荧光原位杂交与免疫组化检测乳腺癌组织HER-2基因扩增或蛋白表达情况的比较分析

目的 评估免疫组织化学(IHC)法检测乳腺癌组织中HER-2蛋白表达和荧光原位杂交(FISH)法检测其HER-2基因扩增在乳腺癌分子靶向治疗中的临床意义.方法 采用IHC法及FISH法分别检测47例乳腺癌组织中HER-2蛋白表达及HER-2基因扩增情况.结果 47例乳腺癌石蜡标本中,HER-2蛋白表达(+++)20例(42.55%),(++)10例(21.28%),(+)17例(36.17%);HER-2基因扩增22例(46.81%),无扩增25例(53.19%),其中17号染色体多体5例(10.64%).两种检测结果呈正相关(rs=0.415,P<0.05).结论 IHC法检测乳腺癌组织中HER-2蛋白表达町以作为是否应用赫赛汀的初筛;HER-2蛋白表达阳性的病例有必要进一步进行HER-2基凶扩增检测,来确定临床治疗是否应用赫赛汀.     

荧光原位杂交法检测乳腺癌HER-2表达与腋淋巴结转移相关性的研究

目的:探讨用荧光原位杂交法(FISH)检测乳腺癌HER-2基因的表达与腋淋巴结转移的相关性.方法:采用免疫组化方法检测乳腺癌组织中HER-2的表达,再用FISH进一步检测阳性病例中HER-2基因的扩增情况,比较两者结果的差异,并且分析HER-2的表达与腋淋巴结转移的相关性.结果:免疫组化(IHC)法检测HER-2阳性42例,FISH检测阳性9例,FISH法检测HER-2基因与HER-2蛋白过表达之间相关(P＜0.05),并且IHC和FISH两种方法检测HER-2表达均与腋淋巴结转移有相关性(P＜0.05).结论:FISH可稳定检测HER-2基因的扩增,其表达与腋淋巴结转移相关.     

色素原位杂交法和免疫组织化学法检测乳腺癌HER-2基因扩增及蛋白表达

目的:对照色素原位杂交法(chromogenic in situ hybridization,CISH)和免疫组织化学法(immuno-histochemistry,IHC)检测乳腺癌组织中人表皮生长因子受体(human epidermal growth factor receptor-2,HER-2)基因扩增及其蛋白表达的状况.方法:采用CISH技术检测145例乳腺癌组织中HER-2的基因扩增情况,其中14例标本经过荧光原位杂交(fluorescence in si-tu hybridization,FISH)检测,随后分别用FISH和IHC方法检测的HER-2基因扩增及蛋白表达状况与之进行回顾性对照,并按照病理分级、淋巴结状态、绝经与否和雌激素受体(estrogen receptor,ER)/孕激素受体(progesterone receptor,PR)表达情况进行分层,分析HER-2表达与乳腺癌各高危因素之间的关系.结果:CISH检测发现,HER-2无扩增71例(50.0%),低扩增11例(7.6%),高扩增63例(43.4%).14例FISH与CISH检测结果比较,符合率为100.0%.145例IHC与CISH检测结果的符合率为84.8%(P<0.05).在IHC检测积分为0/ 以及 的病例中,HER-2基因扩增与蛋白表达情况基本一致,其符合率均在90%以上;而在IHC检测积分为 的标本中,HER-2基因扩增率仅为61.1%.CISH及IHC检测均显示ER/PR表达情况与HER-2阳性呈负相关,ER和PR均为阴性患者的HER-2基因扩增率和蛋白表达率明显高于ER/PR阳性的患者(CISH:68.3%vs 38.8%,P<0.01;IHC:71.7%vs 48.2%,P<0.01).HER-2状态与乳腺癌病理分级、腋淋巴结转移以及绝经与否无关(P>0.05).结论:CISH技术检测HER-2操作简便,准确性高,可以代替FISH技术,对IHC评分为 的病例应进一步确认HER-2状态.HER-2除了与ER/PR表达相关外,与其他乳腺癌高危因素无关,可作为独立指标进行检测.     

应用荧光原位杂交法检测乳腺癌石腊样本中HER-2 基因的扩增

 目的 探讨荧光原位杂交法（Fluorescence in situ hybridization，FISH）检测乳腺癌HER-2基因扩增在临床病理诊断及分子靶向治疗中应用的可能性。方法 用FISH技术和免疫组化（Immunohisto—chemistry，IHC）技术检测50例乳腺导管癌石蜡包埋标本并比较两种方法的结果以及与临床病理的关系。结果 16／50例HER-2蛋白表达阳性，其中强阳性5例，中度阳性9例，弱阳性2例；11／50（22％）例乳腺癌标本FISH技术检测HER-2基因扩增阳性，其中5／5为免疫组化HER-2蛋白强阳性病例；6／9为中度阳性病例，其中1例为17号染色体多倍体与HER-2基因扩增。HER-2基因扩增与蛋白表达与乳腺癌转移有关（P〈0．05）。结论 FISH技术可稳定地检测用IHC确定的HER-2蛋白阳性乳腺癌中HER-2基因的扩增状况，并用于临床赫赛汀分子靶向治疗病例的筛选。     

荧光原位杂交技术检测乳腺癌HER-2基因扩增实验方法的比较

目的:探讨荧光原位杂交(FISH)技术检测乳腺癌HER-2基因的最佳实验方法.方法:应用FISH技术检测55例乳腺癌组织标本的HER-2基因,分别采用水浴法和直接消化法,甲酰胺法和共变性法,检测HER-2基因表达成功率,对比实验方法的最佳结果.结果:两种消化法中,水浴法和直接法杂交成功率分别为86.21%(25/29)和88.46%(23/26),差异无统计学意义(P>0.05),杂信号比分别为17.24%(5/29)和46.15%(12/26),差异有统计学意义(P<0.05).两种变性法中甲酰胺法和共变性法杂交成功率分别为73.08%(19/26)和93.10%(27/29),差异有统计学意义(P<0.05).结论:FISH检测乳腺癌HER-2基因,以水浴法和共变性法杂交成功率最高.     

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。     

荧光原位杂交与免疫组织化学方法对乳腺癌Her-2检测的比较

[目的]探讨荧光原位杂交(FISH)与免疫组织化学方法(IHC)检测乳腺癌组织中人类表皮生长因子受体2(Her-2)表达结果的差异.[方法]采用FISH和IHC法分别检测508例乳腺癌标本中Her-2基因扩增和Her-2蛋白表达状况.[结果]在IHC法检测Her-2蛋白表达0/+的104例(20.47％)标本中,FISH检出13例(2.56％)Her-2基因扩增,89例(17.52％)为非扩增,2例(0.39％)为可疑;Her-2蛋白表达4++的245例(48.23％)标本中,FISH检出121例(23.82％)Her-2基因扩增,120例(23.62％)为非扩增,4例(0.79％)为可疑;IHC在Her-2蛋白表达+++的159例(31.30％)标本中,FISH检出137例(26.97％) Her-2基因扩增,22例(4.33％)为非扩增.[结论]对于IHC法初筛Her-2蛋白为++～+++的患者必须进行FISH检测,以明确Her-2基因的状态,从而指导临床医师诊断治疗.     

荧光原位杂交（FISH）检测乳腺癌中HER-2基因状态与p53、Ki-67、TOPOⅡ表达的相关性

目的:研究乳腺癌中HER-2基因状态和p53、Ki-67、TOPOⅡ蛋白表达的相互关系,以及HER-2基因扩增与HER-2蛋白表达的相关性。方法:运用FISH和IHC技术分别检测172例浸润性乳腺癌中HER-2基因状态及HER-2、p53、Ki-67、TOPOⅡ蛋白表达情况,分析HER-2基因状态与其相互的关系。结果:172例浸润性乳腺癌中HER-2基因扩增与p53、Ki-67、TOPOⅡ蛋白表达呈正相关（P<0.05）,HER-2基因扩增阳性率为43.6%（75/172）,HER-2基因扩增与HER-2蛋白表达具有相关性（P<0.05）。结论:联合检测HER-2基因状态和p53、Ki-67、TOPOⅡ蛋白表达可为浸润性乳腺癌的预后及分子靶向治疗提供依据。     

色素原位杂交法分析乳腺癌ＨＥＲ-２蛋白过表达者的基因状态

目的　采用色素原位杂交（ＣＩＳＨ）检测乳腺癌ＨＥＲ-２蛋白过表达者的ＨＥＲ-２基因状态,探讨ＨＥＲ-２蛋白表达与基因扩增的一致性。方法　采用Ｚｙｍｅｄ公司ＳｐｏＴ ＬｉｇｈｔＨＥＲ-２ＣＩＳＴ^ＴＭ试剂盒,按照美国临床肿瘤学会／美国病理医师学会（ＡＳＣＯ／ＣＡＰ）联合工作组推荐的结果判读标准,经免疫组织化学（ＩＨＣ）方法确认２３６例ＨＥＲ-２蛋白表达阳性,其中ＩＨＣ（＋＋）１４８例,ＩＨＣ（＋＋＋）８８例,对上述乳腺癌石蜡标本的ＨＥＲ-２基因行ＣＩＳＨ检测。结果　乳腺癌ＨＥＲ-２蛋白高表达者总基因扩增率为７０.３４％,其中ＨＥＲ-２表达ＩＨＣ（＋＋）者基因扩增率为５９.４６％（８８／１４８）,ＩＨＣ（＋＋＋）者基因扩增率为８８.６４％（７８／８８）,两者基因扩增状态差异有统计学意义（χ２ ＝２６.３５,Ｐ＜０.０１）。ＣＩＳＨ检测的ＨＥＲ-２基因扩增情况与雌激素受体（ＥＲ）、孕激素受体（ＰＲ）表达呈明显负相关（Ｐ＜０.０１）。结论　ＣＩＳＨ是一项简便经济的检测ＨＥＲ-２基因扩增技术,ＩＨＣ（＋＋＋）与ＣＩＳＨ阳性的一致性较高,但ＩＨＣ（＋＋）与ＣＩＳＨ阳性的符合率略低,有必要进一步行ＣＩＳＨ或荧光分子探针原位杂交（ＦＩＳＨ）检测明确基因扩增状态。     

乳腺癌组织微阵列HER-2和COX-2的表达及其相关性研究

目的:探讨HER-2和COX-2在乳腺癌组织中的表达及其意义。方法:构建乳腺癌组织微阵列,应用免疫组织化学SP法检测160例乳腺浸润性导管癌患者HER-2和COX-2的表达。结果:乳腺癌组织中HER-2和COX-2的阳性表达率分别为35%(56/160)和55·6%(89/160),且HER-2和COX-2的表达均与组织学分级、淋巴结转移、雌激素受体、孕激素受体及无病生存期显著相关,P<0·05或P<0·01;COX-2与HER-2的表达密切相关,P<0·01。结论:HER-2和COX-2的表达与乳腺癌的生物学行为有关,同时检测HER-2和COX-2可能对乳腺癌预后的判断及靶向治疗的选择提供帮助。     

非小细胞肺癌EGFR荧光原位杂交与免疫组化检测的比较

目的探讨荧光原位杂交（FISH）技术和免疫组化（IHC）法检测石蜡标本非小细胞肺癌（NSCLC）EGFR基因扩增及蛋白表达水平的差异性。方法采用FISH和IHC分别检测27例NSCLC患者石蜡标本EGFR基因和蛋白表达,对2种方法的检测结果进行对比分析。结果 14例IHC法EGFR表达（3＋）的标本中有9例FISH显示阳性（64.29%）,其中5例为EGFR基因高多体性扩增（55.56%）,4例为EGFR基因扩增（44.44%）;6例IHC（2＋）的标本中仅1例为高多体性扩增（16.67%）;2例IHC（1＋）及5例IHC（-）标本均无EGFR基因扩增。结论 IHC法初筛（3＋）、（2＋）的标本与FISH检测的符合率较低,提示对IHC检测EGFR表达为（3＋）及（2＋）并选择靶向药物治疗的病例,应采用FISH法对EGFR基因表达作进一步检测。     

Detection of Mycoplasma Hyorhinis Infection in Ovarian Cancer with in situ Hybridization and Immunohistochemistry

OBJECTIVE To detect Mycoplasma hyorhinis in ovarian cancer tissues and the relationship between mycoplasma infection and ovarian cancer. METHODS All specimens obtained from 109 cases with ovarian cancer were fixed in freshly prepared 10% neutral buffered formalin, embedded in paraffin, and cut into 4-μm sections for insitu hybridization (ISH) and then detected with immunohistochemistry (IHC). The expressions of 16S rRNA and P37 protein from mycoplasma hyorhinis were detected respectively using ISH and IHC. SPSS 13.0 so ware was employed to analyze the relationship between the results of the study and clinical pathological materials. RESULTS The expression rate of mycoplasma hyorhinis 16S rRNA gene and P37 protein was 20.2% (22/109) and 43.1% (47/109cases) in ovarian cancer tissues, respectively, but it was 0 (0/30cases) in the normal ovarian tissues. The difference in mycoplasma infection ratio between ovarian cancer tissues and normal tissues was extremely significant (P < 0.001). Anyhow, we didn't found any association between the mycoplasma infection and clinical pathological characters.CONCLUSION There was a mycoplasma infection in ovarian cancer tissues, which may play a role in oncogenesis of ovarian cancer.     

Practice of HER-2 Immunohistochemistry in Breast Carcinoma in Austria

Practice and accuracy of immunohistochemistry is known to vary highly. Reliability of HER-2 immunohistochemistry is critical because of its role in patient selection for therapeutical options in breast cancer. Therefore reliability of HER-2 immunohistochemistry in pathology laboratories in Austria was assessed. Ten tissue specimens of invasive ductal breast carcinomas and three cell line samples were tested. Presence/absence of gene amplification was determined by FISH to be used as a gold standard. Laboratories were asked to stain and assess slides using their routine immunohistochemical staining protocol. Overall the study consisted of 311 tests on tissue specimens and 142 on cell lines. In all cases manual scoring was performed. Participation was voluntary and was 94%. Overall sensitivity was 90.5% and specificity 99.2%. Overscoring including true false positive results were found in 6.7% and 6.3% in tissue specimens and cell lines, respectively. False negative determinations were obtained in 1.9% and 2.8% of tissue specimens and cell lines, respectively. HercepTest^TM showed slightly higher reliability in comparison with individualized staining methods. By manual scoring inaccurate scoring affected 12.3% of test results and 62% of the laboratories. In conclusion participation rate and accuracy of HER-immunohistochemistry was high all over the country. Manually performed scoring demonstrated some limitations.     

色素原位杂交在检测乳腺癌患者组织中HER2基因的应用

[目的]探讨色素原位杂交（CISH）在检测乳腺癌患者组织中HER2基因状态的临床应用，比较CISH与免疫组化（IHC）检测组织HER2状态的差异性。[方法]采用SPOT—Light HER2 CISH^TM试剂盒，以CISH方法对40例IHC EnVision法染色分别为（＋＋＋）、（＋＋）、（＋）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IHC（＋＋＋）的8例标本中，7例为HER2基因扩增CISH检测，1例无扩增；（＋＋）的13例标本中，5例为HER2基因扩增．8例无扩增；（＋）的10例标本中，2例为HER2基因扩增，7例无扩增；（-）的9例标本均无扩增。两种方法对乳腺癌组织HER-2／neu状态的检测有一定的差异性（kappa=0．458，P=0．003）。[结论]IHC是HER表达初步筛查的首选方法，由于蛋白表达和基因扩增检测结果存在一定的差异性，建议IHCC（＋＋＋～＋）患者进一步作CISH检测确诊。     

荧光原位杂交在膀胱尿路上皮癌中的应用

目的探讨3,7,17号染色体及9p21(p16基因)组合探针在监测膀胱尿路上皮癌术后复发及诊断膀胱尿路上皮癌的应用价值。方法利用荧光原位杂交（FISH）检测膀胱尿路上皮癌患者的尿脱落细胞及组织切片中3,7,17号染色体及9p21组合的畸变情况。结果FISH监测膀胱尿路上皮癌术后复发的敏感度为87.50%;术前FISH阳性的患者术后更易复发;FISH诊断膀胱尿路上皮癌的阳性率为78.85％。结论利用FISH检测尿脱落细胞中3,7,17号染色体及9p21组合的畸变能有效辅助监测膀胱尿路上皮癌术后复发;并可能在一定程度上预测术后复发;同时利用FISH检测组织切片中3,7,17号染色体及9p21组合的畸变也能有效辅助膀胱尿路上皮癌的诊断。     

Her2 Amplification Status in Iranian Breast Cancer Patients: Comparison of Immunohistochemistry (IHC) and Fluorescence in situ Hybridisation (FISH)

Introduction: Her2/neu is a biomarker which is amplified and/or overexpressed in a subset of breast cancerpatients who are eligible to receive trastuzumab. Her-2 gene amplification analysed by fluorescence in situhybridisation (FISH) and/or protein over-expression detected by immunohistochemistry (IHC) are the two mainmethods used to detect Her-2 status in clinical practice. The concordance rate between the two techniques iscontroversial. Methods: FISH analysis were performed on 104 tumoural samples from breast cancer patientswith known IHC results to determine the Her2 gene status. The FISH/IHC analyses results were then comparedand the concordance rate was determined. Results: Her2 gene amplification was detected in 0 of IHC score 1+,24/86 (27.91%) 2+, and 8/13 (61.54%) 3+. The IHC and FISH results concordance rates were 100%, 27.9%,and 61.5% for IHC scores of 1+, 2+, and 3+ respectively. Conclusion: The results of this study suggest that IHC1+ should be considered as negative while IHC 2+ results need further confirmative analysis by FISH. Furtherquality control and standardization of IHC technique are required to improve the concordance rate betweenthe two methods.     

膀胱癌荧光原位杂交检测及其临床意义

目的:分析膀胱移行细胞癌的染色体畸变情况,探讨荧光原位杂交(FISH)技术在膀胱癌的临床应用价值.方法:采用3、7、17号染色体着丝粒探针和9号染色体p16基因位点探针对56例膀胱移行细胞癌患者和20名健康人群的新鲜尿液进行FISH检测,统计染色体的畸变并分析其与病理分级、分期的关系.对所有病例同步进行尿细胞学分析.结果:膀胱癌患者尿脱落细胞核中3、7、17号染色体及9号染色体p16基因畸变率分别为58.9%、39.3%、58.9%和75.0%,各染色体畸变在膀胱癌不同分期之间的差异无统计学意义(P>0.05),3、7、17号染色体畸变在不同分级之间的差异具有统计学意义(P<0.05),四染色体探针组合诊断膀胱癌的总阳性率为80.4%;膀胱癌尿脱落细胞的FISH检出率明显高于尿细胞形态学.结论:膀胱癌的发生发展与染色体的畸变有关,膀胱癌尿脱落细胞的FISH检测,对膀胱癌的早期诊断、预后评估及复发监测等具有重要价值.