Study of c-erbB-2 protein and epidermal growth factor receptor expression and DNA ploidy pattern in colorectal carcinoma

Correlation of c-erbB-2 protein (n = 44), epidermal growth factor receptor (EGFR) (n = 41) expression, and DNA ploidy pattern (n = 42) with clinical outcomes of human colorectal cancers was studied. Using monoclonal antibodies against c-erbB-2 protein and EGFR, an immunohis-tochemical study of the expression of c-erbB-2 protein and EGFR in frozen tissue sections from the lesion was performed. There was no significant correlation between the expression of c-erbB-2 protein and clinicopatho-logical findings such as, tumor size, histological type, depth of invasion, lymph node metastasis, lymphatic vessel invasion, or venous invasion. However, the incidence of c-erbB-2 protein expression in Dukes D was significantly higher (9/10, 90%) than that in Dukes A to C (16/34, 47.1%). Similar tendency was also observed in the expression of EGFR. Aneuploid case was observed in 12 of observed 25 (48%) cases without lymph node metastasis, while it was observed in 16 of 17 cases (94.1%) with lymph node metastasis and there was significant association between DNA ploidy pattern and lymph node metastasis (P< 0.01) and most of the cases (17/20, 85%) were aneuploidy in Dukes C and D. The results above suggest that the expression of c-erbB-2 protein or EGFR was associated with distant metastasis, while on the other hand DNA ploidy pattern was correlated with lymph node metastasis. © 1993 Wiley-Liss, Inc.     

Expression of c-erbb-2 protein and epidermal growth factor receptor in endometrial carcinomas correlation with clinicopathologic and sex steroid receptor status

Background. The c-erbB-2 (HER-2/neu) protein is a membrane glycoprotein growth factor receptor showing molecular homology with the epidermal growth factor receptor (EGFR). In endometrial carcinomas, little is known about the relationship between the expression of c-erbB-2 protein and that of EGFR. Methods. The immunohistochemical reactivity of monoclonal antibodies against both of these proteins was examined in 34 endometrial carcinomas, and the presence or absence of correlation with the clinicopathologic features or with the immunohistochemical expression of sex steroid receptors (estrogen receptor [ER] and progesterone receptor [PR]) was analyzed. Results. Of the 34 patients, 22(64.7%) had c-erbB-2 protein-positive and EGFR-negative tumor, and 8 (23.5%) had tumor positivity for both proteins. Four patients had tumors negative for both proteins. ER or PR positivity was found in 24 (70.6%) of the 34 patients. Intense immunostaining for c-erbB-2 protein was found in 5 (14.7%) of the 34 patients but was not correlated with the stage or grade of differentiation in endometrial carcinoma. However, expression of EGFR in addition to c-erbB-2 protein was more frequently observed with advancing stage of disease and was inversely correlated with the grade of differentiation and with the expression of ER or PR of the tumor. Conclusion. The expression of EGFR, in addition to that of c-erbB-2 protein, is an important event that presumably is linked with progression or with a poorly differentiated state of endometrial carcinomas.     

Significant intratumoral heterogeneity of human epidermal growth factor receptor 2 status in gastric cancer: A comparative study of immunohistochemistry,FISH, and dual‐color in situ hybridization

The assessment of human epidermal growth factor receptor 2 (HER2) status is crucial for selecting patients with gastric cancer who may benefit from HER2‐targeted therapy. Accurate assessment using biopsy specimens is important for patients with advanced‐stage cancer. Intratumoral heterogeneity of HER2, however, is a major challenge in HER2 testing. Here, we aimed to examine whether assessment of HER2 status could be accurately carried out with currently used methods, namely, immunohistochemistry (IHC), FISH, and dual‐color in situ hybridization (DISH). Human epidermal growth factor receptor 2 status was evaluated in 108 biopsy tissues from patients with gastric adenocarcinoma and 70 matched surgical specimens by IHC, FISH, and DISH; HER2 amplification was detected in 11 (10.2%) out of 108 biopsy specimens. The IHC and FISH results were well correlated, and FISH and DISH results were consistent for all cases. The overall concordance rate of HER2 status between biopsy tissues and surgical specimens was 91.4%. All six discordant cases were false negative on biopsy; of these cases, five showed HER2 heterogeneity on surgical resection. Assessment of the HER2 status of biopsy tissues could predict the status of the whole tumor; however, a proportion of these cases may be discordant because of intratumoral heterogeneity.     

Comparative analysis of epidermal growth factor receptor mutations and gene amplification as predictors of gefitinib efficacy in Japanese patients with nonsmall cell lung cancer

BACKGROUND: Because the investigation of epidermal growth factor receptor gene (EGFR) status as a predictor of gefitinib efficacy in Japanese patients has shown promise, the authors evaluated EGFR mutations and gene amplification in biopsy specimens from Japanese patients with nonsmall cell lung cancer (NSCLC) who received treatment with gefitinib to analyze the correlation between EGFR gene status and clinical outcome. METHODS: Fifty-nine patients were enrolled in this study. EGFR gene amplification was evaluated by fluorescence in situ hybridization (FISH), and EGFR mutations in exons 18, 19, and 21 were analyzed by polymerase chain reaction and direct sequencing. RESULTS: EGFR mutations were detected in 17 patients (28.8%). FISH-positive results were observed in 26 patients (48.1%). The response rate was significantly higher in the patients with EGFR mutations than in the patients without mutations (58.8% vs 14.3%; P=.0005). No significant difference in the response rate was observed between FISH-positive patients and FISH-negative patients (31.8% vs 21.4%; P=.4339). EGFR mutation was correlated with both a longer time to progression (TTP) (7.3 months vs 1.8 months; P=.0030) and longer overall survival (OS) (18.9 months vs 6.4 months; P=.0092). No significant differences in TTP or OS were observed between FISH-positive patients andFISH-negative patients. The results from a multivariate analysis indicated that EGFR mutations maintained a significant association with longer TTP and longer OS. CONCLUSIONS: The results of this study suggested that EGFR mutations may serve as predictors of response and survival and that the role of EGFR gene amplification is not a predictor of gefitinib efficacy in Japanese patients with NSCLC.     

Breast cancers presenting luminal B subtype features show higher discordant human epidermal growth factor receptor 2 results between immunohistochemistry and fluorescence in situ hybridization

BACKGROUND:

The aims of this study were to compare human epidermal growth factor receptor 2 (HER2) results between immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), and to investigate the clinicopathological characteristics and outcomes according to their results.

METHODS:

Using consecutive tissue microarrays, IHC and FISH were performed as guidelines in 950 invasive breast cancers treated between November 1999 and August 2005. Characteristics and outcomes were retrospectively analyzed using a chi‐square test, the Kaplan‐Meier method, and Cox's model.

RESULTS:

FISH‐positivity was observed in 2.6%, 4.8%, 28.1%, and 93.8% of IHC 0, 1+, 2+, and 3+, respectively, and the concordance rate between the 2 assays was 95.5%. IHC‐positive or FISH‐positive cases were associated with poorer differentiation, negative expression of hormone receptors, and higher proliferative index. Among IHC‐equivocal or IHC‐negative patients, positive FISH was negatively associated with survival in univariate and multivariate analyses. Among IHC‐negative patients, tumors showing luminal B subtype features such as estrogen receptor (ER)‐positive, grade II/III, and high Ki‐67 presented discordantly high FISH‐positivity. Among IHC‐positive cases, FISH was not related to outcomes.

CONCLUSIONS:

The result of FISH is significantly related to prognosis of patients with IHC‐negative or IHC‐equivocal result. Therefore, FISH should be performed in IHC‐equivocal cases. FISH assay might be considered for a selected group of patients with IHC‐negative tumors showing luminal B subtype features of ER‐positive, grade II/III, and high Ki‐67 expression. Cancer 2012;. © 2011 American Cancer Society.     

Expression of epidermal growth factor receptor mRNA and protein in primary breast carcinomas

The expression of epidermal growth factor receptor (EGFR) mRNA and protein has been determined in a group of breast carcinomas and compared to oestrogen and progesterone receptor (ER, PgR) status, as well as pathological features. In situ hybridization using a digoxigenin-labelled oligonucleotide probe was applied to formalin-fixed paraffin-embedded sections, and immunohistochemistry was used to determine EGFR protein.EGFR mRNA was detected in 66% of carcinomas with a third having labelling similar to normal breast tissue, 22% heterogeneous weak to strong labelling, and 11% strong labelling. EGFR protein was detected in 36% and these tumours had a strong correlation to lack of ER and high histological grade. The presence of EGFR protein was strongly correlated with more intense labelling for EGFR mRNA (p < 0.0001). This contrasted with normal breast in which both EGFR protein and mRNA were present with varying degrees in both tumours and a normal breast control. The ER-/PgR- carcinomas showed the full range of EGFR mRNA labelling. It is postulated that oestrogen or oestrogen regulated proteins are involved in regulation of EGFR mRNA and protein. In a proportion of tumours lacking steroid receptors regulation is lost, leading to EGFR overexpression.     

胃腺癌内镜活检和术后标本HER-2 表达的一致性分析

目的:分析胃腺癌内镜活检和术后标本HER-2 蛋白表达状态的一致性,探讨胃腺癌内镜活检标本HER-2 检测结果对应用曲妥珠单抗治疗的指导价值。方法:选取2013年3 月至2014年2 月上海长海医院病理科诊断明确的胃腺癌内镜活检标本及相应的肿瘤根治手术切除标本167 例,并收集相关临床病理资料。采用免疫组织化学（immuno histochemistry,IHC ）方法检测内镜活检标本的HER-2 蛋白表达情况,用IHC 及原位荧光杂交（fluorescence in situ hybridization,FISH）方法检测相应肿瘤手术切除标本的HER-2 蛋白表达和基因扩增状况,对检测结果进行比较,并结合临床病理特征进行分析。结果:167 例根治标本中,共有18例（10.8%）HER-2 检测呈阳性,其中包含10例IHC 3 + 与8 例IHC 2 + 且FISH+ 的病例。相对应的内镜活检标本与根治手术标本的IHC 检测结果的一致率为82.0% ,剔除IHC 2 + 的样本后,真阳性率和真阴性率分别为73.3% 和97.0% 。结论:内镜活检标本HER-2蛋白IHC 检测具有较好预测价值,根治与活检标本不一致的主要原因是肿瘤表达的异质性。通过提高内镜活检标本取材数,并结合FISH检测结果,可以减少误判。      

The type 1 growth factor receptor family: new ligands and receptors and their role in breast cancer

The type 1 family of growth factor receptors, which consist of the epidermal growth factor receptor, c-erbB-2, c-erbB-3, and c-erbB-4, are expressed in normal breast ductal epithelial cells and in some breast cancers. Nine genes have now been identified which code for ligands. In some cases the genes are spliced into a series of proteins which differ in structure, but all retain an EGF-like element responsible for receptor recognition.The EGF receptor is expressed in normal breast and in some cancers, but is apparently reduced in expression in other cases. Cancers with EGF receptors appear to represent a greater threat to patients as in most studies they are associated with a shorter time to relapse and overall survival. The c-erbB-2 protein is overexpressed at very high levels in about one fifth of breast cancers and is indicative of poor prognosis. Other cancers may express lesser degrees of overexpression but it is not clear if this is biologically or clinically significant. The c-erbB-3 protein is expressed in normal breast epithelial cells and has been reported to be present at high levels in some cancers but at normal levels or at lower than normal levels in some others. The limited studies to date suggest that when measured on its own c-erbB-3 expression is not predictive. c-erbB-4 is also expressed in normal breast and in some cancers but no studies have yet been performed to address whether it is associated with disease behaviour.In the future it is likely that a greater understanding of the function of this complex family of interacting proteins will assist in gaining the maximum predictive power from measurement of their expression in human breast cancer.     

人表皮生长因子受体-2及雌激素受体与乳腺癌的关系

乳腺癌与人表皮生长因子受体-2及雌激素受体关系密切,这两种受体也是乳腺癌的分类标准和治疗靶点。在大多数乳腺癌患者中,人表皮生长因子受体-2信号途径和雌激素受体信号途径参与了细胞的增生存活过程。而且在乳腺癌病例中,人表皮生长因子受体-2和雌激素受体呈现出一定程度的负相关。这说明这两种受体活化后有一些联系。本文简要综述了人表皮生长因子受体-2和雌激素受体的联系以及这种联系在乳腺癌治疗中的意义。     

Role of epidermal growth factor receptor expression in primary breast cancer: results of a biochemical study and an immunocytochemical study

Summary We have conducted two series of studies, a biochemical study and an immunocytochemical study, to investigate the role of epidermal growth factor receptor (EGFR) expression in primary breast cancer patients. In the biochemical study, a consecutive 115 patients were included and EGFR was measured by a competitive binding assay with multipoint Scatchard analysis. In the immunocytochemical study comprising 126 patients, EGFR status was determined by immunostaining with anti-EGFR antibody EGFR1. Several agreements were found from these two studies. EGFR status was inversely correlated with estrogen receptor (ER) status. No significant correlation was found between EGFR status and tumor size, nodal metastases, or the expression of c-erbB-2 protein. Ki-67 immunoreactivity, a cellular proliferation marker, was enhanced in EGFR positive tumors over EGFR negative tumors, suggesting a linkage of EGFR expression to cellular proliferative activity. Post-operative follow up showed that relapse-free survival for EGFR positive patients was significantly worse than that for EGFR negative patients, particularly in node-positive patients. Multivariate analysis demonstrated a significance of EGFR status as an independent prognostic indicator in primary breast cancer. The group expressing EGFR and c-erbB-2 protein indicated a particularly high risk for relapse.     

Impact of chromosome 17 centromere copy number increase on patient survival and human epidermal growth factor receptor 2 expression in gastric adenocarcinoma

The accurate evaluation of human epidermal growth factor receptor 2 (HER2) status is essential for the appropriate use of targeted therapies. An increased number of chromosome 17 centromere enumeration probe (CEP17) signals may underrate fluorescence in situ hybridization (FISH) outcomes, resulting in false-negative or a false-equivocal HER2 status assessment. The aim of the present study was to assess the frequency of CEP17 copy number increase (CNI), its effects on HER2 protein expression (and the subsequent effects on tumor cells), and the survival outcomes of patients with gastric cancer. Archival primary tumor samples from 244 patients that underwent gastric resection for adenocarcinoma were retrieved for both HER2 protein expression analysis (using immunochemistry) and HER2 gene amplification (using FISH). The associations between HER2 status, CEP17 CNI and multiple clinicopathological parameters (including survival outcome), were assessed. The relationship between CEP17 CNI and HER2 protein upregulation was also investigated. CEP17 CNI was detected in 17.2% of cases, and a strong association between CEP17 CNI and HER2 upregulation was revealed. The impact of CEP17 CNI on survival did not reach statistical significance. Consequently, CEP17 CNI was discovered to be strongly associated with HER2 upregulation in tumor cells, which may characterize a critical issue in HER2 testing. Therefore, the eligibility for HER2-targeted agents in CEP17 CNI-positive patients warrants further recognition.     

Cost-effectiveness of epidermal growth factor receptor mutation testing and first-line treatment with gefitinib for patients with advanced adenocarcinoma of the lung

BACKGROUND:

Epidermal growth factor receptor (EGFR) testing and first‐line therapy with gefitinib for patients with activating mutations is quickly becoming the standard option for the treatment of advanced lung adenocarcinoma. Yet, to date, little is known about the cost‐effectiveness of this approach.

METHODS:

A decision‐analytic model was developed to determine the cost‐effectiveness of EGFR testing and first‐line treatment with gefitinib for those patients who harbor activating mutations versus standard care, which includes first‐line treatment with chemotherapy followed by gefitinib as second‐line treatment. The model uses clinical and outcomes data from randomized clinical trials and societal costs from Singapore cancer centers. Health effects were expressed as quality‐adjusted life‐years. All costs and cost‐effectiveness ratios were expressed in 2010 Singapore dollars. Sensitivity and different scenarios analyses were conducted.

RESULTS:

EGFR testing and first‐line treatment with gefitinib is a dominant strategy (with lower costs and greater effectiveness) compared with standard care. Because the primary savings result from not providing gefitinib to those who are not likely to benefit, this finding holds regardless of the prevalence of activating mutations. In a secondary analysis, first‐line treatment with gefitinib was also dominant when compared with first‐line chemotherapy in patients with activating EGFR mutations.

CONCLUSIONS:

This strategy can be considered a new standard of care and should be of great interest for health care payers and decision makers in an era in which our greatest challenge is to balance hard‐won and incremental, yet small, improvements in patient outcomes with exponentially rising costs. Cancer 2012;. © 2011 American Cancer Society.     

表皮生长因子、雄激素受体和表皮生长因子受体在肝癌组织中的表达及影响因素分析

目的探讨肝癌患者肝癌组织中的表皮生长因子(EGF)、雄激素受体(AR)、表皮生长因子受体(EGFR)的表达情况及临床意义。方法采用免疫组织化学染色法检测EGF、AR、EGFR在90例肝癌患者的肝癌组织和90例非肝癌患者的非肝癌组织中的表达情况,比较不同临床特征肝癌患者肝癌组织中EGF、AR、EGFR的表达情况,分析肝癌患者肝癌组织中EGF、AR、EGFR表达的影响因素。结果肝癌组织中的EGF、AR、EGFR的阳性表达率均明显高于非肝癌组织,差异均有统计学意义(P﹤0.01)。有肝炎史、Ⅲ+Ⅳ期、中低分化、有淋巴结转移的肝癌患者肝癌组织中EGF、AR、EGFR的阳性表达率均高于无肝炎史、Ⅰ+Ⅱ期、高分化、无淋巴结转移的肝癌患者,差异均有统计学意义(P﹤0.05);不同年龄、性别、肿瘤直径的肝癌患者肝癌组织中EGF、AR、EGFR的阳性表达率比较,差异均无统计学意义(P﹥0.05)。Logistic回归分析结果显示,有淋巴结转移、TNM分期为Ⅲ+Ⅳ期是肝癌患者肝癌组织中EGF、AR、EGFR表达的独立危险因素。结论EGF、AR、EGFR在肝癌患者的肝癌组织中呈高表达,且其表达与肝癌患者的淋巴结转移情况和TNM分期密切相关。     

Systemic therapy for hormone receptor-positive/human epidermal growth factor receptor 2-negative early stage and metastatic breast cancer

Hormone receptor (HR)-positive and human epidermal growth factor receptor 2 (HER2)-negative breast cancer is defined by the presence of the estrogen receptor and/or the progesterone receptor and the absence of HER2 gene amplification. HR-positive/HER2-negative breast cancer accounts for 65%–70% of all breast cancers, and incidence increases with increasing age. Treatment varies by stage, and endocrine therapy is the mainstay of treatment in both early stage and late-stage disease. Combinations with cyclin-dependent kinase 4/6 inhibitors have reduced distant recurrence in the early stage setting and improved overall survival in the metastatic setting. Chemotherapy is used based on stage and tumor biology in the early stage setting and after endocrine resistance for advanced disease. New therapies, including novel endocrine agents and antibody-drug conjugates, are now changing the treatment landscape. With the availability of new treatment options, it is important to define the optimal sequence of treatment to maximize clinical benefit while minimizing toxicity. In this review, the authors first discuss the pathologic and molecular features of HR-positive/HER2-negative breast cancer and mechanisms of endocrine resistance. Then, they discuss current and emerging therapies for both early stage and metastatic HR-positive/HER2-negative breast cancer, including treatment algorithms based on current data.     

Preclinical antitumor activity of S‐222611, an oral reversible tyrosine kinase inhibitor of epidermal growth factor receptor and human epidermal growth factor receptor 2

Epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) are validated molecular targets in cancer therapy. Dual blockade has been explored and one such agent, lapatinib, is in clinical practice but with modest activity. Through chemical screening, we discovered a novel EGFR and HER2 inhibitor, S‐222611, that selectively inhibited both kinases with IC₅₀s below 10 nmol/L. S‐222611 also inhibited intracellular kinase activity and the growth of EGFR‐expressing and HER2‐expressing cancer cells. In addition, S‐222611 showed potent antitumor activity over lapatinib in a variety of xenograft models. In evaluations with two patient‐oriented models, the intrafemoral implantation model and the intracranial implantation model, S‐222611 exhibited excellent activity and could be effective against bone and brain metastasis. Compared to neratinib and afatinib, irreversible EGFR/HER2 inhibitors, S‐222611 showed equivalent or slightly weaker antitumor activity but a safer profile. These results indicated that S‐222611 is a potent EGFR and HER2 inhibitor with substantially better antitumor activity than lapatinib at clinically relevant doses. Considering the safer profile than for irreversible inhibitors, S‐222611 could be an important option in future cancer therapy.     

人表皮生长因子受体2模拟表位的筛选

目的:从噬菌体12肽库中筛选出人表皮生长因子受体2（Her2）的抗原模拟表位。方法:以曲妥珠单抗为靶分子,在噬菌体12肽库中进行3轮淘选,以ELISA方法及竞争抑制实验鉴定阳性克隆,并对阳性克隆株进行测序。结果:经过3轮淘选,与曲妥珠单抗结合的噬菌体得到了有效富集,回收率从（2.00×10-8）%增加到（2.87×10-5）%,ELISA显示20个克隆中筛选获得了18个与曲妥珠单抗具有较高亲和性的阳性噬菌体,对阳性克隆测序获得两种氨基酸序列:HTSSLWHLFRST、VHWDFRQWWQPS。结论:噬菌体展示技术可成功筛选到表皮生长因子2模拟表位,为探索乳腺癌的防治研究创造了条件。