Pre-existent immunity to the HER-2/neu oncogenic protein in patients with HER-2/neu overexpressing breast and ovarian cancer

Immunomodulatory strategies, such as antibody therapy and cancer vaccines, are increasingly being considered as potential adjuvant therapies in patients with advanced stage breast cancer to either treat minimal residual disease or prevent relapse. However, little is known concerning the incidence and magnitude of the pre-existent breast cancer specific immune response in this patient population. Using the HER-2/neu oncogenic protein as a model, a well-defined tumor antigen in breast cancer, we questioned whether patients with advanced stage HER-2/neu overexpressing breast and ovarian cancers (III/IV) had evidence of pre-existent immunity to HER-2/neu. Forty-five patients with stage III or IV HER-2/neu overexpressing breast or ovarian cancer were evaluated for HER-2/neu specific T cell and antibody immunity. Patients enrolled had not received immunosuppressive chemotherapy for at least 30 days (median 5 months, range 1–75 months). All patients were documented to be immune competent prior to entry by DTH testing using a skin test anergy battery. Five of 45 patients (11%) were found to have a significant HER-2/neu specific T cell response as defined by a stimulation index 2.0 (range 2.0–7.9). None of eight patients who were HLA-A2 had a detectable IFN secreting T-cell precursor frequency to a well-defined HER-2/neu HLA-A2 T cell epitope, p369-377. Three of 45 patients (7%) had detectable HER-2/neu specific IgG antibodies, range 1.2–8.9g/ml. These findings suggest that patients with advanced stage HER-2/neu overexpressing breast and ovarian cancer can mount a T cell and/or antibody immune response to their tumor. However, in the case of the HER-2/neu antigen, the pre-existent tumor specific immune response is found only in a minority of patients.     

Modest effect of p53, EGFR and HER-2/neu on prognosis in epithelial ovarian cancer: a meta-analysis

Background:

P53, EGFR and HER-2/neu are the most frequently studied molecular biological parameters in epithelial ovarian cancer, but their prognostic impact is still unequivocal. We performed a meta-analysis to more precisely estimate their prognostic significance.

Methods:

Published studies that investigated the association between p53, EGFR and HER-2/neu status and survival were identified. Meta-analysis was performed using a DerSimonian–Laird model. Publication bias was investigated using funnel plots and sources of heterogeneity were identified using meta-regression analysis.

Results:

A total of 62 studies were included for p53, 15 for EGFR and 20 for HER-2/neu. P53, EGFR and HER-2/neu status had a modest effect on overall survival (pooled HR 1.47, 95% CI 1.33–1.61 for p53; HR 1.65, 95% CI 1.25–2.19 for EGFR and HR 1.67, 95% CI 1.34–2.08 for HER-2/neu). Meta-regression analysis for p53 showed that FIGO stage distribution influenced study outcome. For EGFR and HER-2/neu, considerable publication bias was present.

Conclusions:

Although p53, EGFR and HER-2/neu status modestly influences survival, these markers are, by themselves, unlikely to be useful as prognostic markers in clinical practice. Our study highlights the need for well-defined, prospective clinical trials and more complete reporting of results of prognostic factor studies.     

卵巢癌HER-2／neu基因表达的临床意义

目的:评价卵巢癌HER-2/neu表达的临床意义及对近期化疗效果的影响。方法:免疫组织化学ABC法检测49例卵巢癌HER-2/neu的表达。结果:1.卵巢癌HER-2/neu表达的阳性率为75.5％,表达阳性者的近期化疗有效率为24.3％,明显低于表达阴性者的近期化疗有效率75.0％(P<0.005)。2.HER-2/neu的表达与卵巢癌分化程度和临床分期密切相关(P<0.01,P<0.005)。3.HER-2/neu过表达的肿瘤具有远位转移的更大风险。结论:卵巢癌HER-2/neu的过表达明显影响近期化疗效果,并可能是其预后不良的肿瘤标志。     

Long-term prognostic significance of HER-2/neu in untreated node-negative breast cancer depends on the method of testing

IntroductionThe prognostic significance of HER-2/neu in breast cancer is a matter of controversy. We have performed a study in 101 node-negative breast cancer patients with long-term follow-up not treated in the adjuvant setting, and analysed the prognostic significance of immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH), both separately and in combination, in comparison with traditional prognostic factors.MethodsOverexpression was classified semiquantitatively according to a score (0 to 3+) (HER-2_SCO). FISH was used to analyse HER2/neu amplification (HER-2_AMP). Patients classified 2+ by IHC were examined with FISH for amplification (HER-2_ALG). Patients with 3+ overexpression as well as amplification of HER-2/neu were positive for the combined variable HER2_COM. These variables were compared with tumour size, histological grade and hormone receptor status.ResultsHER-2_SCO was 3+ in 20% of all tumours. HER-2_ALG was positive in 22% and amplification (HER-2_AMP) was found in 17% of all tumours. Eleven percent of the tumours showed simultaneous 3+ overexpression and amplification. Only histological grade (relative risk [RR] 3.22, 95% confidence interval [CI] 1.73–5.99, P = 0.0002) and HER-2_AMP (RR 2.47, 95% CI 1.12–5.48, P = 0.026) were significant for disease-free survival in multivariate analysis. For overall survival, both histological grade (RR 3.89, 95% CI 1.77–8.55, P = 0.0007) and HER-2_AMP (RR 3.08, 95% CI 1.24–7.66, P = 0.016) retained their independent significance.ConclusionThe prognostic significance of HER-2/neu in node-negative breast cancer depends on the method of testing: only the amplification of HER-2/neu is an independent prognostic factor for the long-term prognosis of untreated node-negative breast cancer.     

乳腺癌中COX-2和HER-2/neu共表达现象及其临床意义

目的观察乳腺癌患者中COX-2的表达及其与HER-2/neu表达的关系,分析COX-2表达对乳腺癌患者DFS的影响。方法应用免疫组化方法检测179例原发性乳腺癌患者标本的COX-2表达及HER-2/neu表达。结果共有25例(13.9%)标本中COX-2呈过表达,并且与HER-2/ neu表达(61/179,34.1%)显著相关(P=0.013)。在单因素生存分析中,COX-2过表达与无疾病生存期显著相关(P=0.029),而COX-2与HER-2/neu共表达患者也有无疾病生存期缩短的趋势。但是多因素分析提示,COX-2过表达与无疾病生存期无显著关系(RR 0.907,95%CI 0.309～2.65,P= 0.858)。结论在人类乳腺癌中,COX-2与HER-2/neu存在共表达。而COX-2与HER-2/neu共表达提示预后差。因此,COX-2选择性抑制剂的临床应用可能是乳腺癌患者治疗的一种新策略。     

PTEN和HER-2／neu在上皮性卵巢癌组织中的表达及相关性

目的探讨卵巢癌组织中PTEN和HER-2／neu的表达与肿瘤行为的关系及意义。方法采用免疫组织化学S-P法检测了158例卵巢癌，20例卵巢良性肿瘤和20例正常卵巢组织中PTEN蛋白和HER-2／neu的表达。结果卵巢癌组织PTEN阳性率（34．18％）明显低于正常卵巢组织（90．00％）和卵巢良性肿瘤组织（80．00％）。卵巢癌组织HER-2／neu过表达率显著高于卵巢正常组织和卵巢良性肿瘤（P〈0．01）。卵巢癌组织中PTEN阳性表达强度与HER-2／neu过表达强度存在显著的负相关。PTEN、HER-2／neu阳性率与卵巢癌的组织分化程度、病理分期有关。结论PTEN蛋白表达的缺失和HER-2／neu过表达可能导致细胞增殖失控，与卵巢癌的发生、发展有关。     

HER-2/neu肿瘤疫苗的研究进展

摘 要　原癌基因HER-2/neu在多种恶性肿瘤中有扩增及过表达,近30%的原发性乳腺癌患者有HER-2/neu过表达。针对HER-2靶点的特异性人源化抗体曲妥株单抗（trastuzumab;又名赫赛汀,Herceptin）在上世纪90年代已用于临床治疗。目前针对HER-2的肽疫苗、蛋白疫苗、细胞疫苗、DC相关疫苗、以及DNA疫苗等的研究,均已取得一定进展,如小肽E75（p369-399）与GM-CSF联合应用,在HLA-A2(+)/A3(+)乳腺癌患者的Ⅱ期临床试验中显示出一定疗效;针对编码HER-2的DNA疫苗已经进入临床试验阶段,这些疫苗均有可能成为乳腺癌治疗的又一个重要手段。但这些疫苗距临床实际使用尚有一定的距离,有许多问题有待解决和需要进一步的临床验证。     

Serological and immunohistochemical HER‐2/neu statuses do not correlate and lack prognostic value for ovarian cancer patients

HOOPMANN M., SACHSE K., VALTER M.M., BECKER M., NEUMANN R., ORTMANN M., GÖHRING U.‐J., THOMAS A., MALLMANN P. & SCHÖNDORF T. (2010) European Journal of Cancer Care
Serological and immunohistochemical HER‐2/neu statuses do not correlate and lack prognostic value for ovarian cancer patients The serodiagnostics of extracellular domain (ECD) HER‐2/neu has turned into an evidenced‐based tumour marker for HER‐2/neu‐positive breast cancer patients. This study investigated the clinical relevance of immunohistochemical and serum HER‐2/neu in 44 patients with advanced ovarian cancer. The Hercept‐Test® from DAKO Diagnostics was used to analyse immunohistochemical HER‐2/neu expression. The HER‐2/neu ECD in serum was determined quantitatively by Bayer Immuno 1? Immunoanalyser. The HER‐2/neu serum values were correlated to the clinical course of disease and to established prognostic factors, i.e. progression‐free and overall survival. Some 23% of patients (n= 11) expressed HER‐2/neu serum levels higher than 15 ng/mL, whereas only 7.7% (n= 2) of the patients examined by immunohistochemistry showed a HER‐2/neu overexpression of the tissue. None of them revealed an overexpression of HER‐2/neu ECD by serodiagnostics. HER‐2/neu overexpression did not correlate significantly to any of the analysed prognostic factors. According to progression‐free and overall survival, there was no significant difference between serologically HER‐2/neu‐positive or negative patients. For ovarian cancer patients, neither high HER‐2/neu serum levels, nor immunohistochemically determined HER‐2/neu positivity, appear to predict the course of disease. This study shows a lack of association between the immunohistochemical HER‐2/neu status and the serum level of solute extracelluar HER‐2/neu domain.     

雌激素受体阳性乳腺癌中孕激素受体表达缺失与HER-2/neu过度表达相关

目的：探究雌激素受体（ER）阳性的乳腺癌中孕激素受体（PgR）表达情况与目前已确立的乳腺癌临床、病理预后指标，包括HER-2／neu表达水平的相关性。方法：对复旦大学附属肿瘤医院2002年6月-2005年6月间连续收治的ER阳性的可手术女性乳腺癌病例资料进行回顾性的分析，获得912例乳腺癌病例作为研究对象。患者年龄24～87岁，中位年龄53岁，其中年龄〈50岁者360例（39．5％），年龄≥50岁552例（60．5％）。采用免疫组化的方法检测ER，PgR和HER-2／neu的表达情况．采用SPSS10．0 for Windows统计软件包进行统计处理。结果：多因素分析显示HER-2／neu过度表达、ER低表达、年龄≥50岁以及淋巴结有转移是独立预测PgR表达缺失的相关因素．本组ER^＋PgR^-乳腺癌较ER^＋PgR^＋乳腺癌存在更多的HER-2／neu过度表达的现象（12．8％ vs 5．8％）；而本组73例（8．0％）HER-2／neu过度表达（“＋＋＋”）的乳腺癌中有50．7％同时合并PgR^-，而HER-2／neu无过度表达（“-～＋＋”）的病例合并PgR的占29．9％，P〈0．001。结论：PgR表达缺失与HER-2／neu过度表达呈显著相关性。     

ＨＥＲ-２／ｎｅｕ与胃癌临床病理及预后的相关性研究

目的　研究ＨＥＲ-２／ｎｅｕ基因蛋白表达与胃癌临床病理特征以及预后的相关性,探讨该基因在胃癌中作为预后因素的价值。方法　应用免疫组织化学染色法对２２９例胃癌组织芯片进行ＨＥＲ-２／ｎｅｕ基因蛋白表达的检测,并将检测结果与患者的临床病理特征以及生存随访结果进行统计分析。结果　２２９例胃癌组织中ＨＥＲ-２／ｎｅｕ基因蛋白表达阳性率为１４.８５％,其表达与患者Ｌａｕｒｅｎ分型、组织学分级、肿瘤大小、淋巴结转移和生存时间相关（Ｐ＜０.０５）;Ｌｏｇ-ｒａｎｋ单因素分析显示,ＨＥＲ-２／ｎｅｕ基因蛋白表达与预后相关（Ｐ＜０.０５）;Ｃｏｘ风险比例模型分析显示,ＨＥＲ-２／ｎｅｕ基因蛋白表达、Ｌａｕｒｅｎ分型、肿瘤大小、脉管浸润和ＴＮＭ分期均为独立预后因素（P＜０.０５）。结论　ＨＥＲ-２／ｎｅｕ基因蛋白表达与胃癌的浸润、转移相关,是胃癌独立的预后因素之一。     

The ErbB signalling pathway: protein expression and prognostic value in epithelial ovarian cancer

Ovarian cancer is the most frequent cause of death from gynaecological cancer in the Western world. Current prognostic factors do not allow reliable prediction of response to chemotherapy and survival for individual ovarian cancer patients. Epidermal growth factor receptor (EGFR) and HER-2/neu are frequently expressed in ovarian cancer but their prognostic value remains unclear. In this study, we investigated the expression and prognostic value of EGFR, EGFR variant III (EGFRvIII), HER-2/neu and important downstream signalling components in a large series of epithelial ovarian cancer patients. Immunohistochemical staining of EGFR, pEGFR, EGFRvIII, Her-2/neu, PTEN (phosphatase and tensin homologue deleted on chromosome 10), total and phosphorylated AKT (pAKT) and phosphorylated ERK (pERK) was performed in 232 primary tumours using the tissue microarray platform and related to clinicopathological characteristics and survival. In addition, EGFRvIII expression was determined in 45 tumours by RT-PCR. Our results show that negative PTEN immunostaining was associated with stage I/II disease (P=0.006), non-serous tumour type (P=0.042) and in multivariate analysis with a longer progression-free survival (P=0.015). Negative PTEN staining also predicted improved progression-free survival in patients with grade III or undifferentiated serous carcinomas (P=0.011). Positive pAKT staining was associated with advanced-stage disease (P=0.006). Other proteins were expressed only at low levels, and were not associated with any clinicopathological parameter or survival. None of the tumours were positive for EGFRvIII. In conclusion, our results indicate that tumours showing negative PTEN staining could represent a subgroup of ovarian carcinomas with a relatively favourable prognosis.     

非小细胞肺癌HER-2/neu基因表达及曲妥珠单抗治疗研究进展

HER-2/neu是一种原癌基因,其高度表达与人肿瘤发生、预后不佳有一定关系。该文就HER-2/neu高度表达与非小细胞肺癌(NSCLC)发生、发展的关系以及单克隆抗体——曲妥珠单抗(Trastuzumab)在临床中对NSCLC的应用价值作一综述。     

Association between HER-2/neu and the progesterone receptor in oestrogen-dependent breast cancer is age-related

Summary In oestrogen receptor-positive (ER⁺) breast cancer, HER-2/neu and the progesterone receptor (PR) are inversely associated. This explains a lower response to anti-oestrogens if ER⁺ breast cancers are HER-2/neu positive. One randomized study however, showed that premenopausal women with an ER⁺ breast cancer respond to anti-oestrogens independent of HER-2/neu. We therefore hypothesized an age-related association between HER-2/neu and PR in ER⁺ breast cancers. Receptors for ER, PR and HER-2/neu were analysed by immunohistochemistry (IHC). A uni- and multivariate analyses was carried out to assess this relationship in 1104 women with an ER⁺ breast cancer. We observed an inverse association between HER-2/neu and PR only after age 45. There were 173 women of age 45 and 931 of age >45 years. In multivariate analysis, only tumour grade (p=0.005) but not PR status was associated with HER-2/neu in women age 45 years. However, in age >45 years group, both PR status (p=0.001) and tumour grade (p = 0.001) were independently associated with HER-2/neu. In ER⁺ breast cancers from women age >45, PR was positive in 76.9% if HER-2/neu negative but in 53.4% if HER-2/neu positive (p<0.001) and the median quantitative PR levels are 150 and 75 respectively in HER-2/neu negative and HER-2/neu positive tumours (p=0.002). This age effect of HER-2/neu on the PR status was not seen in women age 45 years and the median quantitative PR levels are 200 and 220 respectively in HER-2/neu negative and HER-2/neu positive tumours (p = 0.518). The study confirms an age-related inverse relationship between HER-2/neu and PR only in women age >45 years but not in women age 45 years.     

Frequencies of HER-2/neu expression and gene amplification in patients with oesophageal squamous cell carcinoma

The utilisation of antitumour T cells induced by cancer vaccination with HER-2 peptides or antibodies (Herceptin) against HER-2, as immunotherapy for oesophageal cancer, is a novel and attractive approach. It is important to clarify the frequencies of HER-2 expression and gene amplification in patients with oesophageal squamous cell carcinoma (SCC) and to evaluate the relationship between HER-2 status and HLA haplotype, since the candidates for HER-2 peptide-based vaccination are restricted to a certain HLA haplotype. We determined the frequency of HER-2 expression using the HercepTest for immunohistochemistry and HER-2 gene amplification by fluorescence in situ hybridisation (FISH) assay in oesophageal SCC (n=66). HER-2-positive tumours (1+/2+/3+) analysed by a HercepTest were observed in 30.3% of all the patients and HER-2 gene amplification evaluated by FISH was observed in 11.0% of all the patients, in which all HercepTest (3+) tumours were found to have gene amplification and three of six moderately positive (2+) tumours showed gene amplification. Furthermore, HER-2-positive cells were present more diffusely and were larger within each tumour in the patients who were HercepTest 3+ than those who were HercepTest 1+. Moreover, the survival rate in HER-2-positive group was significantly worse than that in HER-2-negative group. Also, the survival rate in the patients with HER-2 gene amplification was significantly worse than that without HER-2 gene amplification. In addition, oesophageal SCC patients with both HLA-A24-positive and HER-2-positive tumours (1+/2+/3+) accounted for 26% of these cases, and both HLA-A2- and HER-2-positive tumours accounted for 18% of them.     

宫颈腺癌HER-2/neu和P53的表达及其临床病理学意义

目的:研究宫颈腺癌HER-2/neu和P53蛋白的表达及其临床病理意义,探讨这些生物标记在临床治疗选择中的作用。方法:应用EnVision 免疫组织化学方法,对HER-2/neu、P53以及雌激素受体(ER)和孕激素受体(PR)蛋白在76例宫颈腺癌中的表达情况进行观察,并对其与宫颈腺癌临床病理特征的关系进行统计学分析。结果:HER-2/neu、P53、ER和PR蛋白在宫颈腺癌中的阳性表达率分别为18·4%(14/76)、36·8%(28/76)、27·6%(21/76)和25·0%(19/76)。HER-2/neu的过度表达与手术分期、肿瘤大小、组织学类型和分级及淋巴血管浸润等临床病理学特征无明显相关性;P53蛋白在高、中分化组(G1 G2)的阳性表达率显著低于低分化组(G3)(χ2=8·732,P=0·003),在有淋巴血管浸润的宫颈腺癌的表达率明显高于无淋巴血管浸润者(P=0·039);HER-2/neu与P53的过度表达呈正相关(r=0·27,P=0·018)。结论:HER-2/neu和P53蛋白过度表达可作为部分宫颈腺癌浸润和不良预后的良好标记物,对选择有效的治疗方案有重要意义。     

Evaluation of HER-2/neu gene amplification and protein expression in non-small cell lung carcinomas

HER-2/neu gene amplification and cell surface overexpression are important factors in breast cancer for prognosis and prediction of sensitivity to anti-HER-2/neu monoclonal antibody therapy. In lung cancer, the clinical significance of HER-2/neu expression is currently under evaluation. We investigated 238 non-small lung carcinomas for HER-2/neu protein overexpression by immunohistochemistry using the HercepTest. We found 2+ or 3+ overexpression in 39 patients (16%), including 35% in adenocarcinomas and 20% in large cell carcinomas, but only 1% of squamous cell carcinomas. Marked (3+) overexpression was uncommon (4%). The association between protein expression and gene copy number per cell, as determined by fluorescence in situ hybridisation assay, was investigated in 51 of these NSCLC tumours. Twenty-seven tumours (53%) were negative by both tests. Marked (3+) protein expression and gene amplification were present in only 4% of samples. In 11 tumours (21%), gene gain was accompanied by chromosomal aneusomy and did not result in high protein levels while in 7 (14%) the score 2+ was associated with maximum number of signals per cell <9. The prognostic implication of HER-2/neu protein expression was studied in 187 surgically resected tumours. No statistical difference in survival was observed comparing patients with positive (2+/3+) and negative tumours (0/1+), although 3+ patients showed a tendency to shorter survival. The therapeutic implications of protein expression and gene amplification in lung cancer need to be examined in prospective clinical trials.     

食管癌组织中HER-2/neu蛋白表达和基因扩增的检测及其临床意义

目的:检测食管癌组织中HER-2/neu蛋白表达及其基因扩增情况,并探讨其与食管癌患者临床病理指标的关系。方法:回顾性分析2000～2006年武汉大学人民医院收治的167例食管癌患者的临床资料。采用免疫组织化学和荧光原位杂交方法检测167例食管癌患者癌组织中HER-2/neu蛋白表达及基因扩增情况。结果:食管癌组织中HER-2/neu蛋白表达水平与基因扩增存在一定相关性(P<0.05),HER-2/neu蛋白的表达与食管癌分化程度和临床分期有关系(P<0.05),而HER-2/neu基因扩增仅与食管癌临床分期有关系(p<0.05)。HER-2/neu蛋白过表达和基因扩增阳性均可缩短食管癌患者的生存期(P<0.05)。结论:HER-2/neu蛋白过表达及基因扩增可能是食管癌患者的一个预后指标,联合检测HER-2/neu蛋白表达水平及基因扩增程度对于食管癌的靶向治疗可能具有一定指导意义。     

甲状腺癌HER—2／neu癌蛋白和mRNA表达的研究

「目的」观察人类甲状腺癌中ＨＥＲ－２／ｎｅｕ癌蛋白和ｍＲＮＡ的表达及其关系。「方法」应用免疫组化和同位素点杂交技术对新鲜冰冻甲状腺组织ＨＥＲ－２／ｎｅｕ癌蛋白和ｍＲＮＡ的表达进行研究。「结果」２１例甲状腺乳头状癌中１４例ＨＥＲ－２ｅｎｕ癌蛋白表达阳性,而其他类型肿瘤和非肿瘤组织均未见ＨＥＲ－２／ｎｅｕ ｍＲＮＡ表达研究显示,乳头状癌和淋巴结转移灶中,ｍＲＮＡ的表达水平明显高于其他肿瘤和非肿瘤组织。本研究还证明石蜡包埋的乳头状癌标本中ＨＥＲ－２／ｎｅｕ癌蛋白表达率明显低于新鲜标本。「结论」ＨＥＲ－２／ｎｅｕ蛋白的表达是甲状腺乳头状癌的一个特征性改变,ＨＥＲ－２ｎｅｕ蛋白的表达与ｍＲＮＡ的表达相一致,ＨＥＲ－２ｎｅｕ蛋白检测的稳定性取决于组织标本的质量。     

HER-2/neu amplification detected by fluorescence in situ hybridization in fine needle aspirates from primary breast cancer.

BACKGROUND: The HER-2/neu gene is amplified in 20-30% of human breast cancers and has been shown to have prognostic and predictive value for treatment with chemotherapy, hormone therapy and antibodies against the HER-2/neu domain (trastuzumab). The aim of our study was to evaluate the reliability of HER-2/neu determination by fluorescence in situ hybridization (FISH) on fine-needle aspirates (FNAs) from primary breast cancer patients by comparison with the results obtained by FISH and immunohistochemistry (IHC) on the corresponding histological sections. MATERIALS AND METHODS: HER-2/neu amplification was determined by FISH on 66 breast cancer FNAs. Twenty-three and 36 corresponding formalin-fixed, paraffin-embedded sections were assayed by FISH and by IHC, respectively, in order to detect HER-2/neu amplification and HER-2/neu protein expression. RESULTS: Twenty-seven per cent (18/66) of breast cancer FNAs showed amplification of HER-2/neu by FISH. Paired results by FISH cytology and FISH histology were available in 22 cases. Concordance was 91% (20/22). Paired results by FISH cytology and IHC were available in 36 cases. Concordance was 92% (33/36). Eighteen of 66 breast cancer FNAs were also submitted to flow cytometric DNA analysis. None of the diploid cases showed HER-2/neu amplification by FISH. Six out of the eight aneuploid cases were amplified and two were polysomic. CONCLUSIONS: HER-2/neu gene amplification can be reliably estimated by FISH on breast cancer FNAs and a good correlation has been found between FISH and IHC results from the corresponding histological sections.