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。     

复发转移乳腺癌患者血清HER2水平与组织表达状态的相关性

目的探讨复发转移乳腺癌患者血清HER2水平与组织表达状态间的联系,分析血清HER2检测是否可作为组织HER2检测的补充手段。方法采用酶联免疫吸附试验(ELISA)对72例女性复发转移乳腺癌(组织HER2阳性组30例,组织HER2阴性组42例)和30例健康体检女性(健康对照组)进行血清HER2水平检测,将各组的血清HER2水平进行比较,并与组织HER2表达状态进行对比分析。结果组织HER2阳性组的血清HER2水平显著高于组织HER2阴性组和健康对照组,差异有统计学意义(P<0.0001);组织HER2阴性组血清HER2水平与健康对照组比较差异无统计学意义(P=0.163);组织HER2阳性组有22例血清HER2阳性,阳性符合率为73.4%(22/30),组织HER2阴性组有32例血清HER2阴性,阴性符合率为78.6%(32/42)。血清HER2水平与组织HER2表达状态呈正相关(χ2=21.626,r=0.547)。结论血清HER2水平与组织HER2表达状态呈正相关,且血清HER2可以作为组织HER2检测的补充手段。     

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。     

有丝分裂原活化蛋白激酶对HER-2阳性乳腺癌化疗疗效考核价值

目的 探讨有丝分裂原活化蛋白激酶(MAPK)对HER-2阳性乳腺癌患者化疗的疗效及生存预后的预测价值。方法 选择2010年5月～2012年5月本院收治的83例HER-2阳性乳腺癌患者为研究对象,所有患者均接受新辅助化疗和手术治疗,采用免疫组化染色检测癌组织和癌旁组织MAPK、MAPK磷酸化(pMAPK)蛋白表达,采用RT-PCR检测组织MAPK、pMAPK mRNA表达;以患者年龄、手术方式、临床分期、月经、雌激素受体、MAPK、pMAPK表达等作为观察指标,分析影响乳腺癌化疗疗效的因素。结果 癌组织MAPK、pMAPK mRNA及蛋白表达均显著高于癌旁组织(P0.05)。83例乳腺癌患者CR 15例,占18.1%;PR 45例,占54.2%;SD 22例,占26.5%;PD 1例,占1.2%.临床有效共计60例占72.3%。单因素分析显示MAPK、pMAPK蛋白阳性患者临床有效率显著低于MAPK、pMAPK蛋白阴性患者(P0.05);多因素分析显示MAPK、pMAPK表达是影响乳腺癌化疗疗效的独立危险因素(P均0.05)。MAPK阳性患者3年无复发生存率为41.8%,MAPK阴性患者为60.7%,MAPK阳性3年无复发生存率显著低于MAPK阴性(χ~2=11.037,P0.001);pMAPK阳性患者3年无复发生存率为39.1%,pMAPK阴性患者为58.7%,pMAPK阳性3年无复发生存率显著低于MAPK阴性(χ~2=4.103,P=0.041)。结论 MAPK、pMAPK可以作为HER-2阳性乳腺癌患者化疗疗效及生存预后的评估指标之一。     

荧光原位杂交法与免疫组织化学法检测乳腺癌HER-2基因扩增及蛋白表达的对比研究

目的 比较桂林市荧光原位杂交法(FISH)和免疫组织化学法(IHC)检测乳腺癌组织中人表皮生长因子受体(HER-2)基因扩增及其蛋白表达情况的一致性,探讨FISH与IHC检测乳腺癌HER-2基因状态的临床意义.方法 应用IHC和FISH对50例乳腺癌患者HER-2蛋白表达、基因扩增情况及17号染色体倍体性进行检测,分析IHC与FISH检测HER-2基因状态的差异以及HER-2蛋白状态与17号染色体倍体性的相天性.结果 FISH与IHC结果 总的符合率为82.0%,两者之间存在较好的一致性(kappa=0.640),FISH检测IHC结果 为3+、2+和0/1+者的符合率分别为92.9%、70.0%和80.8%;IHC3+及2+者出现17号染色体多体性的几率比IHC0/1+者高(P<0.05).结论 FISH可以准确和稳定地检测乳腺癌组织中HER-2的基因状态及17号染色体倍体性;FISH检测IHC3+者符合率较高,FSH与IHC的差异主要在于IHC2+和IHC0/+组,IHC仅作为检测HER-2基因状态的初筛方法 ,而IHC结果 为2+或0/1+者的HER-2基因状态必须应用FISH进一步确定.     

乳腺癌组织中HER-2、VEGF和Ki67的表达及临床意义

目的探讨人表皮生长因子受体-2(HER-2)、血管内皮生长因子(VEGF)和Ki67在乳腺癌组织中的表达及其与临床病理因素的关系。方法采用免疫组化法检测49例乳腺癌患者组织中HER-2、VEGF和Ki67的表达。结果 HER-2、VEGF和Ki67在乳腺癌组织中的阳性表达率分别为36.7%、69.4%、79.6%;HER-2在乳腺癌组织学分级和淋巴结转移中的表达差异有统计学意义(P<0.05);VEGF在乳腺癌淋巴结转移中的表达有显著差异(P<0.05);Ki67表达在乳腺癌组织学分级中的差异有统计学意义(P<0.05)。结论 HER-2、VEGF和Ki67与乳腺癌的发生、发展密切相关,联合检测可作为判定乳腺癌预后的评估指标。     

乳腺癌患者三阴性与HER-2过表达的临床病理特征及预后比较

目的探讨乳腺癌患者三阴性与人表皮生长因子受体2(HER-2)过表达的临床病理特征及预后。方法采用免疫组织化学染色法检测568例乳腺癌组织中激素受体(ER)、孕激素受体(PR)、HER-2的表达。根据乳腺癌分子表型不同进行分型,三阴性(ER、PR、HER-2均为阴性),HER-2过表达型(ER与PR阴性,HER-2阳性),对两组患者的临床病理资料进行分析。采用电话或门诊随访患者至2013年12月年或患者死亡,比较两组患者2年及5年无病生存率与总生存率、无病生存时间、复发或转移情况,采用Kaplan-Meier法进行生存分析,log-rank检验进行生存率比较。结果 568例患者中,三阴性与HER-2过表达型分别为85例(14.96%)、12例(21.13%)。三阴性组恶性肿瘤家族史患者比率、组织学分级3级患者比率和浸润性导管癌患者比率均显著高于HER-2过表达组,差异具有显著性(P0.05)。患者最长随访时间为120个月,最短为10个月,平均(85.89±38.59)个月。三阴性组乳腺癌复发或转移率显著高于HER-2过表达者(P0.05),而且脑转移发生率较高,而HER-2过表达组则以骨转移为主。三阴性乳腺癌无病生存时间明显短于为HER-2过表达组(47.99±24.59 vs.75.62±59.5)个月,两组差异有统计学意义(P0.05)。三阴性组2年及5年无病生存率及总生存率均低于HER-2过表达组,但两组差异不显著(P0.05)。结论三阴性与HER-2过表达乳腺癌相比,恶性肿瘤家族史比率及肿瘤恶性度高,更易发生脑转移,预后相对较差。     

HER-2基因表达在乳腺原发癌和淋巴结转移癌发病中的作用研究

目的探讨使用荧光原位杂交技术检测乳腺原发癌和淋巴结转移癌HER-2基因的表达及意义。方法选取海口市人民医院2013年1月至2014年1月收治的乳腺原发癌患者40例作为研究对象,对其临床资料进行回顾性分析。应用荧光原位杂交技术(FISH)与免疫组织化学技术(IHC)对所有患者手术切除标本的HER-2基因表达予以检测。结果 FISH阳性率为42.5%,IHC为57.5%;IHC阴性时与FISH符合率为88.2%,IHC(+)与FISH符合率为37.5%,IHC(++)与FISH符合率为83.3%,IHC(+++)与FISH符合率为100.0%。淋巴结转移阳性率为66.67%,未转移阳性率为32.14%,对比差异有统计学意义(P0.05)。结论 IHC对HER-2基因予以检测时若结果为阴性或者强阳性则与FISH符合率较高,同时HER-2基因表达与腋淋巴结转移之间呈正相关关系。     

HER-2预测术后ER阴性乳腺癌患者长期生存的探讨

目的分析HER-2和ER表达对术后乳腺癌患者长期生存时间的预测作用。方法收集143例经长期随访（〉5年）的乳腺癌患者的组织标本，并通过免疫组化进行HER-2、ER检测，统计学分析这些指标与患者生存时间的关系。结果在143例具有完整治疗的乳腺癌患者中，Cox回归分析发现，ER和HER．2的表达状态明显与患者的预后有关。以ER表达作为分类变量，发现在ER阴性组，HER-2阳性患者的生存时间明显短于阴性患者。在生存曲线上，HER-2阴性患者生存80个月的可能性大于阳性患者。在ER阳性组，虽然HER-2阳性患者的生存时间与阴性患者存在差异，但无统计学意义。结论HER-2是预测术后ER阴性乳腺癌患者长期生存时间的理想生物学标记。     

PCR检测Ⅰ期HER-2阳性乳腺癌患者腋窝淋巴结HER-2表达的意义

[目的]探讨聚合酶链反应(PCR)检测对Ⅰ期人表皮生长因子受2(HER-2)阳性乳腺癌腋窝淋巴结H ER-2表达检出率的影响及其表达与腋窝淋巴结微转移与预后的关系.[方法]收集两院住院资料完整的128例女性Ⅰ期HER-2阳性乳腺癌患者的临床病理资料,采用PCR检测患者腋窝淋巴结HER-2表达并进行随访,用Kaplan-...     

胃癌组织HER-2基因扩增和蛋白表达检测及临床意义

目的 探讨免疫组化（immunohistochemistry,IHC）法检测胃癌HER-2蛋白表达和荧光原位杂交（fluorescence in situ hybridization,FISH）法检测HER-2基因扩增的临床意义.方法 收集256例胃癌患者手术切除及胃镜活检的胃癌组织标本,分别采用IHC法及FISH法检测胃癌组织HER-2蛋白表达及基因扩增状态,并对结果进行统计学分析.结果 256例胃癌组织标本中,有26例HER-2蛋白表达阳性,阳性率为10.16％;46例HER-2基因有扩增,阳性率17.97％.26例HER-2蛋白表达（3＋）的标本中,24例有HER-2基因扩增,2例无扩增;95例HER-2蛋白表达（2＋）的标本中,20例有基因扩增,75例无扩增;90例HER-2蛋白表达（1＋）的标本中,2例有基因扩增,88例无扩增;45例HER-2蛋白表达（0）的标本中,HER-2基因均无扩增.IHC法和FISH法检测HER-2表达阳性率差异无统计学意义（P＞0.05）.结论 IHC法检测胃癌HER-2蛋白表达可作为临床治疗的初筛,HER-2蛋白表达（2＋）的患者应常规行FISH法,以确定HER-2基因扩增状态.FISH法检测结果具有较高的稳定性和可靠性,联合IHC法可更准确和客观评价胃癌预后,并指导临床选用靶向药物治疗.     

Impact of polysomy 17 on HER-2/neu immunohistochemistry in breast carcinomas without HER-2/neu gene amplification

Her-2/neu, a proto-oncogene located on chromosome 17, is an important biomarker in breast carcinoma. Immunohistochemistry (IHC) is currently the most widely used method for assessing Her-2/neu status. Some IHC-positive cases do not show Her-2/neu gene amplification by fluorescence in situ hybridization (FISH). It has been suggested that some of these IHC “false positive” results may in part be due to increased copy number of chromosome 17 resulting in increased Her-2/neu protein expression. We analyzed IHC and FISH data from 561 cases of invasive breast carcinoma to test this hypothesis. IHC and FISH for Her-2/neu were performed on formalin-fixed, paraffin-embedded sections of 561 invasive breast carcinomas. The IHC results were interpreted as 0, 1+, 2+, or 3+ according to the manufacturer’s recommended criteria. The FISH results were expressed as a ratio of Her-2/neu/chromosome 17 and were interpreted as positive (> = 2.0) or negative (<2.0) for gene amplification according to the manufacturer’s recommended scoring system. We found that in IHC 3+/FISH-negative cases (n = 15) both the average chromosome 17 copy number and the average Her-2/neu copy number were significantly higher than that in IHC (0 to 2+)/FISH-negative cases (n = 411) (2.45 vs. 1.68; P < 0.0001, and 3.19 vs. 1.95; P < 0.0001, respectively). In contrast, the IHC 2+/FISH-negative cases did not exhibit a significantly increased number of chromosome 17 compared to IHC 0 to 1+ cases. In addition, the average copy number of chromosome 17 in FISH-positive cases (n = 135) was significantly higher than that in FISH-negative cases (n = 426) (2.27 vs. 1.70; P < 0.0001), indicating a general association of increased chromosome 17 copy number with Her-2/neu gene amplification. Thus, our data suggest that IHC 3+ immunostaining without scorable gene amplification may indeed be, at least in some cases, the result of increased Her-2/neu protein expression secondary to an increased copy number of chromosome 17, associated with an increased total number of Her-2/neu gene copies per tumor cell.     

HER-2/neu-gene engineered dendritic cell vaccine stimulates stronger HER-2/neu-specific immune responses compared to DNA vaccination

HER-2/neu is a candidate for developing breast cancer-targeted immunotherapeutics. Although DNA-based and HER-2/neu transgene-modified dendritic cell (DC)-based vaccines are potent at eliciting HER-2/neu-specific antitumor immunity, there has been no side-by-side study comparing them directly. The present study utilizes an in vivo murine tumor model expressing HER-2/neu antigen to compare the efficacy between adenovirus (AdVneu)-transfected dendritic cells (DC(neu)) and plasmid DNA (pcDNAneu) vaccine. Our data showed that DC(neu) upregulated the expression of immunologically important molecules and inflammatory cytokines and partially converted regulatory T (Tr)-cell suppression through interleukin-6 (IL-6) secretion. Vaccination of DC(neu) induced stronger HER-2/neu-specific humoral and cellular immune responses than DNA vaccination, which downregulated HER-2/neu expression and lysed HER-2/neu-positive tumor cells in vitro, respectively. In two HER-2/neu-expressing tumor models, DC(neu) completely protected mice from tumor cell challenge compared to partial or no protection observed in DNA-immunized mice. In addition, DC(neu) significantly delayed breast cancer development in transgenic mice in comparison to DNA vaccine (P<0.05). Taken together, we have demonstrated that HER-2/neu-gene-modified DC vaccine is more potent than DNA vaccine in both protective and preventive animal tumor models. Therefore, DCs genetically engineered to express tumor antigens such as HER-2/neu represent a new direction in DC vaccine of breast cancer.     

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.     

Immunization with a HER-2/neu helper peptide vaccine generates HER-2/neu CD8 T-cell immunity in cancer patients

CD4 T-cell help is required during the generation and maintenance of effective antitumor CD8 T cell-mediated immunity. The goal of this study was to determine whether HER-2/neu-specific CD8 T-cell immunity could be elicited using HER-2/neu-derived MHC class II "helper" peptides, which contain encompassed HLA-A2-binding motifs. Nineteen HLA-A2 patients with HER-2/neu-overexpressing cancers received a vaccine preparation consisting of putative HER-2/neu helper peptides p369-384, p688-703, and p971-984. Contained within these sequences are the HLA-A2-binding motifs p369-377, p689-697, and p971-979. After vaccination, the mean peptide-specific T-cell precursor frequency to the HLA-A2 peptides increased in the majority of patients. In addition, the peptide-specific T cells were able to lyse tumors. The responses were long-lived and detectable for more than 1 year after the final vaccination in select patients. These results demonstrate that HER-2/neu MHC class II epitopes containing encompassed MHC class I epitopes are able to induce long-lasting HER-2-specific IFN-gamma-producing CD8 T cells.     

Automated assay for HER-2/neu in serum

BACKGROUND: The extracellular domain of the HER-2/neu oncogene product is increased in sera of some patients with epithelial cancers. Our aim was to develop an automated serum assay for the extracellular domain of the HER-2/neu protein. METHODS: We used a monoclonal antibody labeled with fluorescein for capture and a monoclonal Fab' fragment labeled with alkaline phosphatase for detection. Separation of bound and free detection conjugate was performed with magnetizable particles coated with monoclonal antibody to fluorescein. Alkaline phosphatase activity was measured kinetically at 405 or 450 nm. RESULTS: The assay was linear from 0.1 to 250 microg/L. No hook effect was evident up to 10 000 microg/L. Within-run imprecision (CV) was 0.8-1.2%, and total imprecision was 1.1-1.7%. Cross-reactivity with human epidermal growth factor receptor, which has extensive homology with HER-2/neu extracellular domain, was <0.6%. Human anti-mouse antibodies, heterophilic antibodies, and rheumatoid factor did not interfere, nor did the therapeutic monoclonal antibody Herceptin((R)). In 51 healthy females, the mean value was 9.3 microg/L with a range of 6.4-14.0 microg/L. No reagent lot-to-lot variability was detected over four lots of reagents tested. CONCLUSION: The Bayer Immuno 1(TM) assay for HER-2/neu was precise and resistant to interferences, characteristics that are essential for longitudinal monitoring of cancer patients.     

Evaluation of the quantitative analytical methods real-time PCR for HER-2 gene quantification and ELISA of serum HER-2 protein and comparison with fluorescence in situ hybridization and immunohistochemistry for determining HER-2 status in breast cancer patients

BACKGROUND: HER-2 status is generally determined by immunohistochemistry (IHC) or fluorescence in situ hybridization (FISH). Both methods are only semiquantitative, require a tumor sample, and can be difficult to reproduce. We compared these methods with 2 quantitative approaches, one measuring HER-2 gene copy number in tissue by real-time quantitative PCR (qPCR), and the other measuring shed HER-2 protein in serum by ELISA in patients with metastatic disease. METHODS: We analyzed 52 cases of metastatic breast cancer for which both serum collected at the diagnosis of metastasis and stored primary breast tumor specimens were available. The within- and between-run imprecision of real-time qPCR and ELISA were evaluated according to Clinical and Laboratory Standards Institute (formerly known as NCCLS) recommendations. Concordance among the 4 methods was assessed by calculating the kappa statistic and its 95% confidence interval (95% CI). RESULTS: The CVs for within- and between-run imprecision were both <10% with qPCR and ELISA. There was good agreement of results between qPCR and IHC (kappa = 0.81; 95% CI, 0.64-0.99), qPCR and FISH (kappa = 0.77; 95% CI, 0.58-0.96), ELISA and IHC (kappa = 0.65; 95% CI, 0.41-0.89); and ELISA and FISH (kappa = 0.69; 95% CI, 0.46-0.92). CONCLUSIONS: Measurements of HER-2 gene expression by qPCR and of serum HER-2 protein by ELISA are highly reproducible approaches for determining HER-2 status in metastatic breast cancer. In addition, ELISA eliminates the need for biopsy.     

人表皮生长因子受体-2在胃癌组织中的表达及意义

目的探讨胃癌组织中人表皮生长因子受体-2(human epidermal growth factor receptor 2,HER-2)的表达及与临床病理特征的关系。方法应用免疫组化Envision二步法检测70例胃癌组织中HER-2的表达情况,并结合其临床病理特点进行分析。结果 HER-2在胃癌组织中的阳性表达率为27.1%(19/70),阳性过表达率为15.7%(11/70)。阳性表达与肿瘤分化程度、侵袭深度、有无淋巴结转移及TNM分期有关(P<0.05),而与患者的性别、年龄、肿瘤部位和大小无关(P>0.05)。结论 HER-2的表达参与胃癌的生长、侵袭和转移过程。检测HER-2有助于筛选对曲妥珠单抗(Herceptin)靶向治疗敏感的胃癌患者,也为胃癌的预后判断提供客观的参考指标。     

HER-2基因原位杂交方法学探讨

目的 尝试用新型自动化原位杂交仪(HS400Pro)建立规范化、标准化操作步骤.方法 可有效降低实验所需探针浓度、提高杂交效率.对5例已知HER-2阳性的乳腺癌存档蜡块,采用动态杂交技术行2种不同DNA探针标记(DIG和FITC),分别用手工和HS400Pro全自动杂交检测系统对比检测结果.结果 探针用专利杂交液按1∶40稀释后,采用新型自动化原位杂交仪37℃杂交12 h与手工37℃杂交18h结果一致.说明该仪器可以基本替代手工开展ISH.结论 自动化原位杂交仪的使用是实现ISH自动化、标准化、规范化的好方法.但要继续扩大检测样本量,找出HS400Pro的最佳杂交条件.