胃食管结合部癌Her－2基因扩增及蛋白表达与临床病理关系

目的：通过检测表皮生长因子－2（ Her－2）在胃食管结合部癌中的蛋白表达和基因扩增情况,探讨其与胃食管结合部癌患者的临床病理关系。方法采用免疫组织化学法（ IHC）和荧光原位杂交技术（ FISH）检测80例手术切除的胃食管结合部癌组织中Her－2的蛋白表达和基因扩增情况,并分析与胃食管结合部癌的病理分化程度、浸润深度、临床分期和淋巴结转移的关系。结果应用FISH技术检测显示在胃食管结合部癌组织中Her－2的蛋白阳性表达率为22．5％,应用IHC技术检测其在癌组织表达率为15％,Her－2基因扩增和蛋白阳性表达与胃食管结合部癌的分化程度和临床TNM分期有关（ P＜0．05）,而与年龄、性别和浸润深度及淋巴结转移无相关性。结论 Her－2蛋白过表达和基因扩增是胃食管结合部癌患者的一个预后指标,与其发生发展有关,联合检测Her－2蛋白表达水平及基因扩增程度对于胃食管结合部癌的治疗和预后评估有一定指导意义。     

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。     

肺癌的癌基因表达与临床的相关研究

myc基因家族(c-myc,N-myc,L-myc,c-myb)的扩增或过度表达与ras基因的点突变是肺癌的主要癌基因异常。c-myc基因的扩增或过度表达在人体多种肿瘤中出现,并与病人的不良预后有关。N-myc扩增和过度表达多出现在具有神经内分泌特性的肿瘤组织中,如神经母细胞瘤,小细胞肺癌等。N-myc     

HER2基因在胃癌组织中的表达及其临床意义

目的探讨HER2基因表达、扩增与胃癌生物学行为的关系。方法应用FISH技术,检测20例胃癌标本中HER2扩增情况,应用IHC技术检测胃癌组织中HER2表达情况。结果应用FISH技术检测胃癌组织中HER2基因扩增率为35%,IHC技术检测胃癌组织中HER2蛋白阳性表达率为30%;HER2基因扩增及蛋白阳性表达与胃癌浸润深度、淋巴结转移及病理分期有关(P<0.05)。结论 HER2基因表达与胃癌浸润和转移有关,可作为判断胃癌生物学行为和预后的重要指标。     

乳腺癌中HER-2基因及蛋白的检测和临床应用

目的:在蛋白及基因水平检测乳腺癌中HER-2的表达,比较免疫组化(IHC)及荧光原位杂交(FISH)法检测结果,分析其临床应用.方法: 应用免疫组化及荧光原位杂交法测定HER-2蛋白表达及基因扩增.结果: IHC检测HER-2蛋白,77例乳腺浸润性导管癌中 ,[(-)-(+)]占 35.06%,(++)占37.66%,(+++)占27.27%;FISH检测HER-2基因,扩增阳性率为51.95%.IHC强阳性组(+++),FISH阳性率为90.48%,IHC弱阳性组(++), FISH阳性率为 68.97%,IHC阴性组[(-)-(+)], FISH阳性率为 3.07%,P<0.01,具有显著统计学意义.以HER-2基因扩增为阳性标准,IHC 结果(+++)阳性预测值为90.48%,灵敏度为95.00%,特异度为92.86%;(++)阳性预测值为68.97%,灵敏度为95.24%,特异度为74.29%;[(-)-(+)]阴性预测值为96.30%.结论:HER-2蛋白过表达主要是基因扩增所致,IHC强阳性 (+++)及IHC阴性[(-)-(+)] 可直接指导临床用药,IHC弱阳性(++)需行FISH确定有无基因扩增.     

荧光原位杂交与免疫组化法检测乳腺癌HER-2表达的临床意义

[目的]探讨乳腺癌中荧光原位杂交(FISH)检测HER-2基因的扩增和免疫组织化学(IHC)检测HER-2蛋白表达在临床诊断及分子靶向治疗中的应用。[方法]采用FISH与IHC检测50例乳腺癌组织中HER-2基因的扩增与HER-2蛋白表达。[结果]50例浸润性乳腺癌中,FISH检测HER-2基因扩增阳性15例(30%),IHC检测HER-2蛋白表达(-～+)40例,HER-2蛋白表达(++)的2例,HER-2蛋白表达(+++)的8例。[结论]FISH检测HER-2基因的扩增与IHC检测HER-2蛋白表达(++～+++)有较高的一致性。对于IHC检测HER-2(-～++)表达时,必须进一步FISH检测。     

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。     

拓扑异构酶Ⅱa表达与乳腺癌患者生存的相关性

目的 探讨拓扑异构酶Ⅱα(TOP2A)的表达与乳腺癌患者生存的相关性.方法 通过实时定量逆转录聚合酶链反应(quantitative RT-PCR)方法检测86例乳腺癌组织中的TOP2A基因表达量,应用荧光原位杂交(FISH)检测乳腺癌组织中TOP2A基因扩增,应用免疫组化(IHC)法检测TOP2A蛋白表达,并分析TOP2A基因表达量、TOP2A基因扩增与TOP2A蛋白表达的相关性.结果 86例乳腺癌患者中,TOP2A基因扩增11例,TOP2A基因缺失11例,TOP2A基因变异率为25.6%(22/86);TOP2A蛋白表达的阳性率为66.3%(57/86).TOP2A基因表达水平与蛋白表达水平显著相关(P=0.001),TOP2A基因表达水平与乳腺癌患者的无进展生存期显著相关(P<0.001).TOP2A基因扩增与TOP2A蛋白阳件表达无显著相关性(P=0.211).结论 TOP2A的基因定量表达水平为乳腺癌客观可靠的临床预后指标.     

拓扑异构酶Ⅱa表达与乳腺癌患者生存的相关性

目的 探讨拓扑异构酶Ⅱα(TOP2A)的表达与乳腺癌患者生存的相关性.方法 通过实时定量逆转录聚合酶链反应(quantitative RT-PCR)方法检测86例乳腺癌组织中的TOP2A基因表达量,应用荧光原位杂交(FISH)检测乳腺癌组织中TOP2A基因扩增,应用免疫组化(IHC)法检测TOP2A蛋白表达,并分析TOP2A基因表达量、TOP2A基因扩增与TOP2A蛋白表达的相关性.结果 86例乳腺癌患者中,TOP2A基因扩增11例,TOP2A基因缺失11例,TOP2A基因变异率为25.6%(22/86);TOP2A蛋白表达的阳性率为66.3%(57/86).TOP2A基因表达水平与蛋白表达水平显著相关(P=0.001),TOP2A基因表达水平与乳腺癌患者的无进展生存期显著相关(P<0.001).TOP2A基因扩增与TOP2A蛋白阳件表达无显著相关性(P=0.211).结论 TOP2A的基因定量表达水平为乳腺癌客观可靠的临床预后指标.     

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.     

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.     

Protein expression and gene amplification of epidermal growth factor receptor in thymomas

BACKGROUND: Epidermal growth factor receptor (EGFR) overexpression and amplification are important prognostic factors in many solid tumors and anti-EGFR antibody-based therapy is now available as a promising therapeutic modality. There is little information in the literature regarding the biologic role of EGFR in thymomas that are characterized by variable clinical presentations, histologic heterogeneity, and unpredictable behavior. METHODS: Protein expression and gene amplification of EGFR were investigated in 32 thymomas (9 World Health Organization [WHO] type A, 5 type AB, 7 type B2, 7 type B3, 4 type C) using immunohistochemistry and fluorescence in situ hybridization (FISH). FISH analysis included assessment of the average number of copies of the EGFR gene per cell, the average ratio of the EGFR gene to chromosome 7 copy numbers, and ploidy. RESULTS: The results of FISH analysis showed statistically significant correlation with WHO histologic type, invasion, advanced clinical stage, but not with tumor size and outcome. Thymomas associated with myasthenia gravis more frequently showed hyperploidy when compared with sporadic tumors, but there was no difference in EGFR gene amplification. EGFR protein expression assessed by immunohistochemistry did not correlate with any studied clinicopathologic variables. There was poor correlation between the protein expression and gene amplification, only 7 of 23 specimens (30%). CONCLUSIONS: The potential role of EGFR in the pathogenesis of advanced-stage thymomas indicated that evolving anti-EGFR antibody therapy may be considered as a treatment option.     

IHC与FISH检测浸润性乳腺癌患者HER2蛋白表达和基因扩增的差异性分析

目的:探讨并比较免疫组织化学法（IHC）与荧光原位杂交法（FISH）检测浸润性乳腺癌人表皮生长因子受体-2（HER2）蛋白表达和基因扩增的差异性。方法:采用IHC法和FISH法分别检测桂西地区120例乳腺癌患者石蜡标本中HER2蛋白表达与基因扩增情况,比较IHC与FISH检测结果一致性并进行结果相关性分析。对检测不一致的病例重新检测及判读,分析差异原因。结果:120例乳腺癌患者中IHC 33例阳性（3+）中FISH检测阳性33例,阳性符合率100%;IHC 61例不确定（2+）中FISH检测阳性24例,阳性符合率39.34%;IHC 26例阴性（0/1+）中FISH检测阴性21例,阴性符合率80.77%。结果显示,除IHC（2+）外,IHC检测HER2蛋白表达与FISH检测HER2基因扩增有较好的一致性（P＜0.05）。造成两种检测方法差异的原因可能有标本固定不及时,抗体浓度偏低等。结论:IHC法检测HER2与FISH法一致性较好。临床实践中可根据实际情况,结合使用,以便指导临床治疗。     

Characterization of the HER-2/neu oncogene by immunohistochemical and fluorescence in situ hybridization analysis in oral and oropharyngeal squamous cell carcinoma.

PURPOSE: The role of HER-2/neu in squamous cell carcinoma (SCC) of the head and neck is not well defined. The purpose of the current study is to measure the frequency of HER-2/neu expression, to demonstrate HER-2/neu gene amplification in the cases found to be positive for protein overexpression, and to investigate the prognostic significance of overexpression and/or amplification in SCC of the head and neck. EXPERIMENTAL DESIGN: A cohort of 77 patients with SCC of the oral cavity or oropharynx, with stage III or IV disease and uniformly treated with surgical resection and postoperative radiation, served as the primary patient population for the study. Of these, 56 patients had adequate follow-up and paraffin-embedded specimens available for analysis. Median follow-up was 6.1 years. Each of the paraffin-embedded specimens were immunohistochemically stained for HER-2/neu expression and graded for intensity of staining by a pathologist. All cases that demonstrated positive staining by immunohistochemistry were analyzed by fluorescence in situ hybridization (FISH) to assess HER-2/neu amplification status. RESULTS: Five-year survival for the 56 evaluable patients was 40%, with 25% experiencing local relapse, 18% regional relapse, and 25% distant relapse. The percentage of tumors staining positive for HER-2/neu by immunohistochemistry was 17%. There was no statistically significant correlation between HER-2/neu and T stage, N stage, tumor grade, survival, or disease-free survival. HER-2/neu expression did correlate with vascular endothelial growth factor expression. FISH analysis revealed four cases that were amplified for HER-2/neu. Of note, of the 4 amplified cases, 2 suffered regional relapse, 1 suffered distant metastasis, and all 4 expired by 5 years of follow-up. CONCLUSIONS: This is the first demonstration of HER-2/neu gene amplification by FISH in SCC of the head and neck. FISH validates a previously contested controversial role for HER-2/neu gene overexpression in SCC of the head and neck. The prognostic significance and clinical implications of HER-2/neu expression and amplification in head and neck cancer will require additional studies.     

Epidermal growth factor receptor analyses in colorectal cancer: a comparison of methods

EGFR immunohistochemistry (IHC) status is not a reliable predictive marker for response to EGFR-targeted therapies. The present study compares the EGFR status at DNA, RNA and protein level. Blood samples, corresponding normal colon and colorectal cancer tissue were collected from 199 colorectal cancer (CRC) patients. EGFR status was evaluated by FISH analysis, real-time RT-PCR, ELISA and IHC. A polymorphism in the EGFR promoter was evaluated by PCR analysis. The EGFR levels by different methods were mutually compared. Seventy-eight percent of primary tumours and corresponding lymph nodes had equivalent EGFR status (28/34). There was a tendency to higher median protein level (by ELISA) in IHC positive patients compared to IHC negative patients (p=0.086). The median EGFR gene expression level was significantly lower in tumours than in the normal colon with no difference according to IHC status. No tumours had increased gene copy number by FISH. EGFR Sp1-216 polymorphism analysis showed a tendency for different EGFR tumour protein levels and gene expression levels according to the different genotypes. The results show a poor correlation between EGFR status at DNA, RNA and protein level. The predictive value of a combination of methods needs further evaluation in the clinical setting.     

AIB1基因在卵巢癌中的扩增与蛋白表达及其临床意义

目的：探讨AIB1基因在卵巢癌中的基因扩增与蛋白表达及其临床意义。方法：运用荧光原位杂交、免疫组化和流式细胞学方法，对80例卵巢癌组织中AIB1基因的扩增与表达和DNA倍性进行检测，结合肿瘤的临床病理学资料．分析其相关性、结果：80例卵巢癌中，有28例（35％）出现AIB1蛋白的过度表达，而且与肿瘤的Silverberg氏分级和FIGO分期均有显著的相关性（P〈0．05），其中44％的Silverberg氏G2-3级和46％的FIGO Ⅲ～Ⅳ期卵巢癌出现AIB1蛋白的过度表达，其阳性率明显高于G1级（13％）和FIGO Ⅰ～Ⅱ期（14％）的肿瘤；在FISH检测中，9％的卵巢癌出现AIB1基因扩增。另外．89％的AIB1蛋白过度表达的卵巢癌表现为非整倍体的DNA含量。结论：卵巢癌中AIB1蛋白过度表达与肿瘤的病理分级、临床分期和染色体不稳定性密切相关。     

C-myc蛋白在睾丸肿瘤中的表达及意义

目的　探讨C myc基因蛋白与睾丸肿瘤生物学行为及预后的关系。方法　采用免疫组化S P法对 38例睾丸肿瘤和 10例正常睾丸组织中C myc蛋白表达进行了定量分析。结果　 31例 (81.6 % )睾丸肿瘤显示阳性反应 ,正常睾丸组织为阴性反应。肿瘤组和正常组组织阳性细胞率分别为 2 4 .99%和 3.2 6 % ,两者相比有非常显著性差异 (P <0 .0 1)。在睾丸肿瘤中C myc蛋白的表达程度与肿瘤病理分级、临床分期无关 (P >0 .0 5 )。C myc表达在精原细胞瘤组及非精原细胞瘤组间无显著性差异 (P >0 .0 5 )。结论　C myc蛋白的异常表达可能在睾丸肿瘤的发生、发展过程中起着促进作用。