168例乳腺癌HER-2免疫组化与FISH技术检验分析

目的:比较免疫荧光原位杂交(FISH)检测乳腺癌组织中HER2基因扩增与免疫组织化学(IHC)染色检测HER2蛋白表达的一致性。方法:收集乳腺癌改良根治标本168例,所有病例均未行术前辅助化疗。10例正常乳腺组织为对照。结果:经统计学Pearson卡方检验表明FISH是乳腺癌HER2状态检测的金标准。结论:FISH与IHC两种方法具有很好的一致性,IHC可以用于乳腺癌HER2检测。     

浸润性乳腺癌HER-2基因扩增与蛋白表达的关系研究

目的 了解HER-2基因与蛋白在乳腺癌中表达状况,探讨HER-2基因与Ki-67、P53、ER、PR、年龄、淋巴结转移、TNM分期的相关性及其临床意义.方法 收集150例浸润性乳腺癌组织石蜡标本,用荧光原位杂交( FISH)技术和免疫组化(IHC)法分别检测标本中的HER-2基因及HER-2、Ki-67、P53、ER、PR蛋白表达情况.结果 150例浸润性乳腺癌标本HER-2基因扩增率为32.68％ (49/150);其中,在HER-2蛋白(+++)的病例中HER-2基因扩增率为90.32％ (28/31);HER-2蛋白(++)的病例中扩增率34.78％ (16/46);HER-2蛋白(+)和(-)的扩增率分别为11.54％(3/26)、4.25％(2/47).HER-2基因与ER、PR表达及TNM分期相关,而与年龄、Ki-67、P53、淋巴结转移无关.结论 FISH技术和IHC技术检测HER-2基因和蛋白表达在(+++)和(-)的病例中一致性较好,IHC可作为替代FISH检测HER-2基因状态的方法.     

FISH与IHC检测414例乳腺癌Her-2基因表达状态的比较

目的比较荧光原位杂交(FISH)和免疫组织化学(IHC)检测乳腺癌Her-2基因扩增情况的一致性与相关性。方法分别采用FISH和IHC检测414例乳腺癌组织Her-2基因扩增的情况,对两种方法检测结果进行统计学比较分析。结果两种方法检测结果有统计学差异(P=0.000),与FISH的一致性在IHC检测c-erbB-2(+++)和(-～+)组较好Kappa值=0.643,两者一致率为83.2%,而在c-erbB-2(++)组一致性较差。结论 IHC可以作为初步筛查乳腺癌Her-2基因状态的首选方法。在c-erbB-2(+++)和(+～-)中常可以用IHC代替FISH检测HER-2基因扩增状况,而对于c-erbB-2(++)则应常规进行FISH检测。     

荧光原位杂交检测脑膜瘤中HER2基因状态

目的 探讨脑膜瘤中HER2基因扩增与HER2蛋白过表达的关系.方法 运用免疫组织化学(IHC)和荧光原位杂交(FISH)技术,对手术切除的80例脑膜瘤石蜡标本进行HER2状态的检测.结果 80例中,26例HER2蛋白(+)(32.50%),其中HER2基因扩增7例(8.75%);10例HER2蛋白(+++/++)(12.50%),HER2基因扩增7例(8.75%),占高表达组的100.00%;70例HER2蛋白(+/0)(87.50%)中,无HER2基因扩增.结果 HER2蛋白(+++/++)与HER2基因扩增密切相关.     

IHC和FISH联合检测278例浸润性乳腺癌中HER-2状态的研究

目的：同时应用免疫组织化学（IHC）和荧光原位杂交（FISH）技术,检测278例浸润性乳腺癌组织中的人类表皮生长因子受体2（Human Epidermal Growth Factor Receptor2,HER-2）,以探讨两种方法检测HER-2状态的符合率。方法：用IHC和FISH法分别检测278例浸润性乳腺癌组织中HER-2蛋白的表达和HER-2基因状态,以比较两种方法的阳性率和准确率。结果：IHC和FISH检测HER-2的总符合率为83.9%,Kappa系数为0.650（P〈0.001）。130例IHC标记HER-2结果显示为＋＋的病例中,44例（33.8%）Fish为阳性,54例结果1HC为阳性或＋的病例中的5例（9.3%）,经FISH检测证实HER-2基因有扩增。而94例IHC结果显示为＋＋＋的病例中,有3例（20.2%）FISH结果为阴性。在130例IHC显示＋＋的病例中,3例FISH结果为模棱两可。结论：IHC和FISH检测HER-2的符合率为83.9%,对IHC标记HER-2结果为＋＋的病例,用FISH检测大多能得到满意结果。对极少数HER-2基因扩增为临界状态的乳腺癌病例需结合其他检查结果综合考虑。     

胃癌HER2蛋白表达及基因扩增分析

目的 探讨胃癌组织HER2蛋白表达和基因扩增情况及其临床意义.方法 收集2011年4月至2012年5月间青岛大学医学院附属医院黄岛院区收治的80例胃癌标本,采用免疫组织化学染色（IHC）和显色原位杂交（CISH） 方法检测HER2蛋白表达和基因扩增情况,分析其与临床病理因素的相关性.结果 80例患者中,HER2蛋白表达（-）51例,（＋）12例,（＋＋）12例,（＋＋＋）5例,阳性表达率为21.3％（17/80）;HER2基因扩增 4例,临界扩增3例,基因扩增率为8.8％（7/80）.IHC和CISH检测结果呈正相关（P＜0.05）,两者的符合率为85.0％（68 /80）.HER2表达与肿瘤部位、分化程度及TNM分期有关（均P＜0.05）.结论 胃癌组织中有较高的HER2蛋白表达和基因扩增率,其与胃癌的发生发展密切相关.     

C-erbB-2基因与HER2蛋白用于浅表性膀胱癌复发的研究

目的探讨C-erbB-2基因扩增和HER2蛋白阳性表达与浅表性膀胱癌的术后复发和肿瘤分级、临床分期的关系。方法用荧光原位杂交技术（FISH）和免疫组化（IHC）法,分析经病理证实、随访3年以上的20例复发性膀胱肿瘤与20例未复发膀胱肿瘤的C-erbB-2基因扩增与HER2蛋白表达情况。结果在复发的膀胱癌组中,HER2蛋白表达阳性的共有7例;未复发组中1例HER2蛋白表达阳性,复发组明显高于未复发组;结论HER2蛋白的阳性表达与膀胱癌的复发具有相关性,而与肿瘤的分级、分期无统计学意义。C-erbB-2基因与膀胱癌的复发、分级、分期以及与HER2蛋白的过表达无相关关系。     

HER2低表达乳腺癌的临床病理学特征及其新辅助化疗疗效的相关影响因素分析

目的 探讨人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)阴性乳腺癌中不同程度HER2蛋白表达者的临床病理学特征及HER2低表达乳腺癌新辅助化疗疗效的相关影响因素。方法 回顾性收集161例西南医科大学附属医院乳腺外科2019年3月至2021年3月期间初治行新辅助化疗的HER2阴性乳腺癌患者的临床病理资料,分析不同程度HER2蛋白表达者临床病理学特征的差异,运用非条件logistic回归模型分析HER2低表达乳腺癌患者新辅助化疗后病理完全缓解（pathologic complete response,pCR）率的影响因素。结果 161例HER2阴性乳腺癌患者中HER2低表达者共108例,占67.1%;HER2低表达患者比HER2零表达[免疫组织化学染色（immunohistochemistry,IHC） 0]患者有更高的腋窝淋巴结转移率（P=0.048）,更低的组织学分级（P=0.006）,激素受体表达阳性者占比更高（P<0.001）;pCR率在HER2 IHC 0、IHC 1+和IHC2+/原位杂交（in sit...     

乳腺癌中HER-2基因的扩增及意义

目的 研究乳腺浸润性导管癌HER-2基因的扩增情况及其与乳腺癌临床病理因素的关系。方法 采用荧光原位杂交法（FISH）检测254例乳腺浸润性导管癌组织中HER-2基因的扩增情况,分析其与乳腺癌的肿瘤大小、组织学分级、腋淋巴结转移情况、病理分期、雌、孕激素受体表达以及该基因蛋白表达等的关系。结果 FISH检测到254例乳腺癌中有175例发生HER-2基因高扩增,该基因高扩增与乳腺癌的肿瘤大小、组织学分级、病理分期、腋淋巴结转移数、雌孕激素受体表达、HER-2蛋白表达均有关（P均＜0.05）,与患者的年龄无关 (P＞0.05)。结论 乳腺癌中HER-2基因的高扩增导致其蛋白过度表达,HER-2基因高扩增与乳腺癌的生长、恶性程度有关,是判断乳腺癌生物学行为的重要指标。联合检测HER-2基因扩增情况及其他指标有助于指导临床判断乳腺癌预后并制定治疗方案。     

乳腺癌Her-2与TOP2A基因表达对新辅助化疗效果的预测价值

目的探讨乳腺癌HER 2及TOP2A基因的表达及其与蒽环类药物化疗效果的关系。方法收集我院4年间收治的新辅助化疗患者5 8例标本,采用FISH方法检测所有患者新辅助化疗前癌组织HER 2及TOP2A基因的表达情况;全组患者采用FEC方案行新辅助化疗。化疗4周期后按RECIST标准评估疗效;分析HER 2和TOP2A基因的扩增与疗效的关系。结果全组患者HER-2基因扩增19例(1 9/5 8,3 2.8%),TOP2A基因扩增1 1例(1 1/5 8,1 9.0%),HER 2与TOP2A共扩增1 1例(1 1/5 8,1 9.0%);HER-2基因、TOP2A基因及HER-2和TOP2A基因共扩增均与化疗效果呈正相关[rs=0.52(P0.0 5),rs=0.5 3(P0.0 5)及rs=0.5 6(P0.0 5)]。结论乳腺癌组织中HER 2及TOP2A的扩增均对蒽环类化疗药物的疗效有预测作用,两者的共扩增更有利于预测蒽环类药物的疗效。     

Human epidermal growth factor receptor type 2 protein expression in Chinese metastatic prostate cancer patients correlates with cancer specific survival and increases after exposure to hormonal therapy

Aim： To investigate human epidermal growth factor receptor type 2 （HER2） protein expression and gene amplification in Chinese metastatic prostate cancer patients and their potential value as prognostic factors. Methods： Immunohistochemistry （IHC） was performed to investigate HER2 protein expression in prostate biopsy specimens from 104 Chinese metastatic prostate cancer patients. After 3-11 months of hormonal therapy, 12 patients underwent transurethral resection of the prostate （TURP）. HER2 protein expression of TURP specimens was compared with that of the original biopsy specimens. Of these, 10 biopsy and 4 TURP specimens with HER2 IHC staining scores ≥ 2＋ were investigated for HER2 gene amplification status by fluorescent in situ hybridization （FISH）. Results： Of the 104 prostate biopsy specimens, HER2 protein expression was 0, 1＋, 2＋ and 3＋ in 49 （47.1%）, 45 （43.3%）, 8 （7.7%） and 2 （1.9%） cases, respectively. There was a significant association between HER2 expression and Gleason score （P = 0.026）. HER2 protein expression of prostate cancer tissues increased in 33.3% of patients after hormonal therapy. None of the 14 specimens with HER2 IHC scores 〉 2＋ showed HER2 gene amplification. Patients with HER2 scores 〉 2＋ had a significantly higher chance of dying from prostate cancer than those with HER2 scores of 0 （P = 0.004） and 1＋ （P = 0.034）. Multivariate Cox regression analysis showed that HER2 protein expression intensity was an independent predictor of cancer-related death （P = 0.039）. Conclusion： An HER2 IHC score 〉 2＋ should be defined as HER2 protein overexpression in prostate cancer. Overexpression of HER2 protein in cancer tissue might suggest an increased risk of dying from prostate cancer. HER2 protein expression increases in some individual patients after hormonal therapy.     

The influence of polysomy 17 on HER2 gene and protein expression in adenocarcinoma of the breast: a fluorescent in situ hybridization, immunohistochemical, and isotopic mRNA in situ hybridization study

Breast carcinomas with amplification of HER2 on chromosome 17 are associated with HER2 protein overexpression, adversely affecting prognosis and predicting response to Herceptin therapy. Chromosome 17 polysomy is encountered in assessing HER2 gene status, and its impact on HER2 gene and protein expression remains unclear. This impact was investigated in breast carcinomas identified by fluorescence in situ hybridization (FISH) to have a gain of chromosome 17 (CEP17+; n = 56), using a dual probe assay, which detects HER2 gene copy number and enumerates chromosome 17 (HER2/CEP17; Vysis). Cases were immunostained for HER2 protein (CB-11, Ventana), and scored blinded to FISH. A subgroup was evaluated by isotopic in situ hybridization for HER2 mRNA expression. Controls included ten HER2 amplified and ten nonamplified tumors, eusomic for chromosome 17. Immunohistochemistry (IHC) for HER2 protein was negative (0 or 1+) in 69% (39 of 56), 2+ in 27% (15 of 56), and 3+ in 3% (2 of 56) of CEP17+ cases. The mean CEP17 copy number among the three groups was similar (3.1, 3.0, and 3.1 for IHC 0/1+, 2+, and 3+, respectively). Isotopic in situ hybridization for HER2 mRNA performed on 26 CEP17+ cases (16 IHC 0-1+, 10 IHC 2+ or 3+) showed no increased HER2 mRNA expression (normalized to beta-actin mRNA). The mRNA expression and the IHC staining of the HER2-amplified and nonamplified controls was concordant with their FISH status. These results suggest that chromosome 17 polysomy in the absence of HER2 amplification does not have a significant biologic influence on HER2 gene expression in breast carcinoma.     

HER-2/neu Gene Amplification in Esophageal Adenocarcinoma and Its Influence on Survival

Background

HER-2/neu (c-erbB-2, HER2) gene amplification and protein overexpression have been associated with poor prognosis in several solid tumors, including breast and gastric cancer. Its incidence and significance in esophageal adenocarcinoma is unknown.

Materials and Methods

Tissue microarrays were successfully constructed from 89 paraffin-embedded archival specimens of esophageal adenocarcinomas for HER2 gene amplification by silver-enhanced in situ hybridization (SISH). No patients had undergone neoadjuvant therapy. Protein overexpression was tested with immunohistochemistry (IHC) using automated immunostaining (Ventana Benchmark). Incidence of HER2 positivity, correlation to clinicopathological variables in esophageal cancer patients, and concordance between SISH and IHC were determined.

Results

True HER2 gene amplification was detected in 14 esophageal cancer specimens (16%), and 92% of those with high-level HER2 amplification showed positive HER2 protein overexpression. No significant associations were found among gene amplification and clinicopathological factors. The 5-year survival rates were 57% for esophageal cancer patients with HER2 amplification compared with 32% without, but the difference in overall survival was not significant (P = .37). The correlation between SISH and IHC was statistically significant (P < .0001).

Conclusion

While molecular targeting may be possible for approximately 16% of esophageal adenocarcinoma patients, HER2 oncogene amplification did not influence survival in this study.

     

Her-2/neu gene amplification in low to moderately expressing breast cancers: possible role of chromosome 17/Her-2/neu polysomy

Overexpression of the Her-2/neu (HER2) oncogene is known to confer important prognostic and predictive value to patients with breast cancer. Controversy exists as to the best method for its determination caused primarily by the variable sensitivities of the different antibodies and interobserver differences, particularly in the group of breast cancers with borderline levels of expression of the protein product. This study was therefore designed to determine the status of the HER2 gene amplification in a group of breast carcinomas with low levels of overexpression. After an initial validation of our procedures, a series of 52 consecutive cases of formalin-fixed, paraffin-embedded breast cancers with low levels of overexpression and a series of 22 cases with no expression by immunohistochemistry were analyzed by fluorescence in situ hybridization (FISH), and the results correlated statistically. Amplification of the HER2 gene was observed in 16% of equivocal to weakly positive cases. Those that were amplified showed low levels of amplification with ratios less than 4.5 and a characteristic scattered pattern of distribution of HER2 signals in the FISH assay. In addition, heterogeneity was noted in two cases in the amplification of the HER2 gene within the same tumor samples with pockets of amplified tumor cells amidst nonamplified tumor cells. In cases without amplification, a statistically significant number showed chromosome 17 polysomy. In conclusion, equivocal to low levels of HER2 overexpression in breast cancers are associated, in the majority of cases, with chromosome 17 polysomy and a corresponding increase in the HER2 gene numbers. True gene amplification is present in only a minority of cases. FISH analysis should be used for confirmation of gene amplification. Prior screening and selection of appropriate immunohistochemistry-positive areas for FISH analysis may prove beneficial.     

Predicting the HER2 status of breast cancer from basic histopathology data: an analysis of 1500 breast cancers as part of the HER2000 International Study

The tests that are currently available for the measurement of overexpression of the human epidermal growth factor-2 (HER2) in breast cancer have shown considerable problems in accuracy and interlaboratory reproducibility. Although these problems are partly alleviated by the use of validated, standardised 'kits', there may be considerable cost involved in their use. Prior to testing it may therefore be an advantage to be able to predict from basic pathology data whether a cancer is likely to overexpress HER2. In this study, we have correlated pathology features of cancers with the frequency of HER2 overexpression assessed by immunohistochemistry (IHC) using HercepTest (Dako). In addition, fluorescence in situ hybridisation (FISH) has been used to re-test the equivocal cancers and interobserver variation in assessing HER2 overexpression has been examined by a slide circulation scheme. Of the 1536 cancers, 1144 (74.5%) did not overexpress HER2. Unequivocal overexpression (3+ by IHC) was seen in 186 cancers (12%) and an equivocal result (2+ by IHC ) was seen in 206 cancers (13%). Of the 156 IHC 3+ cancers for which complete data was available, 149 (95.5%) were ductal NST and 152 (97%) were histological grade 2 or 3. Only 1 of 124 infiltrating lobular carcinomas (0.8%) showed HER2 overexpression. None of the 49 'special types' of carcinoma showed HER2 overexpression. Re-testing by FISH of a proportion of the IHC 2+ cancers showed that only 25 (23%) of those assessable exhibited HER2 gene amplification, but 46 of the 47 IHC 3+ cancers (98%) were confirmed as showing gene amplification. Circulating slides for the assessment of HER2 score showed a moderate level of agreement between pathologists (kappa 0.4). As a result of this study we would advocate consideration of a triage approach to HER2 testing. Infiltrating lobular and special types of carcinoma may not need to be routinely tested at presentation nor may grade 1 NST carcinomas in which only 1.4% have been shown to overexpress HER2. Testing of these carcinomas may be performed when HER2 status is required to assist in therapeutic or other clinical/prognostic decision-making. The highest yield of HER2 overexpressing carcinomas is seen in the grade 3 NST subgroup in which 24% are positive by IHC.     

Detection of HER-2/neu gene amplification in breast cancer using a novel polymerase chain reaction/ligase detection reaction technique

BACKGROUND: Gene amplification is the primary mechanism of HER-2/neu overexpression in breast cancer and is a strong predictor of prognosis. Currently screening for HER-2/neu gene amplification in breast cancer is done by fluorescent in-situ hybridization (FISH), which is accurate but costly and labor intensive. We have evaluated a new PCR (polymerase chain reaction)-based assay for the detection of HER-2/neu gene amplification in human breast cancer. STUDY DESIGN: A total of 15 breast cancer cell lines and 14 breast cancer specimens were evaluated. HER-2/neu status of the tumors was evaluated by FISH and then assessed using a quantitative polymerase chain reaction/ligase detection reaction (PCR/LRD) technique. RESULTS: Amplification of the HER-2/neu gene was detected in seven cell lines previously reported to have amplification and no amplification was found in any of the six that had been reported not to have amplification. In the assessment of breast specimens the PCR/LDR and FISH assays were in complete agreement. All 10 tumors with amplification by FISH were also amplified by PCR/LDR. CONCLUSIONS: The PCR/LDR technique successfully detects HER-2/neu gene amplification in clinical breast cancer specimens and shows 100% concordance with FISH. This technique is an accurate and rapid alternative to FISH with the potential for automation and high throughput analysis of HER-2/neu status in breast cancer.