荧光原位杂交检测乳腺癌HER2基因状态

目的 探讨荧光原位杂交（FISH）,与免疫组织化学（IHC）染色（3＋）和（2＋）检测HER2基因状态的结果的一致性、导致两者差异的可能原因及FISH检测HER2基因状态的必要性和可行性。方法 采用PathVysion^TM探针试剂盒,以FISH方法,对28例IHC EnVision法染色分别为（3＋）、（2＋）、（1＋）和阴性（-）的乳腺癌石蜡切片标本进行HER2基因状态的检测。结果 HER2表达（3＋）的12例标本中,10例HER2基L大1扩增,其中2例为17号染色体多体,另2例无扩增的病例均为多体;IHC为（2＋）的10例标本中,7例为HER2基因扩增,其中1例为多体,另3例无扩增病例中2例为多体;IHC为（1＋）的3例标本均无HER2基因扩增,其中1例为多体;IHC（-）的3例标本均无基因扩增,其中1例为多体。结论 IHC是初步筛查HER2状态的首选方法;因IHC（2＋）与FISH检测结果差异较大,所以这类患者应做FISH确诊;IHC（3＋）存在假阳性,主要原因可能是由于17号染色体非整倍体,必要时这类患者也应做FISH。     

显色原位杂交和免疫组织化学检测乳腺癌HER2/neu基因状况和蛋白表达的对照性研究

目的探讨显色原位杂交（CISH）在检测乳腺癌中HER2／neu基因扩增上的作用。方法挑选乳腺浸润性导管癌患者组织石蜡蜡块（回顾性255例,前瞻性271例）,进行免疫组织化学（IHC）、CISH检测。15例回顾性标本送往德国HERA检测中心进行FISH检测。结果（1）回顾性病例中IHC阳性3＋者CISH基因扩增率为91．6％（120／131）,IHC2＋者CISH基因扩增率为56．5％（39／69）,IHC与CISH检测结果符合率为81．2％（207／255）,两者明显相关（P〈0．01）。（2）前瞻性病例中IHC蛋白过表达率31．7％．CISH基因扩增率27．3％。IHC3＋者CISH基因扩增率为91．4％（53／58）,IHC2＋者CISH基因扩增率为46．4％（13／28）,IHC与CISH检测结果符合率为89．7％（243／271）,两者明显相关（P〈0．01）。（3）经德国检测中心荧光原位杂交（FISH）检测的15例中14例和CISH结果完全一致,1例检测失败,而CISH为无扩增。（4）CISH检测基因扩增与雌激素受体（ER）、孕激素受体（PR）表达明显负相关（P值均〈0．01）。结论CISH检测HER2基因扩增结果与IHC检测蛋白过表达及FISH结果高度一致,CISH是检测HER2基因扩增的一项新技术。     

结直肠癌EGFR和HER2蛋白表达及其基因拷贝数分析

目的探索结直肠癌EGFR和HER2蛋白表达和基因拷贝数增加的频率,以及蛋白表达与基因拷贝数增加之间的相关性。方法以石蜡包埋组织芯片为材料,分别采用免疫组织化学（EGFR pharmDx^TM和HercepTest^TM试剂盒）和荧光原位杂交（FISH,LSI EGFR SpectrumOrange／CEP7 SpectrumGreen探针组合和Path V ysion^TM试剂盒）方法,检测42例中国人结直肠癌EGFR和HER2蛋白表达和基因状态。结果EGFR免疫组织化学0者18例、1＋者10例、2＋者5例、3＋者9例;HER2免疫组织化学0者39例、1＋者1例、2＋者1例、3＋者1例。通过FISH分析,EGFR基因拷贝数无明显增高18例（42．9％）,其中包括二体性14例（33．3％）、低三体性4例（9．5％）;EGFR基因拷贝数相对增高者24例（57．1％）,包括高三体性3例（7．1％）、低多体性9例（21．4％）、高多体性12例（28．6％）;本组患者中无EGFR基因扩增者。HER2基因拷贝数增加者共4例（9．5％）,其中包括基因扩增1例（2．4％）。EGFR的蛋白表达与基因拷贝数增加无相关性,两指标与肿瘤分化之间也无相关性。HER2免疫组织化学3＋和基因扩增结果一致,与乳腺癌相似。结论这组患者中,具有较高的EGFR蛋白表达和基因拷贝数增加的比率,但蛋白表达与基因状态之间以及二者与肿瘤分化程度之间都没有相关性;HER2表达和基因拷贝数增加的频率较低。     

1170例女性乳腺癌HER2基因状态的荧光原位杂交结果分析

目的 以大样本量准确地检测乳腺浸润性导管癌的HER2基因各种异常类型及其频率.方法 采用PathVysionTM探针试剂盒,以荧光原位杂交(FISH)方法,分析1170例拟采用曲妥珠单抗治疗或相关化疗的女性乳腺癌患者石蜡切片标本的HER2基因拷贝状态.结果 直接用FISH分析的1170例标本中,408例(34.87%)阴性,762例(65.13%)阳性,其中信号成簇扩增者87例,占全部阳性病例的11.42%;其余675例中,低度扩增159例(23.56%),中度扩增422例(62.52%),高度扩增94例(13.93%).1170例中,FISH结果为临界值(结果不确定)的14例(1.20%),其中比值为1.8-2.0的HER2阴性临界者1.23%(5/408),比值为2.0-2.2的HER2阳性临界者1.18%(9/762);17号染色体非整体性发生率为73.00%(854/1170),其中亚二体性为22.65%(265/1170),低多体性为38.38%(449/1170),高多体性为11.97%(140/1170);17号染色体多体性50.34%.阳性762例中,二体性182例(23.88%),亚二体性184例(24.15%),低多体性300例(393.37%),高多体性96例(12.60%);17号染色体多体性51.97%.阴性408例中,二体性134例(32.84%),亚二体性81例(19.85%),低多体性149例(36.52%),高多体性44例(10.78%).17号染色体多体性47.30%.HER2基因单等位基因缺失者占1170例之2.39%,17号染色体单体性在FISH阳性病例中占5.00%(38/762),在FISH阴性病例中占4.41%(18/408);HER2比值<1.5者占1170例之32.30%,占FISH阴性408例之92.65%;比值介于1.5-2.2之间的占9.23%(108/1170).结论 经临床筛选出拟进行生物靶向及相关治疗的中国内地女性乳腺癌患者FISH检测HER2扩增率高,其中以HER2基因中度扩增为常见类型;17号染色体非整体性,尤其是17号染色体多体性十分常见,其意义值得关注.     

显色原位杂交检测乳腺癌HER2基因扩增的应用价值

目的评估显色原位杂交（CISH）技术在检测乳腺癌标本中HER2基因扩增的应用价值。方法分别采用免疫组织化学（IHC）EnVision和CISH两种方法,检测165例乳腺癌中HER2蛋白表达及HER2基因扩增的情况。结果（1）IHC检测HER2蛋白表达阴性者107例,表达阳性1＋者24例,CISH均未检测到HER2基因扩增;（2）IHC检测HER2表达3＋者22例,CISH检测中21例呈现HER2基因的高倍扩增,仅1例呈低倍扩增,HER2基因高倍扩增及其蛋白表达之间的符合率为95．5％;（3）IHC检测HER2表达2＋者12例,CISH检测有3例HER2基因呈高倍扩增,6例呈低倍扩增,3例无扩增。结论CISH在检测HER2基因扩增与IHC检测的结果具有较高的一致性和敏感性,可以作为一种检测乳腺癌HER2基因状况的方法。     

HER2免疫组织化学1＋浸润性乳腺癌的基因扩增状态和mRNA表达分析北大核心CSCD

目的探讨HER2免疫组织化学（IHC）1＋乳腺癌患者的基因扩增状态和原位HER2 mRNA表达情况,为精准检测HER2及相关临床策略的修订提供初步参考数据。 方法抽取北京协和医院2011至2013年间手术获取的非特殊型浸润性乳腺癌HER2 IHC 1＋存档蜡块标本65例,制作组织芯片,分别采用荧光原位杂交（FISH）和RNAscope方法检测其HER2基因状态和原位mRNA表达水平。如原发灶HER2的FISH结果不确定或阴性但mRNA高表达,再以FISH复检其淋巴结转移灶的HER2基因状态。结果在65例标本中,FISH阳性4例（6.2%）,其中2例为HER2/CEP17〉2且平均HER2基因拷贝数〉4,另2例为HER2/CEP17〈2但平均HER2基因拷贝数〉6;在此4例FISH阳性标本中,2例为mRNA高表达（RNAscope 3分）,另2例为中度表达（RNAscope 2分）。另外,在全部标本中,3例HER2/CEP17〉2但平均HER2基因拷贝数〈4,其中2例mRNA高表达（RNAscope 3分和4分）,另1例mRNA中度表达（RNAscope 2分）。未见HER2基因状态及mRNA表达与临床病理特征（肿瘤大小、分化程度、淋巴结转移、脉管癌栓）之间存在明显相关性（P均〉0.05）。结论在本组HER2 IHC 1＋乳腺癌中检出了部分FISH阳性的标本,其mRNA也有一定程度的表达,可判断为抗HER2治疗的潜在获益者。鉴于精准分子分型对于乳腺癌临床处理及相关诊疗策略制定具有重要意义,后续有必要进行大样本量的验证。     

肝细胞癌中p53基因的缺失与HER-2基因的扩增及其意义

目的　检测原发性肝细胞癌中 17号染色体上p5 3基因的缺失与HER 2基因的扩增并探讨其临床意义。方法　分别以不同颜色荧光标记的p5 3基因、HER 2基因和 17号染色体着丝粒为双色探针组 ,应用间期双色荧光原位杂交 (FISH)技术检测 42例肝细胞癌间期细胞核中p5 3基因、HER 2基因和 17号染色体的拷贝数及其相对比例 ,以确定其缺失或扩增 ,结合临床分期、血清甲胎蛋白 (AFP)、肿瘤大小等资料进行统计分析其临床意义。结果　 42例肝细胞癌中p5 3基因缺失 2 7例( 64 3 % ) ;HER 2基因扩增 9例 ( 2 1 4% ) ,其中高拷贝扩增 (HC) 4例 ( 9 5 % ) ,低拷贝扩增 (LC) 5例( 11 9% ) ,同时具有缺失与扩增的 6例 ( 14 3 % ) ;另外 ,17号染色体多倍体有 2 6例 ( 61 9% )。p5 3基因缺失与 17号染色体多倍性呈正相关 ( χ2 =12 2 86,P <0 0 0 1) ,但与HER 2基因的扩增无关 ( χ2 =0 0 0 ,P =1 0 0 )。p5 3基因缺失与肝细胞癌患者AFP水平、肿瘤大小有关 (P <0 0 5 ) ;术后 2年存活率p5 3基因缺失患者 ( 18 5 % )显著低于无缺失患者 ( 60 0 % ,χ2 =7 467,P =0 0 0 6)。结论　原发性肝细胞癌的17号染色体上存在p5 3基因高频缺失和HER 2基因的扩增 ,它们可能与部分原发性肝细胞癌的发生发展有关     

乳腺癌患者17号染色体表达状态及临床病理意义探讨

目的观察乳腺癌患者17号染色体倍体性及临床病理意义。方法应用荧光原位杂交(FISH),技术检测乳腺癌石蜡切片标本的HER2基因状态,17号染色倍体性与临床病理的关系。结果 56例患者17号染色体倍体异常占35.71%(20/56),HER2基因扩增和不扩增病例17号染色体倍体异常分别为50%(8/16)和30%(12/40),二者比较有显著差异(χ2=16.73,P0.01);多倍体主要分布在基因扩增病例中(75%,6/8),亚二倍体主要为基因不扩增病例(91.67%,11/12);多倍体患者发病年龄大,绝经发生率高,生产次数多,ER和PR表达率低,与二倍体和亚二倍体组比较有显著或极为显著性差异(P0.05或P0.01),二倍体和亚二倍体组比较,差异不显著(P0.05)。结论 17号染色倍体异常是乳腺癌患者常见的遗传学改变,可能与HER2基因表达状态有关;多倍体患者具有独特的生物学特征,与更多的临床预后不良因素有关。     

前列腺癌8号染色体改变与Gleason评分之间的相关性

目的探索前列腺癌8号染色体数目增加及c-myc基因和脂蛋白脂酶（LPL）基因状态和发生频率,分析8号染色体改变与前列腺癌Gleason评分之间的相关性及其在前列腺癌发生发展中的作用。方法采用ProVysion^TM三色探针组合,以荧光原位杂交（FISH）方法检测34例未经临床治疗的前列腺癌穿刺组织石蜡切片标本的8号染色体改变,其中包括Gleason评分5分者1例,6分者10例,7分者14例,8分者4例,9分者5例,并进行8号染色体各种异常之间及其与前列腺癌Gleason评分级别之间的关联性分析。结果8号染色体增加为17／34（50％）,c-myc基因拷贝数增加为21／34（61．8％）,LPL单体为15／34（44．1％）,c-myc基因扩增为23／34（67．6％）,LPL基因缺失为21／34（61．8％）,同时具有LPL基因缺失和c-myc基因扩增为16／34（47．1％）,至少有其中一种遗传学异常者为29／34（85．3％）。8号染色体增加与Gleason评分级别增高呈明显的正相关关系（P=0．0006）;c-myc基因拷贝数增加与Gleason评分级别增高呈正相关关系（P=0．0035）;LPL缺失与Gleason评分级别增高呈负相关关系（P=0．0383）;调整年龄后,除了上述三个变量与Gleason评分级别的相关关系仍然存在以外,c-myc基因扩增与Gleason评分级别增高也呈现正相关关系（P=0．0462）。结论8号染色体数目增加、c-myc基因拷贝数增加、c-myc基因扩增和LPL基因丢失都与Gleason评分级别有关,c-myc基因扩增同时伴有LPL基因缺失也是前列腺癌的遗传学特征之一,提示8号染色体异常可能与前列腺癌的发生和进展有关。     

非小细胞肺癌EGFR和HER2基因状态及相关性

目的探讨EGFR和HER2基因变化在非小细胞肺癌发生发展中的作用及其二者相关性。方法采用PathVysion^TM探针试剂盒和LSI EGFR SpectrumOrange／CEP7 SpectrumGreen探针组合,对31例中国人非小细胞肺癌石蜡切片标本,其中腺癌20例、鳞癌2例、大细胞癌2例、细支气管肺泡癌4例、腺鳞癌3例,分别进行EGFR和HER2基因状态的检测,并统计分析二者之间的相关性。结果EGFR基因扩增者6例;EGFR基因无扩增的25例中4例为四体,5例为多体,EGFR基因拷贝数增加和基因扩增者共15例（15／31）。HER2基因扩增者2例;HER2基因无扩增的29例中4例为三体,1例为四体,9例为多体,HER2基因拷贝数增加和基因扩增者共16例（16／31）。同时为EGFR和HER2基因异常（包括基因扩增和拷贝数增高）者12例（12／31）,而且几乎都是临床Ⅲ期至Ⅳ期的患者。EGFR和HER2基因异常之间的相关性X^2Adj=7．3045,P确切=0．0069。结论EGFR或HER2基因扩增在非小细胞肺癌中的发生率较低,但基因拷贝数增加是较常发生的事件。EGFR和HER2异常有相关性,EGFR和HER2基因都参与了非小细胞肺癌的发生发展,EGFR和HER2二聚体是肺癌常见的异源二聚体形式,而且它们的相互作用可能与疾病进展有关。     

HER2 status in sinonasal intestinal-type adenocarcinoma

Given that the prognosis of patients with sinonasal intestinal-type adenocarcinoma (ITAC) has not significantly changed recently, there is a desire for new therapeutic approaches to improve clinical management. HER2-targeted therapy has remarkably improved the overall survival of patients with HER2 amplified tumors. To date, HER2 assessment has produced contradictory results in ITAC. The aim of this study was to assess HER2 status at both protein and DNA levels in a large series of ITAC. HER2 status was assessed by immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) in forty-three patients that underwent surgical resection for ITAC at the Otorhinolaryngology Section, Padua University Hospital, between 2007 and 2016. IHC was evaluated using the four-tier score developed for gastroesophageal cancer. As for IHC, 83.7% (36/43) of ITAC were scored 0, 14% (6/43) 1+, and 2.3% (1/43) 2+. No HER2 amplification was detected by CISH. The present is the largest study of sinonasal ITAC tested with both IHC and CISH confirmation for HER2 status. No HER2 overexpression/amplification was detected. Contrary to previous studies, our findings seem to rule out any oncogenetic role of HER2 in ITAC pathogenesis.     

Single nucleotide polymorphisms of HER2 related to osteosarcoma susceptibility

Aims: The purpose of this study was to investigate the correlation between single necleotide polymorphisms (SNPs) of human epidermal growth factor receptor-2 (HER2) gene with osteosarcoma susceptibility in Chinese Han population. Methods: 90 patients with osteosarcoma and 100 healthy controls who were frequency-matched with the former by age and gender were enrolled for a case-control study. 5 SNPs of HER2, namely rs2952155, rs1810132, rs2952156, rs1136201 and rs1058808, were tested by Sequenom time of flight mass spectrometry technique. The linkage disequilibrium and haplotype were analyzed using haploview software. The risk intensity of osteosarcoma was expressed by odds ratio (OR) with 95% confidence interval (CI) which was calculated by chi-squared text. Hardy-Weinberg equilibrium (HWE) was also evaluated by chi-squared text. Results: HER2 gene rs1136201 and rs1058808 polymorphisms were associated with the increased risk of osteosarcoma (P=0.04 and 0.02, respectively). Allele G in rs1136201 was 1.67 higher risk for osteosarcoma in cases than the control group (OR=1.67, 95% CI=1.11-2.51) and G allele of rs1058808 polymorphism also significantly increased osteosarcoma susceptibility (OR=2.06, 95% CI=1.27-3.22). The haplotype analysis showed that haplotype C-T-G-G might be a susceptible haplotype to osteosarcoma (OR=1.74, 95% CI=1.01-3.00). HWE test was eligible in controls (P>0.05). Conclusion: HER2 gene rs1136201 and rs1058808 polymorphisms and haplotype C-T-G-G may be related to osteosarcoma susceptibility in Chinese Han population, indicating that the interaction of gene polrmorphism plays an role in osteosarcoma risk.     

Weekly Paclitaxel and Trastuzumab as a First-Line Therapy in Patients with HER2-Overexpressing Metastatic Breast Cancer: Magnitude of HER2/neu Amplification as a Predictive Factor for Efficacy

We evaluated the efficacy and safety of weekly paclitaxel plus trastuzumab as firs-tline chemotherapy in women with HER2-overexpressing metastatic breast cancer (MBC), and we investigated the prognostic factors including magnitude of HER2/neu amplification in this population. We analyzed 54 patients with HER2-overexpressing MBC that were treated with weekly paclitaxel plus trastuzumab as first-line chemotherapy from February 2004 to December 2006. At a median follow-up of 28 months, median time to progression (TTP) was 16.6 months (95% CI, 9.4 to 23.7 months) and median overall survival was 25.6 months (95% CI, 21.8 to 27.3 months). Therapy was generally well tolerated, although three patients (5.5%) experienced reversible, symptomatic heart failure. Of the 27 patients evaluable for the HER2 FISH, patients with a HER2/CEP17 ratio of ≤4.0 had significantly shorter TTP than those with a HER2/CEP17 ratio of >4.0 (10.8 vs. 23.2 months, P=0.034). A HER2/CEP17 ratio of >4.0 was identified as significant predictive factor of TTP by multivariate analysis (P=0.032). The combination of weekly paclitaxel plus trastuzumab as first-line chemotherapy is an effective regimen in patients with HER2-FISH-positive MBC. Furthermore, the magnitude of HER2 amplification is an independent predictive factor of TTP.     

HER3 and HER4 are highly expressed in human meningiomas

HER3 and HER4 are tyrosine kinase receptors of the ErbB family that have been detected in several cancers but lack substantial investigation in human meningiomas. In this study, HER3 and -4 expression levels were evaluated as potential biomarkers by immunohistochemistry and explored for association to clinical features in a large series of human meningiomas. 186 primary intracranial meningiomas from adult patients were investigated with antibodies against HER3 and -4 intracellular domains. Tumors were scored with a staining index (SI) based on cytoplasmic/membranous staining intensity and on the percentage of positive cells. SIs were tested for associations with WHO malignancy grade, tumor subtype, localization, and prognosis. HER3 and HER4 were highly expressed in most tumors. Both cytoplasmic and membranous immunoreactivity occurred, and for HER4 nuclear immunoreactivity was observed as well. Non-neoplastic meningeal tissue was not immunoreactive. HER3 and -4 immunoreactivity was not associated with WHO malignancy grade, nor with recurrence or survival in adjusted analyses. Meningiomas of all grades were shown to widely express both HER3 and HER4 receptors. This feature may have diagnostic value since non-neoplastic meninges were not immunoreactive. There was no prognostic significance in adjusted survival analyses.     

HER2-Testung beim Magenkarzinom

Valid HER2 testing is essential for the optimal care of patients with HER2-positive gastric cancer and the correct use of first-line treatment. Although all cases of breast cancer are routinely tested for the HER2 status, HER2 testing in gastric cancer has still not become part of the routine and is usually only done upon request by the therapist. An interdisciplinary group of German experts has taken on the challenges of HER2 testing in gastric cancer as an opportunity to address essential aspects and questions on the practical use of HER2 testing in this indication from the perspective of pathologists and therapists. The recommendations made in this article reflect the consensus of all participants and correspond to their opinions and long-term experience.