controls.     

Cytoplasmic c-erbB-2 protein expression correlates with survival in Dukes' B colorectal carcinoma

The prognostic significance of c- erb B-2 expression was studied in paraffin wax embedded colorectal cancer tissue using a monoclonal antibody. One hundred and sixty-four patients with Dukes' B disease were studied. Membranous staining was not detected in any case. Cytoplasmic c- erb B-2 staining was seen in 55 cancers (33.5%). Cytoplasmic taining was unrelated to patient age ( P = 0.31), sex ( P = 0.69), tumour site ( P = 0.69), size ( P = 0.57), histological grade ( P = 0.42) or ploidy status ( P = 0.21) but was found more frequently in obstructing cancers ( P = 0.03). Mean follow up of the patient population was 6.3 years. Five-year-survival estimated by the Kaplan-Meier life-table method was 47% for those with cytoplasmic c- erb B-2 staining and 77% for those without (log rank analysis; P ⋜ 0.0001). Stepwise regression analysis identified c- erb B-2 staining (relative risk, 2.51; P = 0.0005) and bowel obstruction (relative risk, 1.99; P = 0.015) as independent predictors of survival. It is suggested that cytoplasmic c- erb B-2 expression may provide a useful marker of tumour behaviour in Dukes' B colorectal cancer.     

环氧化酶-2、c-erbB-2和Ki67在结直肠癌中表达及其临床意义

目的：探讨环氧化酶-2(COX-2)、c-erbB-2、Ki67在结直肠癌中的表达,相互关系及其临床意义。方法：采用免疫组化S-P法检测结直肠癌中三者的表达。结果：结直肠癌组中,(1)COX-2、c-erbB-2蛋白阳性表达与淋巴结转移及Duckes分期有关;(2)COX-2、c-erbB-2、Ki67蛋白表达水平,两两间皆呈正相关;(3)三者联合分析显示：COX-2( )c-erbB-2( )组的Ki67平均标记指数显著高于单独COX-2或c-erbB-2阳性,或共同阴性组Ki67的表达。结论：COX-2、c-erbB-2及Ki67在结直肠癌发生发展中起着重要作用,三者联合检测有助于结直肠癌的细胞增殖活性,恶性程度的判断。     

KiSS-1 Expression in Human Breast Cancer

The KiSS-1 gene encodes a 145 amino acid residue peptide that is further processed to a final peptide, metastin, a ligand to a G-coupled orphan receptor (OT7T175/AXOR12). KiSS-1 has been identified as a putative human metastasis suppressor gene in melanomas and in breast cancer cell lines. This study aimed to determine the expression and distribution of KiSS-1 and its receptor in human breast cancer tissues and to identify a possible link between expression levels and patient prognosis. Frozen sections from breast cancer primary tumours (matched tumour 124 and background 33) were immuno-stained with KiSS-1 antibody. RNA was reverse transcribed and analyzed by Q-PCR (standardized using β-actin, and normalized with cytokeratin-19 levels). Levels of expression of KiSS-1 were higher in tumour compared to background tissues (3124±1262 vs 2397±1181) and significantly increased in node positive tumours compared to node negative (3637±1719 vs 2653±1994, P = 0.02). KiSS-1 expression was also increased with increasing grade and TNM status. There were no such trends with the KiSS-1 receptor. Expression of KiSS-1 was higher in patients who had died from breast cancer than those who had remained healthy (4631±3024 vs 2280±1403) whereas expression of the receptor was reduced (480±162 vs 195±134). Immunohistochemical staining showed increased expression of KiSS-1 in tumour sections. Insertion of the KiSS-1 gene into the human breast cancer cell line MDA-MB-231, resulted in cells that were significantly more motile and invasive in behaviour, with reduced adhesion to matrix, using respective assays. In conclusion, KiSS-1 expression is increased in human breast cancer, particularly in patients with aggressive tumours and with mortality. Over-expression of KiSS-1 in breast cancer cells result in more aggressive phenotype. Together, it suggests that KiSS-1 plays a role beyond the initial metastasis repressor in this cancer type.     

c─erbB─2、AgNOR在乳腺癌中的预后价值及其相互关系探讨

对８８例乳腺癌作了ＡｇＮＯＲ银染计数及ｃ─ｅｒｂＢ─２癌蛋白的免疫组化研究。结果发现ｃ─ｅｒｂＢ─２阳性者，病死率大于ｃ─ｅｒｂＢ─２阴性者（Ｐ＜０．０１）；ＡｇＮＯＲ计数＞５者，病死率大于ＡｇＮｏＲ≤５者（Ｐ＜０．０１），表明了这两个指标对预后具有明显的价值。同时发现ＡｇＮＯＲ值＞４者，ｃ─ｅｒｂＢ─２阳性率显著高于ＡｇＮＯＲ≤４者（Ｐ＜０．０５），表明这两个指标之间也有一定的关系。我们还发现ｃ─ｅｒｂＢ─２阳性、ＡｇＮＯＲ值＞５者，病死率显著大于ｃ─ｅｒｂＢ─２阴性、ＡｇＮＯＲ值≤５者组。     

Multiple developmental pathways of highly aggressive breast cancers disclosed by comparison of histological grades and c-erbB-2 expression patterns in both the non-invasive and invasive portions

To determine the developmental stages at which the highly malignant phenotype of breast carcinoma is acquired, the histological grade and c- erbB -2 oncoprotein expression status were examined for both the ductal carcinoma in situ (DCIS) and Invasive components of 437 separate Invasive breast carcinomas. In 218 invasive carcinomas with high-grade atypla (grade 3), the DCIS components were grade 2–3 In 158 cases (73%). Twenty-seven (12%) showed an obvious stepwise Increase from grade 1 DCIS to grade 3 invasive carcinoma, and 25 of these tumors had DCIS components covering > 325% of their area. Ductal carcinoma in situ components were undetectable In 33 (15%) of invasive carcinomas. The Incidence of c-erb B -2 overexpression was higher in grade 3 carcinomas with grade 2–3 DCIS components (55%, 80 of 146) than in those with grade 1 DCIS components (5%, one of 25). The Incidence was also higher in grade 3 carcinomas with DCIS components covering ≥ 25% of the tumor area (71%, 39 of 55) than in those with DCIS over > 325% of the total area (36%, 42 of 116) or without DCIS components (6%, two of 33). There appeared to be three prototypic pathways to high-grade breast carcinoma: (i) invasion by a high-grade DCIS regardless of the extent of DCIS spread; (II) invasion by a low-grade DCIS during the microscopic stages, accompanied by an obvious enhancement of the grade; and (iii) development of an invasive carcinoma ab inltlo. c-erb B-2 overexpression appeared to be frequently involved in the early development of the first group but showed little relation to the invasive process in any of these three pathways. The histological grade and c- ert B-2 overexpression appeared to be largely established during the early microscopic stages of a DCIS or invasive carcinoma.     

乳腺癌中p27和c-erbB-2的表达及意义

目的　探讨乳腺癌中 p2 7和c erbB 2表达情况和意义。 方法　用免疫组化S P法检测 4 0例乳腺癌中 p2 7和c erbB 2的表达。结果　正常乳腺组织c erbB 2表达阴性 ,癌组织中阳性率为 37 5 % (15 / 4 0 ) ,两者差异有显著性 (P <0 0 0 1)。 4 0例乳腺癌组织中 p2 7高表达率为 32 5 % (13/ 4 0 ) ,正常乳腺组织高表达为 80 % ,两者差异有显著性 (P <0 0 0 1)。p2 7高表达与癌组织分化、淋巴结转移和复发有关 (P <0 0 5 )。结论　基因p2 7和c erbB 2表达异常是乳腺癌产生的机制之一 ,与乳腺癌的发生及发展有关。p2 7可能是乳腺癌的一个重要预后指标     

Concordance in judgments among c-erbB-2 (HER2/neu) overexpression detected by two immunohistochemical tests and gene amplification detected by Southern blot hybridization in breast carcinoma

We compared the c-erbB-2 protein overexpression status detected by the HercepTestTM (DAKO A/S, Grostrup, Denmark) with another conventional immunohistochemistry system using an anti-c-erbB-2 rabbit polyclonal antibody (Nichirei Co., Tokyo, Japan) and with the c-erbB-2 gene amplification status detected by Southern blot hybridization in 101 surgically resected breast carcinomas. According to the criteria for overexpression, recommended by the manufacturer, c-erbB-2 overexpression by HercepTestTM was detected in 24 cancers (24%), comprising six score-2 tumors and 18 score-3 tumors. The level of agreement in judgment of the HercepTestTM, among three independent observers, was excellent (kappa = 0.845). C-erbB-2 overexpression by Nichirei's antibody and c-erbB-2 gene amplification were detected in 21% and 16% of cases, respectively, and their concordance with HercepTestTM scores of 2-3 was 89% and 90%, respectively. In the score-3 cases by Hercep TestTM only, the concordance rates with overexpression by Nichirei's immunohistochemistry and with gene amplification were slightly higher, 94% and 93%, respectively. Score-2 cases by HercepTestTM were mostly judged as negative overexpression by Nichirei's antibody and as no amplification by Southern blot hybridization. The present results showed that HercepTestTM score-3 detected c-erbB-2 overexpression almost optimally as well as the conventional methods and the score-3 breast carcinomas had clinical and biological implications. Further examination would be necessary to decide the significance of breast cancers of HercepTestTM score 2.