乳腺癌可溶性HER-2/neu表达水平及其临床意义

目的 分析乳腺癌血清可溶性HER-2／neu水平并探讨其临床意义。方法 用ELISA方法分析105例乳腺癌患者、24例乳腺良性疾病及30名健康体检者血清可溶性HER-2／neu水平，并用免疫组化（IHC）方法分析乳腺癌组织切片中癌细胞的着色情况。结果 乳腺癌患者血清中可溶性HER-2／neu水平与正常对照和良性乳腺疾病相比，差异有统计学意义（P〈0．05）。随病程进展乳腺癌患者可溶性HER-2／neu的阳性率逐步升高，Ⅲ＋Ⅳ期的阳性率（63．3％）与Ⅰ期（7．7％）或Ⅱ期阳性率（14．5％）相比，差异有统计学意义（P〈0．05）；发生转移的乳腺癌患者可溶性HER-2／neu的阳性率（37．5％）与未转移患者（6．1％）相比，差异有统计学意义（P〈0．05）。血清HER-2／neu水平与IHC结果有显著正相关（P〈0．05）。结论 检测血清可溶性HER-2／neu水平可弥补免疫组化的不足，在乳腺癌监测随访、疗效观察尤其是用Hereeptin治疗后的疗效观察中发挥其独到的作用。     

乳腺癌患者血清HER-2／neu浓度及临床意义的分析

目的分析乳腺癌患者血清人表皮生长因子受体．2（HER-2／neu）的水平与组织HER-2表达水平的差异,分析血清HER-2／neul的影响因素以及与化疗疗效的相关性。方法采用酶联免疫吸附（ELISA）方法检测37例可手术的早期乳腺癌患者及74例晚期转移性乳腺癌患者血清HER-2／neu的水平,分析血清HER-2／neu与组织中HER-2的表达、临床参数的相关性。结果晚期乳腺癌患者血清HER-2／neu的水平与组织HER-2的状态一致性较好（P〈0．05）,早期乳腺癌患者中两者则无明显关系。乳腺癌血清HER-2的水平与血清CA153、CEA均无明显相关性。晚期乳腺癌血清HER-2的水平与肿瘤大小、远处转移数目、脉管瘤栓具有明显关系,而与月经状况、ER、PR均无明显关系（P〉0．05）。血清HER-2水平的变化与化疔疗效有关（P〈0．05）。结论晚期乳腺癌患者血清HER-2／neu水平与肿瘤组织中HER-2表达是一致的且与肿瘤负荷密切相关,可作为组织学检测的重要补充方法。     

Her-2／neu和CD138在乳腺癌中表达及意义

目的检测乳腺癌组织中Her-2／neu和CD138的表达情况,探讨二者在乳腺癌中表达的意义和相互关系。方法应用免疫组化方法（S-P法）观察102例乳腺癌,30例乳腺良性病变组织（15例乳腺良性肿瘤与15例正常乳腺组织）Her-2／neu和CD138的表达。结果①102例乳腺癌组织和30例乳腺良性病变组织中Her-2／neu阳性表达率分别为58％和0％,CD138阳性表达率分别为41％-42％和100％。乳腺癌组织中Her-2／neu阳性表达率显著高于其在良性病变组织的阳性表达率（P〈0．01）,良性病变组织中CD138阳性表达率显著高于乳腺癌组织（P〈0．015）。②Her-2／neu的表达与乳腺癌的淋巴结转移、临床分期密切相关（P〈o．05）。③CD138表达与乳腺癌的淋巴结转移、临床分期呈负相关（P〈0．05）。结论乳腺癌组织中Her-2／neu阳性表达与CD138的阴性表达显著相关（P〈O．01）。     

HER-2/neu在上皮性卵巢癌中的表达及其临床意义

目的：研究HER-2／neu在上皮性卵巢癌患者中的表达情况及其临床意义。方法：应用免疫组织化学SP法检测52例上皮性卵巢癌、10例卵巢交界性肿瘤、10例卵巢良性上皮性肿瘤中HER-2蛋白的表达情况。结果：上皮性卵巢癌中的HER-2阳性表达率显著高于卵巢良性上皮性肿瘤及卵巢交界性肿瘤，差异有显著性（P=0．045）。HER-2／neu蛋白在不同组织学类型、病理分级及临床分期上皮性卵巢癌中的表达差异无显著性（P〉0．05）。HER-2／neu蛋白阳性组平均生存时间短于阴性组（16．9个月vs40．7个月），经Log Rank检验，差异有显著性（P=0．005）。结论：HER-2／neu蛋白的过表达可能在上皮性卵巢癌的发生和发展中起一定作用，且与上皮性卵巢癌患者的不良预后相关．显著缩短患者术后生存时间。是上皮性卵巢癌患者的不良预后因素。[     

乳腺癌患者血液HER-2浓度与组织HER-2表达水平的一致性研究

目的探讨复发转移乳腺癌患者血清HER-2浓度与组织HER-2表达水平的一致性。方法应用ELISA法测定血清HER-2浓度;应用免疫组化(IHC)法检测乳腺癌组织HER-2表达水平;对检测结果进行统计分析。结果组织HER-2阳性的复发转移乳腺癌患者中有62例血清HER-2阳性,阳性符合率为74.7%,组织HER-2阴性的患者有96例血清HER-2为阴性,阴性符合率为82.1%。血清HER-2水平与组织HER-2表达状态呈正相关(kappa=0.653,P<0.01)。结论复发转移乳腺癌患者血清HER-2浓度与组织HER-2表达水平无显著差别,两种检测方法具有较好的一致性,血清HER-2浓度可作为临床个体化治疗的参考依据。     

三羟异黄酮对HER-2／neu过度表达乳腺癌裸鼠移植瘤血管生成的抑制作用

背景：血管生成是判断肿瘤预后的一个重要标志。原癌基因HER-2／neu高表达乳腺癌患者预后不良，大量研究资料表明三羟异黄酮对乳腺癌有抗癌作用。目的：探讨乳腺癌HER-2／neu表达状态与血管生成的关系，以及三羟异黄酮对HER-2／neu过度表达乳腺癌血管生成的影响。设计：以实验动物为研究对象，随机对照观察研究。单位：一所军医大学营养与食品卫生学教研室。材料：20只健康雌性BALB／c裸小鼠，SPF级，体质量(10&;#177;2)g，鼠龄3—4周，购自第三军医大学实验动物中心。方法：实验于2001—06／2002—03在第三军医大学营养与食品卫生学教研室完成。HER-2／neu表达阴性的乳腺癌MCF-7细胞通过转染HER-2／neu基因建立HER-2／neu过度表达的MCF-7细胞(命名为MCF-7／HER-2细胞)，建立MCF-7和MCF-7／HER-2细胞BALB／c裸鼠移植瘤模型。4周后，MCF-7／HER-2细胞移植瘤裸鼠随机分为3组：对照组，三羟异黄酮处理组和抗HER-2／neu抗体处理组。3周后处死动物，切下移植瘤，检测肿瘤体积、微血管密度和血管内皮生长因子表达。主要观察指标：主要结局：裸鼠移植瘤微血管密度(micrmessel density，MVD)及血管内皮生长因子(vasular endothelial growth factor，VEGF）。次要结局：①MCF-7细胞转染HER-2／neu的鉴定。②裸鼠移植瘤体积结果：MCF-7细胞移植瘤组、MCF-7／HER-2细胞移植瘤对照组、三羟异黄酮处理组、HER-2／neu抗体处理组微血管密度分别为(16&;#177;6)，(98&;#177;21)，(56&;#177;18)，(52&;#177;19)个／视野。MCF-7／HER-2细胞移植瘤的微血管密度和血管内皮生长因子水平显著高于MCF-7细胞移植瘤，而三羟异黄酮和抗HER-2／neu抗体处理明显降低MCF-7／HER-2细胞移植瘤的微血管密度和血管内皮生长因子水平，移植瘤体积变化特点与微血管密度和血管内皮生长因子水平变化类似。结论：乳腺癌过度表达HER-2／neu促进血管生成，而三羟异黄酮能抑制HER-2／neu过度表达乳腺癌的血管生成，这有助于改善乳腺癌的预后。     

抑制素和激活素在上皮性卵巢癌中的表达及其与卵巢癌发病机制的关系

目的：研究人表皮生长因子受体2（human epidermal growth factor receptor 2，HER-2/neu）在上皮性卵巢癌患者中的表达情况及其临床意义。方法：应用免疫组织化学SP法检测52例卵巢癌、10例交界性卵巢肿瘤、10例良性上皮性卵巢肿瘤中HER-2蛋白的表达情况。结果：（1）卵巢癌中的HER-2阳性表达率显著高于卵巢良性上皮性肿瘤及卵巢交界性肿瘤，差异有显著性(P=0.045)。（2）HER-2/neu蛋白在不同组织学类型、病理分级及临床分期卵巢癌中的表达差异无显著性(P>0.05)。（3）生存时间比较：HER-2/neu蛋白阳性组平均生存时间短于阴性组（16.9月与40.7月），经Log Rank检验，差异有显著性（P=0.005）。结论：HER-2/neu蛋白的过表达可能在卵巢癌的发生和发展中起一定作用，且与卵巢癌患者的不良预后相关，显著缩短患者术后生存时间，是卵巢上皮性癌患者的不良预后因素。     

COX-2、HER-2和VEGF在乳腺癌中的表达及其临床意义

【目的】研究环氧化酶-2（cOx-2）在乳腺癌组织中的表达情况与,临床病理指标及预后之间的相关性及与VEGF和HER-2表达的关系。【方法】检测120例乳腺癌石蜡包埋组织中COX-2、HER-2和VEGF的表达。【结果】120例中cox-2的高表达率为38．3％（46／120）。淋巴结转移阳性组和阴性组以及不同临床分期的病人之间COX-2的表达差异有显著意义（P〈0．05）,而与病人年龄、肿瘤大小、病理类型、ER、PR等差异无显著意义（P〉0．05）。COX-2在乳腺癌组织中的表达与HER-2／neu、VEGF的表达密切相关（P〈0．01）。COX-2和HER-2共同表达的患者与两者单独表达的患者相比,前者的TNM分期较晚,有显著差异（P=0．027）。COx_2和VEGF共同表达与单独表达的TNM分期无显著差异（P=0．479）。COX-2低表达的患者总生存期较长,与高表达患者的总生存期有显著差异（P〈0．005）。【结论】乳腺癌组织中COX-2的高表达可提示恶性程度高和预后不良。乳腺癌组织中COX-2与HER-2、VEGF的表达相关,且COX-2和HER-2／neu同时高表达时预后更差。     

乳腺癌患者HER-2胞外结构域的临床价值

目的探讨乳腺癌患者血清人类表皮生长因子受体2(HER-2)胞外结构域(ECD)的临床价值。方法选取乳腺癌患者93例、良性乳腺病患者40例及健康女性(正常对照组)55名,采用化学发光法定量检测所有对象血清HER-2 ECD水平,并分析乳腺癌患者血清HER-2 ECD水平与癌组织HER-2表达状态的一致性、血清HER-2 ECD与乳腺癌临床病理特征的关系及其对乳腺癌的诊疗效能。结果乳腺癌组血清HER-2 ECD水平[16.00(12.30,20.10)ng/mL]明显高于良性乳腺病组[6.05(4.85,7.95)ng/mL,P0.05]和正常对照组[5.30(4.00,6.80)ng/mL,P0.05]。乳腺癌患者癌组织HER-2表达阳性者的血清HER-2 ECD阳性率为73.08%,血清HER-2 ECD与癌组织HER-2表达的一致性较好。不同组织学分级、有无淋巴结转移及不同临床分期的乳腺癌患者血清HER-2 ECD水平差异有统计学意义(P0.05)。结论乳腺癌患者血清HER-2 ECD与乳腺癌的临床特征关系密切,有望成为早期诊断乳腺癌、监测疗效、判断预后的重要指标之一。     

HER-2/neu基因表达与卵巢上皮性癌的预后

【目的】探讨HER-2／neu基因在卵巢上皮性癌中的表达水平及其与预后的关系。【方法】利用免疫组化检测93例卵巢上皮性癌组织中HER-2／neu基因表达水平,并结合临床病理及随访资料进行分析。【结果】HER-2／neu基因主要为细胞膜及胞浆表达,阳性率为44．1％。其表达与临床分期、病理类型、组织学分级及残余灶直径等无相关性;但与预后相关。HER-2／neu表达阴性组的1,3,5年生存率分别为90．4％、74．0％、54．6％,弱阳性组分别为81．1％、45．4％、27．9％,强阳性组分别为80．0％、20．0％、10．0％。差异有显著性(P=0．0028)。COX模型多因素分析提示：HER-2／neu表达水平是影响卵巢癌预后的独立因素(P=0．005)。【结论】HER-2／neu基因表达水平可作为预测卵巢上皮性癌预后的可靠指标。     

Serum HER-2/neu in the management of breast cancer patients

The clinical role of HER-2/neu, a 185 kD epithelial transmembranous protein, has evolved after the approval of the anti-HER-2/neu targeted monoclonal antibody trastuzumab (Herceptin) for the therapy of metastatic breast cancer. The extracellular domain of HER-2/neu undergoes proteolytic cleavage from the full-length protein by metalloproteases, and is shed into the blood as a circulating antigen. While HER-2/neu gene amplification and/or protein overexpression are detected in approximately 25% of primary breast cancers, serum HER-2/neu levels are elevated beyond the upper limit of normal in 50 to 60% of stage IV breast cancer patients. HER-2/neu in serum can be detected by enzyme immunoassays (manual and automated versions). It has been shown to have prognostic and predictive information in breast cancer patients. Monitoring for recurrence by serum HER-2/neu reaches a high sensitivity for HER-2/neu positive tumors. Longitudinal follow-up of patients during any kind of systemic therapy allows for monitoring of the therapeutic success. When utilized in these applications, serum HER-2/neu testing is complementary to HER-2/neu tissue results and to the determination of classical tumor markers such as CA 15-3, CA 27.29 and CEA, which are not targeted by specific forms of systemic therapy.     

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.     

Relationship of serum HER-2/neu and serum CA 15-3 in patients with metastatic breast cancer

BACKGROUND: Serum HER-2/neu antigen concentrations have been reported to correlate with increased tumor volume in patients with breast cancer. We measured serum CA 15-3, a surrogate marker of disease burden, and correlated serum CA 15-3 with serum HER-2/neu and analyzed the association of both markers with clinical outcomes. METHODS: Pretreatment serum samples from 566 patients were retrospectively analyzed from 2 phase III clinical trials of estrogen receptor-positive (ER(+)), ER(-)/progesterone receptor-positive, or ER status unknown metastatic breast cancer patients randomized in two similar studies to receive second-line hormone therapy with either megestrol acetate or an aromatase inhibitor (fadrozole). The extracellular domain of the HER-2/neu (c-erbB-2) oncogene and serum CA 15-3 were measured by ELISA on the Bayer Immuno 1. RESULTS: Serum HER-2/neu protein was increased in 168 patients (30%), and CA 15-3 was increased in 337 (60%) patients. Serum CA 15-3 and HER-2/neu were weakly correlated (r = 0.39; P <0.0001). The clinical benefit (complete responses plus partial responses plus stable disease) of endocrine therapy was significantly lower in patients with increased serum HER-2/neu. When adjusted for serum HER-2/neu, serum CA 15-3 was not predictive of response rates. The median time to progression was shorter in patients with increased serum HER-2/neu (89 days) compared with patients with normal serum HER-2/neu (176 days). Survival was significantly shorter in patients with increased serum HER-2/neu (513 vs 869 days; P <0.0001) or increased serum CA 15-3 (689 vs 939 days; P <0.0001). This observation was confirmed by multivariate analysis. CONCLUSIONS: Serum HER-2/neu is a significant independent predictive and prognostic factor in hormone receptor-positive metastatic breast cancer, even when adjusted for tumor burden as measured by CA 15-3. The combination of increased serum HER-2/neu and increased serum CA 15-3 predicts a worse prognosis than does increased CA 15-3 alone.     

AE37: a novel T-cell-eliciting vaccine for breast cancer

INTRODUCTION: Immunotherapy, including vaccines targeting the human EGFR2 (HER-2/neu) protein, is an active area of investigation in combatting breast cancer. Several vaccines are currently undergoing clinical trials, most of which are CD8(+) T-cell-eliciting vaccines. AE37 is a promising primarily CD4(+) T-cell-eliciting HER-2/neu breast cancer vaccine currently in clinical trials. AREAS COVERED: This article reviews preclinical investigations as well as findings from completed and ongoing Phase I and Phase II clinical trials of the AE37 vaccine. EXPERT OPINION: Clinical trials have shown the AE37 vaccine to be safe and capable of generating peptide-specific, durable immune responses. This has been shown in patients with any level of HER-2/neu expression. Early clinical findings suggest there may be benefit to AE37 vaccination in preventing breast cancer recurrence.     

Level of HER-2/neu protein expression in breast cancer may affect the development of endogenous HER-2/neu-specific immunity

We questioned whether the incidence or magnitude of the humoral or cellular immune response to the self-tumor antigen HER-2/neu is influenced by the level of HER-2/neu protein overexpression as defined by immunohistochemical staining of tumors in breast cancer patients. We obtained peripheral blood from 104 women with stage II, III, and IV pathologically confirmed HER-2/neu-overexpressing breast cancer. Patients were categorized with +1 (n = 14), +2 (n = 20), or +3 (n = 70) HER-2/neu overexpression by institutional pathologic report. Circulating antibodies to HER-2/neu were evaluated using ELISA. T-cell responses to HER-2/neu were measured using an antigen-specific tritiated thymidine incorporation assay. Eighty-two percent of subjects with HER-2/neu antibodies were +3 overexpressors compared with 18% +2 overexpressors and 0% +1 overexpressors, a highly significant difference (P < 0.001), and there were significant differences in the magnitude of the HER-2/neu-specific antibodies between groups with varying HER-2/neu protein expression (P = 0.022). In addition, 65% of subjects with HER-2/neu-specific T cells were +3 overexpressors compared with 16% +2 overexpressors and 19% +1 overexpressors (P = 0.001). Data presented here indicate that endogenous HER-2/neu-specific humoral and T-cell immunity is greater in patients with +3 protein overexpression in their tumors than in patients with lower levels of HER-2/neu expression. Overexpression of a self-tumor-associated protein is a potential mechanism by which immunogenicity is enhanced and may aid in the identification of biologically relevant proteins to target for immune-based molecular cancer therapies.     

HER-2/neu gene copy number quantified by real-time PCR: comparison of gene amplification,heterozygosity, and immunohistochemical status in breast cancer tissue

BACKGROUND: Amplification of the oncogene HER-2/neu influences breast cancer pathogenesis, and therapy and prognosis may be affected by the degree of amplification. The extent of amplification or protein overexpression typically is analyzed by fluorescence in situ hybridization or immunohistochemistry (IHC), but quantitative PCR techniques have been described that may provide alternatives to these methods. METHODS: We developed a rapid-cycle, real-time PCR assay for quantification of HER-2/neu gene status. We compared results obtained with this assay with short tandem repeat findings by capillary electrophoresis (CE) and with protein overexpression assessments by IHC. Accuracy and linearity were tested on cell lines and with simulation experiments. We analyzed the amplification of HER-2/neu in 51 clinical tissue samples from patients with suspected breast cancer. RESULTS: The intra- and interrun CVs for HER-2/neu quantification by real-time PCR were 12% and 18%, and the CV for different simulated amplification and deletion experiments was <7%. The results for HER-2/neu gene status in cell lines matched the values reported in literature. We detected HER-2/neu amplification by real-time PCR in 11 samples, all from patients with invasive ductal carcinoma. Allelic imbalances were found by CE analyses in three samples and by protein overexpression in six samples; five of these were also detected by real-time PCR. Comparison of the quantification results with known prognostic indices yielded results similar to those reported in several other published studies. CONCLUSIONS: The assay is suitable for accurate and precise quantification of HER-2/neu copy numbers in tumor tissue samples obtained in routine clinical practice.     

HER-2 protein concentrations in breast cancer cells increase before immunohistochemical and fluorescence in situ hybridization analysis turn positive.

BACKGROUND: The level of HER-2/neu in breast cancer cells is normally measured by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH). It determines whether patients should be treated with trastuzumab (Herceptin). In this study, HER-2 protein in breast cancer tissue was measured using a quantitative method. METHODS: Tissue samples of malignant and adjacent benign breast tissue were collected from 118 consecutive women admitted for surgical treatment of breast cancer. The HER-2 protein concentration was determined by 2 HER-2 assays: ELISA and the Bayer ADVIA Centaur assay. Paraffin-embedded tissue sections of the corresponding tumors were analyzed by IHC and FISH. RESULTS: Increased HER-2 concentrations in cancer tissue were found compared to autologous reference tissue (p<0.0001, Wilcoxon test) and normal breast tissue (p<0.0001, Mann-Whitney test). Good concordance rates were observed between the methods: 95.8% for IHC and FISH; 86.4% for IHC and ELISA; and 87.3% for FISH and ELISA. The HER-2 positivity rate was determined to 26.3% by ELISA, 12.7% by IHC and 16.9% by FISH. No correlation was found with tumor grade, axillary node status or serum HER-2 levels. CONCLUSIONS: Detection of HER-2 overexpression by measuring HER-2 in tissue extracts by ELISA seems to be more sensitive than IHC and FISH. This suggests that some patients deprived of Herceptin treatment may benefit from this treatment and may also explain the conversion phenomenon from HER-2-negative to HER-2-positive observed in relapse and metastatic disease.