Identification and characterization of c-erbb-2 proteins in serum,breast tumor tissue,and sk-br-3 cell line

We have identified and characterized c-erbB-2 protein molecules in sera from patients with carcinomas, in both cytosol and cell membrane extract from breast tumor tissue and in both the culture medium and cell extract of the SK-BR-3 cell line. These proteins were characterized by various chromatographic techniques and identified by the use of two immunoassays; one measures both the c-erbB-2 oncoprotein (p 185) and its ectodomain (p 120), and the other in-house assay reacts specifically for p 185. We found that the majority of the immunoreactivity detected in the serum, tumor tissue cytosol, and conditioned cell medium was derived from the ectodomain molecule (p 120) of the c-erbB-2 oncoprotein (p 185), whereas only p185 was detected in the extracts from cell membrane of both tumor tissue and the SK-BR-3 cell line. The ectodomain molecules (p 120) found in the serum, cytosol, and cell medium were very similar in terms of molecular size and charge property. The molecular weight was determined to be 120 kDa by the size exclusion HPLC method. Both p 120 and p 185 are glycoproteins and were retained by the ConA Sepharose column. Both molecules are also heterogeneous in charge and multiple peaks could be identified in the elution profiles of anion exchange HPLC and chromatofocusing. This information should not only facilitate the isolation of these molecules, but also improve preparation of specific antibodies, preparation of calibrators, and development of improved assays for these proteins.©1995 wiley-Liss, inc.     

Measurement of c-erbb-2 proteins in sera from patients with carcinomas and in breast tumor tissue cytosols: Correlation with serum tumor markers and membrane-bound oncoprotein

Using a commercial kit with antibodies against the ectodomain of c-erbB-2 protein, we detected c-erbB-2 immunoreactivity in human serum. We found that the percentages of patients with elevated serum c-erbB-2 immunoreactivities were 35, 21, and 9% in breast, prostate, and ovarian carcinoma, respectively. The majority of the elevated immunoreactivities were associated with sera containing highly elevated tumor markers with the highest in breast carcinoma (35%) and lowest in ovarian cancer (9%). Excellent correlations were also observed between the serum levels of c-erbB-2 immunoreactivity and the dominant tumor markers in serial specimens from individual cancer patients. We could also detect the c-erbB-2 immunoreactivity in the cytosols prepared from the breast tumor tissue for estrogen and progesterone receptor (ER&PgR) measurements using the same commercial kit for serum studies, and the intact c-erbB-2 oncoprotein (p 185) in the extracts of the tissue membrane fractions with a different kit designed for tissue extract. The level of c-erbB-2 immunoreactivity in the cytosol from 124 human breast tumor specimens had an excellent correlation with the cell membrane concentrations of p 185 (γ = 0.89). Most of the elevated cytosol c-erbB-2 immunoreactivities were also found to associate with breast tumor specimens containing low concentrations of ER&PgR. It appears that measuring the c-erbB-2 immunoreactivity potentially could be used as a prognostic marker without performing tissue biopsies and also as a serum tumor marker for managing cancer patients.©1995 wiley-Liss, inc.     

Isolation of the intact molecule and ectodomain of C-erbB-2 oncoprotein from SK-BR-3 cells and development of immunoassays on microplate

We isolated both the intact molecule (p185) and the ectodomain (p120) of c-erbB-2 oncoprotein from SK-BR-3 breast tumor cells. The p120 was extracted from the cells by 0.05 M phosphate buffer, pH 7.2, whereas the extraction of the p185 required the presence of a detergent, such as 1% Triton X-100 in 0.05 M Tris buffer. Protease inhibitors were also included in the extraction buffer during the isolation of p185 in order to prevent cleavage of p185 to p120 by an unknown protease apparently also present in the extract. In case there was any p120 in the p185 preparation, the p120 could be separated from p185 by chromatography on a Superose 12 column. Using the p120 and p185 as calibrators, we have established two microplate sandwich immunoassays: one measures both p185 and p120 (total assay) and the other is specific for the p185. Since capturing and detecting antibodies used in the total assay react against the extracellular domain of the c-erbB-2 oncoprotein, they can therefore be used to measure the p120 in serum and p185 in breast tumor tissue cytosol. On the other hand, the p185 specific assay uses the capturing antibody against the cytosolic domain of the oncoprotein and consequently can only measure p185 in breast tumor tissue cytosol. J. Clin. Lab. Anal. 12:298–303, 1998. © 1998 Wiley-Liss, Inc.     

胃癌p53、c-erbB-2、p21、nm23基因表达的分布特点及其临床意义

目的为探讨手术切除胃癌组织p53、c-erbB-2、p21、nm23联合基因产物表达检测对胃癌诊断治疗及预后判断的价值。方法应用免疫组化技术检测了手术切除胃癌组织p53c、-erbB-2、p21、nm23基因产物表达。结果p53蛋白表达阳性率为36.4%~46.7%,c-erbB-2为33.3%~56.8%,p21为35.1%~56.7%,nm23为30.0%~70.5%。在非胃癌组织中(胃,十二指肠溃疡,胃息肉,重度不典型增生)未见c-erbB-2、p21、nm23基因表达。p53c、-erbB-2、p21的表达与胃癌的分化程度,肿瘤浸润程度,肿瘤转移程度及临床分期呈正相关。nm23的表达与胃癌的分化程度,肿瘤浸润程度,肿瘤转移程度及临床分期呈负相关。p53c、-erbB-2、p21n、m23四种肿瘤蛋白在胃镜活检标本和手术切除标本中表达无显著性差异,胃镜活检标本和手术切除标本检测阳性符合率分别为90.2%、91.0%、93.7%和89.3%。结论对胃癌组织p53、c-erbB-2、p21、nm23基因表达检测在胃部良恶性肿瘤鉴别,非手术临床分期的判断及指导临床治疗等方面具有一定价值。多基因表达检测对胃癌预后判断明显优于单基因表达检测。     

Detection of urokinase plasminogen activator receptor and c-erbB-2 in sera of patients with breast and ovarian carcinoma

The role of urokinase plasminogen activator receptor (uPAR) and c-erbB-2 in breast and ovarian cancer was investigated. Eighty patients of breast and ovarian cancer and benign lesions, as well as twenty normal controls were evaluated for the expression of c-erbB-2 by Western blotting and uPAR levels by ELISA. The c-erbB-2 and uPAR showed a significant increase in both types of cancer investigated compared to normal control and benign lesions. The frequency of c-erbB-2 was significantly higher in breast cancer lesions (p < 0.01). Levels of CA15.3 in breast cancer and CA125 in ovarian cancer were significantly higher in cases expressing c-erbB-2 (p < 0.01) than in negative c-erbB-2 cases. The uPAR showed a significant positive correlation with advanced stages of breast cancer (r = 0.7971) and ovarian cancer (r = 0.83662), while significant correlations were found for CA15.3 in breast cancer (r = 0.64967) and CA125 in ovarian cancer (r = 0.83996). Taken together, our data suggest that the c-erbB-2 and uPAR in the sera of ovarian and breast cancer act as valuable markers for the evaluation of the patients preoperatively.     

Polyclonal antibodies against gp185HER2 peptides: their putative role in the identification of a particular HER2 status in patients with breast cancer

The HER2 oncogene and its relative oncoprotein, gp185HER2, a transmembrane glycoprotein belonging to the epidermal growth factor receptor family, are overexpressed in a wide range of solid tumors including breast and ovarian cancer. In patients with breast cancer, both humoral and cell-mediated HER2 immune responses have been found as well as in some patients with gp185HER2 nonoverexpressing tumors. To establish whether peptide sequences identified as HLA-A2-restricted T-cell epitopes are expressed in breast tumor cell lines and tissues, we produced and characterized by different methodologic approaches polyclonal antibodies raised against four gp185HER2 peptides. Two of the antibodies recognized peptides eluted from the HLA-A2 groove of the mDAmB231 breast cancer cell line expressing a basal level of gp185HER2. Paraffin-embedded primary and metastatic breast tumors were specifically immunostained by all four reagents, thereby showing an overlapping reactivity. When this immunoreactivity was compared with that obtained using two different monoclonal antibodies, in 105 breast primary tumors and 36 corresponding lymph node metastases, we identified a subset of tumors that were negative with anti-gp185HER2 monoclonal antibodies and positive with the four antipeptide antibodies. Our novel observations provide in vivo evidence of the complexity involved in evaluating HER2 expression, and open a new path for understanding the biologic significance of HER2 status in breast tumors.     

Circulating levels of HER-2/neu oncoprotein in breast cancer

HER-2/neu, also known as c-erbB-2/neu, is an oncogene located in chromosome 17 which encodes HER-2/neu, a transmembrane protein belonging to the EGFR family. The external domain of this protein is released by the cell and can be studied in serum by immunoassay. HER-2/neu in serum is a specific tumor marker and only slight elevations may be found in the absence of malignancy, mainly in association with liver diseases. Likewise, the highest concentrations of this oncoprotein are found in patients with breast cancer, but lower concentrations may be found in other malignancies, particularly ovarian, prostate and lung cancer (mainly adenocarcinomas). HER-2/neu assay sensitivity in patients with untreated primary loco-regional breast cancer is <10% and seems to be related to overexpression in tissue as well as to the most important prognostic factors: tumor size and nodal involvement. Serial HER-2/neu determinations after surgery seem to be useful in the early diagnosis of recurrence, mainly in patients with HER-2/neu overexpression in tissue, but additional studies are necessary to confirm these results. HER-2/neu sensitivity (proportion of patients with abnormal values) in patients with metastasis is around 40%-45%, with a clear relationship to tissue overexpression and to site (higher in visceral metastases) and number of metastases. The clinical utility of HER-2/neu in patients with advanced disease is mainly for therapeutic monitoring. Likewise, in most of the studies published, a relationship has been found between serum HER-2/neu levels (either pretreatment or at follow-up) with tumor response.     

The clinical significance between activation of nuclear factor kappa B transcription factor and overexpression of HER-2/neu oncoprotein in Taiwanese patients with breast cancer

BACKGROUND: This study investigated the role of nuclear factor-kappa B (NF-kappaB) activity in human breast cancer with overexpression of HER-2/neu oncoprotein, as well as its role on expression of different histological grades of cancer cells taken from Taiwanese breast cancer patients. MATERIALS AND METHODS: Specimens were collected from 82 female breast cancer patients. The HER-2/neu oncoprotein was measured by immunohistochemistry. NF-kappaB activity expression was assessed by the electrophoretic mobility shift assay, and confirmed by the supershift technique using anti-P65 antibody in both breast cancer tissue and the adjacent normal tissue. The histological grades were measured by Modified Bloom-Richardson Grading Scheme. RESULTS: Of the 82 cancer specimens, 81 (98.7%) showed higher or equal expressions of NF-kappaB activity when compared to the adjacent normal tissue. Fifty-five cases (67.1%) had higher levels of NF-kappaB activity in the cancerous tissue than in the adjacent normal tissue (p<0.005). With regard to tumor size, steroid receptors, stages, histological types, and node status, there were no statistically significant differences in NF-kappaB activity between cancerous tissues and adjacent normal tissues. However, significantly higher expressions of NF-kappaB activity were seen in those cases with positive HER2/neu oncoprotein, poorly differentiated histological grades, high nuclear pleomorphisms, and high mitotic counts (p<0.05). Positive HER-2/neu overexpression of oncoprotein had higher NF-kappaB activity (86%) than negative overexpression (60%) (p<0.05). It has been shown that the NF-kappaB activity increases in the HER-2/neu oncoprotein overexpression in human breast cancer. CONCLUSION: Overexpression of HER-2/neu gene could induce NF-kappaB activity in human breast cancer cells, as has been confirmed in other research on cell lines.     

In situ distribution of oncogene products and growth factor receptors in breast carcinoma: c-erbB-2 oncoprotein, EGFr, and PDGFr-beta-subunit

An increasing body of evidence suggests that breast tumour growth is mediated by oncogene products and growth factors which are or which act through cell surface receptors. The aims of the present study were to determine how three of these receptors, c-erbB-2 protein, epidermal growth factor receptor (EGFr) and the beta-subunit of platelet-derived growth factor receptor (PDGFr-beta-subunit), can effectively be demonstrated by immunohistochemical methods in breast tumors, how these receptors are distributed at the cellular level and how their expression correlates with well-established prognostic indicators including hormone receptors and proliferative index. We examined frozen tissue sections of 50 invasive human breast carcinomas, including 45 ductal, four lobular, and one mucinous tumours, by immunocytochemical methods to determine the in situ distributions of c-erbB-2, EGFr, and PDGFr-beta-subunit. We compared staining for c-erbB-2 protein in frozen sections with that in paraffin sections of the same 50 tumours. The immunohistochemical labelling results were compared with tissue hormone receptor content and growth fraction determined by Ki-67 labelling. Strong labelling of tumour cells in frozen sections was detected in 22% of cases, all of the ductal type, stained with rabbit antiserum to c-erbB-2. Labelling for c-erbB-2 protein was generally weaker in paraffin sections than in frozen sections and in six of 11 positive cases, specific staining could be detected only in frozen sections. In immunostains with monoclonal antibody to EGFr, rare cells within tumour were labelled in 60% of the carcinomas. Using a monoclonal antibody to the beta-subunit of PDGFr, consistent labelling of fibrillary cellular processes in the walls of blood vessels and in fibrous stroma around tumour cell nests was detected, but there was no labelling of tumour cells themselves. C-erbB-2 oncoprotein positive tumours were found to be more often oestrogen receptor negative (P less than 0.005) or oestrogen and progesterone receptor negative (P less than 0.01) than c-erbB-2 negative tumours. No significant correlation was observed between c-erbB-2 expression and Ki-67 growth fraction.     

荧光原位杂交和基因组半定量法检测乳腺癌组织中c-erbB-2基因扩增

目的利用荧光原位杂交(FISH)与基因组半定量法检测乳腺正常良性和恶性组织中原癌基因c-erbB-2的扩增情况。方法收集50例临床切除的乳腺肿瘤组织,抽提收集样品基因组DNA并扩增c-erbB-2的特异序列构建转化子,以c-erbB-2的特异序列为探针进行间期核FISH检测乳腺肿瘤组织中c-erbB-2基因的扩增,并采用基因组半定量法比较不同组别乳腺组织中c-erbB-2基因的扩增。结果FISH检测发现正常乳腺组织中未出现c-erbB-2扩增,而良性与恶性组织中有不同程度的扩增,基因组半定量分析显示恶性肿瘤组织中c-erbB-2扩增程度高于良性与正常组织。结论c-erbB-2扩增是乳腺癌变的可靠的分子指标。可用荧光原位杂交与基因组半定量法对乳腺癌进行分子生物学诊断,且前者的敏感性与稳定性优于后者。     

Development of humanized bispecific antibodies reactive with cytotoxic lymphocytes and tumor cells overexpressing the HER2 protooncogene

The HER2 protooncogene encodes a 185-kD transmembrane phosphoglycoproteins, human epidermal growth factor receptor 2 (p185HER2), whose amplified expression on the cell surface can lead to malignant transformation. Overexpression of HER2/p185HER2 is strongly correlated with progression of human ovarian and breast carcinomas. Recent studies have shown that human T cells can be targeted with bispecific antibody to react against human tumor cells in vitro. We have developed a bispecific F(ab')2 antibody molecule consisting of a humanized arm with a specificity to p185HER2 linked to another arm derived from a murine anti-CD3 monoclonal antibody that we have cloned from UCHT1 hybridoma. The antigen-binding loops for the anti-CD3 were installed in the context of human variable region framework residues, thus forming a fully humanized BsF(ab')2 fragment. Additional variants were produced by replacement of amino acid residues located in light chain complementarity determining region 2 and heavy chain framework region 3 of the humanized anti-CD3 arm. Flow cytometry analysis showed that the bispecific F(ab')2 molecules can bind specifically to cells overexpressing p185HER2 and to normal human peripheral blood mononuclear cells bearing the CD3 surface marker. In additional experiments, the presence of bispecific F(ab')2 caused up to fourfold enhancement in the cytotoxic activities of human T cells against tumor cells overexpressing p185HER2 as determined by a 51Cr release assay. These bispecific molecules have a potential use as therapeutic agents for the treatment of cancer.     

Detection of the extracellular domain of c-erbB-2 oncoprotein in sera from patients with various carcinomas: Correlation with tumor markers

Using a serum enzyme immunoassay (EIA) kit from Triton diagnostics we detected c-erbB-2 oncoprotein activity in random sera containing highly elevated tumor markers and also in serial specimens from cancer patients expressing elevated oncoprotein activities. Elevated oncoprotein activity was found not only in sera of breast and ovarian carcinomas but also in sera from colorectal, pancreatic, and prostate carcinomas and even from primary hepatoma. Whenever oncoprotein was overexpressed in an individual patient, there was usually an excellent correlation between the oncoprotein activity and the level of dominant tumor marker in serial serum specimens. Based on the size exclusion S-200 column chromatography, we found only a single molecule containing c-erbB-2 oncoprotein activity in pooled sera from cancer patients whereas two oncoproteins slightly different in size were detected in breast tumor tissue cytosol. Using HPLC on a Superose 12 HR column, the serum portion of the oncoprotein was eluted at a position near IgG, suggesting that the extracellular domain of the oncoprotein exists as a dimer in the serum. © 1993 Wiley-Liss, Inc.     

Ⅰ型生长因子受体家族在非小细胞肺癌中的表达及其预后价值的研究

目的 探讨Ⅰ型生长因子受体家族在非小细胞肺癌（NSCLC）组织中异常表达的临床意义及预后价值。方法 用免疫组化法研究了58例NSCLC组织中c-erbB-1、c-erbB-2、c-erbFk3、c—erbB-4蛋白的表达。结果 NSCLC组织中,C-erbB-1、C-erbD-2、c—erbB-3蛋白表达高于正常组织,过表达率分别为62．07％、60．34％和41．38％。C-erbB-4表达率为63．85％,显著低于正常组织（P=0．03）。c—erbB-1蛋白过表达与PCNA指数呈正相关（r=0．248,P〈0．05）;c-erbB-2蛋白过表达与肿瘤的分化程度呈负相关（r=0．29,P〈0．05）,c-erbB-2和C-erbB-3蛋白的过表达,及二者的共过表达,与TNM分期有密切关系（P〈0．01）;C-erbB-4蛋白表迭趋向于高分化肿瘤（P〈0．01）。C-erbB-2蛋白过表达及其与c—erbB-3蛋白共过表达显示预后不良（P〈0．01）。结论 Ⅰ型生长因子受体家族在NSCLC的发生、发展中起着重要的作用,并有助于判断预后。     

Identification of mutations in the putative ATP-binding domain of the adrenoleukodystrophy gene.

We evaluated the prognostic significance of p185c-erbB-2 expression and ras gene mutations in all patients diagnosed with a pulmonary adenocarcinoma between 1982 and 1985 at the University of Iowa. p185c-erbB-2 expression was detected in 15 cases (34%). A ras gene mutation was found in 16 cases (36%) and all were in codon-12 of K-ras. No N-ras mutations were identified. Both p185c-erbB-2 expression and a K-ras mutation were found only in codon-12 and present in six cases (14%). By univariate analysis p185c-erbB-2 expression was associated with shortened survival (P = 0.02) while the presence of a K-ras mutation was not (P = 0.16). Multivariate analysis by the Cox proportional hazards model, controlling for patient age and tumor stage, also continued to identify p185c-erbB-2 expression as an independent unfavorable prognostic factor (P = 0.01). In this model a K-ras mutation also approached significance as a poor prognostic indicator (P = 0.06). The impact of both p185c-erbB-2 expression and a K-ras mutation on survival was additive and highly significant (P = 0.004). This additive nature suggests that together these two markers identify a high-risk population of lung adenocarcinoma patients that may benefit from aggressive therapy.     

c-erbB-2 and episialin challenge host immune response by HLA class I expression in human non-small-cell lung cancer

The role of major histocompatibility complex expression in cancer prognosis and pathogenesis is contradictory. The aim of the current study was to compare the expression of HLA class I molecules and of oncoproteins that may be sources of peptides presented by HLA class I antigens in non-small-cell lung cancer. For this purpose, the expression of HLA class I antigen and TAP-1 molecule (a transporter in the antigen-processing 1 transport protein) were studied with epidermal growth factor, receptor; c-erbB-2; episialin; wild-type and mutant p53; bcl-2 oncoprotein expression; and angiogenic factor expression (vascular endothelial growth factor and thymidine phosphorylase). The degree of lymphocytic stromal infiltration and of platelet-endothelial cell adhesion molecule-expressing lymphocytes was also studied. A strong association of c-erbB-2 and MUC1 (episialin) expression with HLA class I expression was observed (p = 0.005 and 0.009, respectively). Intense CD31-positive lymphocytic infiltration was also more frequent in HLA class I-positive cases (p = 0.05). Although there was no association of HLA class I expression with survival, loss of the HLA class I expression in MUC1 or c-erbB-2 overexpressing cases conferred a poorer clinical outcome (p = 0.04). Both c-erbB-2 and MUC1 are well-known targets of T-cell-mediated cytotoxicity and cell-cell or cell-matrix adhesion-regulating proteins. The authors provide evidence that the sequence of cell adhesion-disrupting oncoprotein expression, HLA class I induction, and enhanced epitope presentation followed by lymphocytic response is an important pathogenetic three-step sequence of events that define, in part, the clinical outcome in non-small-cell lung cancer.     

Psa immunoreactivity detected in lncap cell medium,breast tumor cytosol,and female serum

We made an effort to identify a reliable source for obtaining large quantities of both free (PSA) and PSA–ACT complex for the preparation of the calibrator for the PSA assay. Using size exclusion chromatography, we found both free PSA and PSA–ACT complex in the conditioned cell medium of the LNCaP cell line, which was derived from a human metastatic adenocarcinoma of the prostate. An assay specific for PSA–ACT reacted only with the PSA–ACT complex from cells grown in serum-free medium, and not with the complex from the cell medium grown in 10% calf serum. We also found both free PSA and PSA–ACT complex in 15% of cytosols prepared from breast tumor tissues; the cytosol PSA concentrations ranged from 0.1 to 110 ng/ml. No correlation was found between cytosol PSA and concentrations of estrogen receptor, progestin receptor, epidermal growth factor receptor, cathepsin D, or the ectodomain of c-erbB-2 protein. Based on chromatographic characterizations and the slope of their dose-response curves, it appears that both free PSA and PSA–ACT complex found in the cytosols are similar to PSA complex from the cell medium and the serum of prostate cancer patients. Ectopic PSA was also detected in pooled sera from patients with breast, ovarian, pancreatic, and colon carcinoma. The PSA concentrations in these serum pools increased with the level of their dominant tumor marker. In any event, the LNCaP cell medium appears to be a reliable source for obtaining both free and ACT-complexed PSA of human tumor origin for the preparation of PSA assay calibrators.     

Correlation between polyamines and apoptosis among Egyptian breast cancer patients

OBJECTIVES: The polyamines putrescine, spermidine, and spermine, play an important role in cell proliferation, differentiation, and transformation. The aim of this study is to correlate the polyamines with apoptosis and clinico-pathologic events in Egyptian breast cancer patients. METHODS: PUT, SPD, and SPN were investigated using thin layer chromatography (TLC) and apoptosis in fresh frozen tissue specimens obtained from 40 patients suffering from breast cancer, as well as 20 patients with benign breast lesions. RESULTS: The levels of PUT, SPD, and SPN were higher in breast cancer tissues than in benign breast lesions (p < 0.001). Polyamines were correlated well with apoptosis. Moreover, PUT was an independent prognostic factor for relapse. Also, SPD and SPN correlated significantly with early tumor grades. ROC curves were used to choose the best cut-off values for polyamines (70, 135, and 290 mmol/g tissue) for PUT, SPD, and SPN, respectively. At these cut-off values, the sensitivities were (75%, 60%, and 70%), and the specificities were (80%, 95%, and 95%) for PUT, SPD, and SPN, respectively. CONCLUSION: Polyamines may be used as additional markers for detection of malignant transformation in breast tissue. Moreover, because of their ability to induce apoptosis in malignant tissues, polyamines are suitable targets for therapeutic intervention that is specifically directed to induce apoptosis.     

Development of two ELISA for estrogen and progesterone receptor with sufficient sensitivity for fine needle aspirate and core biopsy

The quantitative determination of estrogen and progesterone receptors (PR and ER) in breast tumor cytosol has been routinely performed in clinical laboratories to aid in the selection between hormonal and chemotherapy and also to predict prognosis. However, the small amount of tissue available from the increasingly popular fine-needle aspiration and core biopsies from breast cancer patients requires more sensitive immunoassays for receptor quantification. We have developed two sensitive immuno-assays for ER and PR on microplate with the use of recently available anti-ER and anti-PR antibodies of higher affinity and a powerful signal magnification agent, namely Amdex. The calibrator was a pooled breast tumor cytosol used as calibrator and calibrated against Abbott kits. The protein concentration of the cytosol and the upper normal cutoffs for our assays were reduced to approximately 0.2 mg/mL and 3 fmol/0.2 mg/mL, respectively. Both assays have sensitivities close to 1 fmol/mL, which are sufficiently sensitive for the receptor quantification in fine-needle aspiration biopsies and cord biopsies of breast tumor.     

HER2 testing: a review of detection methodologies and their clinical performance

The ERBB2 proto-oncogene, commonly referred to as the human epidermal growth factor receptor-2 (HER2) gene, encodes a 185 kd receptor tyrosine kinase. Overexpression of the protein leads to constitutive activity of the HER2 receptor and breast tumor development through enhanced cell proliferation, survival, motility and adhesion. Overabundance of the HER2 receptor, typically caused by amplification of the HER2 gene, is present in approximately 10-30% of invasive breast cancers, and is associated with an aggressive disease course and decreased disease-free and overall survival in node-positive patients. Tratuzumab, a humanized murine monoclonal antibody, offers a targeted treatment modality for tumors that over express the HER2 protein. Tratuzumab, shown to be effective and initially approved for treatment of metastatic breast cancer, has recently been shown to be very effective in the adjuvant setting. Thus, to offer prognostic information and to direct appropriate treatment it is important to provide accurate laboratory assessment of the status of HER2. This article provides an overview of the methods currently used to assess HER2.     

乳腺浸润性导管癌c-erbB-2的表达与淋巴结转移和肿瘤细胞分裂能力的关系

目的探讨乳腺浸润性导管癌中c-erbB-2与肿瘤生物学特性的关系。方法应用免疫组化SP方法对检测乳腺浸润性导管癌c-erbB-2的表达情况极其淋巴结转移和癌细胞分裂能力的相关性。结果c-erbB-2阳性表达主要分布在肿瘤细胞的胞膜上;随着c-erbB-2表达强度的降低,肿瘤淋巴结转移率呈减少的趋势;肿瘤组织中细胞的核分裂数目随着c-erbB-2的表达强度增加而增多。结论c-erbB-2是乳腺癌细胞增殖、转移能力和预后判定的参考指标。