食管鳞癌p53、c-erbB-2蛋白表达研究

为探讨p53、c-erbB-2蛋白表达与食管鳞癌生物学行为的关系,应用免疫组化LSAB法研究181例食管鳞癌中p53、c-erbB-2蛋白的表达。结果发现,正常食管粘膜均无p53、c-erbB-2蛋白的表达。47％食管鳞癌出现p53表达,p53阳性病例癌旁非典型增生上皮出现p53表达,p53阴性病例癌旁非典型增生上皮亦为阴性。p53阳性表达率与患年龄、性别、肿瘤大小、组织学分级,临床TNM分期无关,且与预后无关。51.4%食管鳞癌呈现c-erbB-2蛋白表达,癌旁非典型增生上皮无c-erbB-2表达。c-erbB-2阳性率与肿瘤组织学分级、浸润深度、淋巴结转移及肝转移有关,c-erbB-2阳性表达预后较差。结果提示,p53表达在食管鳞癌发生中起重要作用;c-erbB-2表达在食管鳞癌浸润转移中起重要作用。同时进行p53、c-erbB-2蛋白免疫组化检测,有助于对食管鳞癌进行早期诊断,监测病情和判断预后。     

MACC1在结肠癌中表达及其临床病理意义

目的：观察MAccl（metastasis．associatedincoloncancer-1）在结肠癌中的表达并分析其与大肠癌临床病理特征的关系。方法：选择我院2001年1月～2011年12月收治的317例接受结肠癌根治术治疗的结肠癌患者为研究对象,通过免疫组织化学技术检测MACCl在结肠癌及癌旁正常结肠组织中的表达,并分析MACCl的表达与结肠癌患者临床病理指标和生存期的相关性。结果：MACCl在结肠癌中的阳性表达率选择高于癌旁正常结肠组织（P〈0．05）。MACCl的表达与肿瘤的大小、临床分期、淋巴结转移、远处转移均显著相关（P〈0．05）,MACCl阳性表达病例的肿瘤体积较MACCl阴性表达病例大,临床分期较MACCl阴性表达病例晚,淋巴结转移和远处转移的发生率较MACCl阴性表达者高,均有统计学意义（P〈0．05）。MACCl阳性表达病例的生存率和生存期均较MACCl阴性表达病例显著降低和缩短（P=0．01）。结论：MACCl的表达上调与结肠癌的发生发展密切相关,可能作为评估结肠癌患者预后的参考指标。     

Rspo1、LGR5、NF-κB/p65在胃癌中的表达及其临床意义

目的 通过检测特异性顶部盘状底板反应蛋白1(Rspo1)、富含亮氨酸的重复G蛋白偶联受体5(LGR5)与核转录因子κB/p65(NF-κB/p65)在人胃癌中的表达水平及与临床病理因素及预后之间的关系,并分析三者在胃癌发生发展中发挥的作用。方法 免疫组织化学SP法测定Rspo1、LGR5及NF-κB/p65在115例胃癌组织标本及20例正常组织标本中的表达。结果 Rspo1、LGR5、NF-κB/p65在胃癌中表达较正常组织增高;Rspo1、LGR5、NF-κB/p65的表达与浸润深度、TNM分期、淋巴结及远处转移有关,同时Rspo1表达与肿瘤大小、LGR5表达与肿瘤大小及分化程度有关;胃癌组织中Rspo1与LGR5、NF-κB/p65的表达呈正相关;Kaplan-Meier分析显示Rspo1、LGR5和NF-κB/p65表达阳性组3年生存率均低于阴性组;单因素Cox分析提示肿瘤大小、分化程度、浸润深度、淋巴结转移、远处转移、Rspo1、LGR5、NF-κB/p65阳性是影响胃癌患者预后的危险因素;多因素Cox分析提示淋巴结转移、远处转移、Rspo1及LGR5阳性是影响胃癌患者预后的独立危险因素。结论 Rspo1、LGR5、NF-κB/p65的表达与胃癌的发生发展有关;Rspo1-LGR5在激活Wnt/β-catenin通路的同时可能激活NF-κB通路,二者对胃癌发展有协同作用。     

胃腺癌组织中GDF-15和p53表达特征及其临床意义

目的探讨胃腺癌及癌旁正常组织中生长分化因子15(growth differentiation factor-15,GDF-15)和p53表达的特征及临床意义。方法采用免疫组织化学Polink-1两步法检测91例胃癌手术标本,相应91例癌旁正常组织、64例肠化腺体及36例淋巴结转移癌组织中GDF-15和p53的表达特征,并结合临床资料分析其表达与临床病理参数的关系。结果 GDF-15在肠上皮化生腺体、胃腺癌和淋巴结转移癌组织中的阳性表达率均显著低于癌旁组织。GDF-15的阳性表达率与患者年龄、浸润深度、淋巴结转移及p TNM分期呈正相关,而与其它临床病理参数无关。p53在癌旁正常胃粘膜腺体和肠上皮化生腺体中均不表达,在胃腺癌和淋巴结转移癌组织中的过表达阳性率均高于癌旁组织和肠上皮化生腺体。p53过表达的阳性率与患者年龄、Lauren分型(肠型与弥漫型)和分化程度呈正相关,而与其它临床病理参数无关。GDF-15与p53免疫反应性在胃癌组织中呈正相关。结论 GDF-15可作为胃腺癌的胃壁浸润能力、淋巴结转移能力及临床病理分期的重要评价指标,而p53过表达可能与Lauren分型的肠型胃癌有关。尤其在老年患者中,GDF-15和p53过表达在胃腺癌的发生、进展中扮演重要角色,并起协同作用。     

非小细胞肺癌组织中VEGF与P53蛋白表达及意义

目的:探讨VEGF和P53在非小细胞肺癌(NSCLC)中的表达及其生物学行为的关系。方法:采用免疫组织化学方法,对病理确诊的62例非小细胞肺癌组织进行VEGF和P53表达的检测。结果:62例非小细胞肺癌VEGF阳性表达率58.1%(36/62),其表达与NSCLC的组织分化程度、生存期有相关性(P<0.05),与组织学类型、有无淋巴结转移、临床病理分期无关。P53阳性表达率46.8%(29,62),表达与肿瘤组织类型、TNM分期、有无淋巴结转移无关(P<0.05)。结论:VEGF和P53可作为评价非小细胞肺癌预后的重要指标。用免疫组化检测出P53蛋白可间接反映P53基因的突变。本组资料检测的结果显示,P53基因阳性表达率46.8% (29/62),表明近一半肺癌病例已失去P53蛋白的抑癌功能,且P53基因的突变可能与非小细胞肺癌的发生有关。P53和VEGF都是潜在的新型肿瘤表型标志物,它们的检测可作为肿瘤病理分级诊断的主要参考依据。同时检测P53和VEGF的表达状态,可通过肿瘤血管形成的生物学行为信息,进一步研究肿瘤血管形成的分子水平调控机制,这有助于抗肿瘤药物的开发研究。     

Beclin1与p53在唾液腺多形性腺瘤和癌在唾液腺多形性腺瘤中的表达及与临床病理因素的关系研究

目的:探讨自噬体相关基因Beclin1与p53基因在唾液腺多形性腺瘤(PA)和癌在唾液腺多形性腺瘤(CPA)中的表达及其与临床病理因素的关系。方法:选择2013年1月至2017年8月于我院手术切除的PA标本45例作为PA组,CPA标本32例作为CPA组,正常腮腺组织30例作为对照组,应用免疫组化SP法对三组Belin1与p53基因表达情况进行检测,分析CPA组Beclin1、p53表达情况与临床病理因素关系及Beclin1、p53表达的相关性。结果:三组Beclin1、p53阳性表达率存在统计学差异,CPA组Beclin1阳性表达率低于PA组和对照组,CPA组和PA组p53阳性表达率高于对照组,且CPA组高于PA组(P0.05)。CPA组Beclin1、p53表达情况与TNM分期、淋巴结转移有关,TNM分期为III-IV期患者Beclin1阳性表达率低于I-II期患者,p53阳性表达率高于I-II期患者,淋巴结转移患者Beclin1阳性表达率低于无淋巴结转移患者,p53阳性表达率高于无淋巴结转移患者,差异均有统计学意义(P0.05),CPA组Beclin1、p53表达情况与年龄、性别、肿瘤直径、侵袭性无关(P0.05)。经Spearman相关性分析可得,CPA患者Beclin1和p53的表达呈负相关关系(r=-0.839,P=0.000)。结论:PA及CPA中存在Beclin1异常低表达,p53异常高表达,CPA患者Beclin1、p53表达情况与TNM分期、淋巴结转移有关,且两者表达呈负相关。     

乳腺癌中癌胚抗原（CEA）与p53、nm23—H1蛋白表达和部分病理指标的相关性研究

利用免疫组织化学方法检测了乳腺癌CEA与p53和nm23－H1蛋白的表达,对其相关性进行了研究,同时与其它病理指标亦进行了比较。结果：82 例乳腺癌中,CEA阳性67例（82％）,与p53蛋白的表达呈显的负相关（P＜0.05）,而与nm23-H1蛋白的表达则呈显的正相关（P＜0.05）;同时,CEA的表达与肿瘤的病理分级和体积显相关（P＜0.05）,即分级越高或肿瘤越大,CEA阳性表达率越低;而与患年龄、淋巴结转移和肿瘤坏死程度无关（P＞0.05）。综合分析推测：CEA可能是乳腺癌一种高分化肿瘤标志,并与肿瘤的浸润转移潜能有一定的关系,与其它指标联合应用在判断乳腺癌预后有一定的价值。     

Nucleobindin-2 在乳腺癌中表达的临床病理意义

目的:探讨NUCB2在乳腺癌中的表达及临床病理意义。方法:收集乳腺癌病例及相应的临床资料包括随访资料应用免疫组织化学技术检测良性病变,有/无淋巴结转移的乳腺癌及配对淋巴结转移灶中NUCB2的表达,分析NUCB2表达与乳腺癌临床病理指标和生存状态间的关系。结果:免疫组化结果:显示NUCB2在BBD、MBC、NMBC、PMLN的阳性表达率分别为90.0%(45/50),82.1%(87/106)、48.2%(40/83)、47.2%(50/106)。其中,NUCB2在BBD的阳性表达率显著高于NMBC、PMLN,而与MBC无显著性差异;在MBC中的阳性表达率显著高于NMBC;在MBC中的阳性表达率显著高于PMLN。临床病理分析结果:显示乳腺癌原发灶中NUCB2表达与淋巴结转移(p=0.000)、临床分期(p=0.001)、雌激素阳性表达(p=0.020)具有显著性相关。而与年龄、肿块大小、组织学分级、组织学类型、孕激素受体、HER2表达、绝经情况无显著相关。Kaplan-Meier分析结果:显示NUCB2阳性表达病例的总生存率比阴性病例更短(P=0.004)。结论:NUCB2在乳腺癌中表达与淋巴结转移、临床分期、雌激素受体表达和生存状态相关,可能在乳腺癌恶性演进和预后中具有重要意义。     

口腔鳞状细胞癌中hMSH2、PCNA和p53的表达及意义

目的:探讨hMSH2、PCNA和p53在OSCC中的表达及可能存在的临床意义.方法:运用免疫组织化学S-P法对56例OSCC组织中hMSH2、PCNA和p53的表达进行检测.结果:(1)3种基因产物在OSCC中的阳性率均高于正常口腔黏膜,中-低分化癌的阳性率高于高分化癌,有淋巴结转移的阳性率高于无淋巴结转移.(2)hMSH2与PCNA、p53,p53与PCNA的表达均呈正相关.(3)5年生存率与hMSH2蛋白表达和淋巴结转移有关.结论:hMSH2、PCNA和p53的异常表达及其相互之间的调节可能与OSCC的发生发展有关.     

涎腺粘液表皮样癌中hMLH1、hMSH2与p53基因的表达及意义

目的:探索hMLH1、hMSH2基因在涎腺粘液表皮样癌中的表达及其与p53表达的关系.方法:采用免疫组织化学S-P法对涎腺粘液表皮样癌及正常涎腺组织石蜡切片进行hMLH1、hMSH2及p53表达的检测.结果:(1)hMLH1、hMSH2及p53在癌组织中表达阳性率均显著高于正常组织(P＜0.01).(2)hMLH1、hMSH2的表达与癌的分化程度、患者年龄、性别、肿块大小、浸润深度以及有无淋巴结转移均无关(P＞0.05).(3)hMLH1、hMSH2基因均与p53表达呈正相关.结论:检测hMLH1、hMSH2、p53将有助于判断涎腺粘液表皮样癌的恶性程度和相关的生物学行为.     

110例维吾尔族及汉族乳腺癌中BRCA1基因突变及P53蛋白表达的研究

目的研究维吾尔族及汉族乳腺癌患者BRCA1基因突变及P53蛋白的表达。方法选取70例维吾尔族和40例汉族乳腺癌根治标本,对照组为32例维汉族乳腺良性病变(纤维腺病及纤维腺瘤)及乳腺癌旁非癌组织;运用PCR-SSCP和DNA序列测定的方法检测BRCA1基因突变及Evision二步法检测P53蛋白的表达。结果(1)110例维吾尔族及汉族乳腺癌中发现BRCA1突变的14个新位点。(2)110例维吾尔族及汉族乳腺癌BRCA1的突变率为10%,其中22例维吾尔族早发性乳腺癌(≤35岁)BRCA1突变率为31.82%,高于维吾尔族晚发性乳腺癌(P<0.01)。(3)110例维吾尔族及汉族乳腺癌中发现11例BRCA1基因核苷酸多态性位点。(4)BRCA1基因突变相关性乳腺癌中P53蛋白阳性表达率高于对照组,其淋巴结转移率高于对照组,其发病年龄小于对照组(P<0.01)。结论BRCA1基因突变与新疆维吾尔族早发性乳腺癌密切相关,且BRCA1突变相关性乳腺癌具有P53阳性率高、发病年龄趋于年轻化、淋巴结转移率高的趋势,这些特点有可能为基因检测前的筛选提供参考依据。     

p53 mutations as tumor markers in fine needle aspirates of palpable breast masses

OBJECTIVE: Mutations in p53 exons 5-8 are found in 40-50% of breast carcinomas. We performed a retrospective analysis of p53 mutations in fluid-based, archival fine needle aspirates (FNAs) of breast masses to determine their potential diagnostic utility as breast tumor cell markers. STUDY DESIGN: Residual, fluid-based, archival FNAs of 27 breast masses were retrospectively evaluated by polymerase chain reaction (PCR), single-strand conformational polymorphism analysis (SSCP) and sequencing for p53 exons 5-8. Results were compared with the morphologic diagnoses and genotyping of available excisional biopsy tissue. RESULTS: Six of the twenty-seven cases were found to have a clonal mutation in p53; all six mutated cases showed carcinoma on subsequent excisional biopsy. Definitive cytologic diagnosis of cancer had been possible in only four of the six cases. Identical mutations were found in the excised carcinomas in the five cases with available tissue. None of the 14 aspirates with benign cytology had detectable mutations in p53. CONCLUSION: p53 Mutational analysis by PCR/SSCP/sequencing deserves to be critically studied as a diagnostic criterion in patients with indeterminate or suspicious cytology. Validation studies should be performed to test p53 mutations as molecular diagnostic markers in breast cytology specimens.     

Detection of K-ras and p53 mutations in sputum samples of lung cancer patients using laser capture microdissection microscope and mutation analysis

Mutations in the p53 tumor suppressor gene and the K-ras oncogene have been frequently found in sputum and bronchoalveolar lavage (BAL) samples of lung cancer patients and other patients prior to presenting clinical symptoms of lung cancer, suggesting that they may provide useful biomarkers for early lung cancer diagnosis. However, the detection of these gene mutations in sputum and BAL samples has been complicated by the fact that they often occur in only a small fraction of epithelial cells among sputum cells and, in the case of p53 gene, at many codons. In this study, sputum cells were collected on a filter membrane by sputum cytocentrifugation and morphologically analyzed. Epithelial cells were selectively taken by using a laser capture microdissection microscope and analyzed by polymerase chain reaction (PCR) and single-stranded conformational polymorphism (SSCP) for p53 mutations and by PCR and denaturing gradient gel electrophoresis (DGGE) for K-ras mutations. This method was used to analyze sputum of 15 Chinese women with lung cancer from Xuan Wei County, China and detected mutations in sputum of 7 (46.7%) patients, including 5 patients with p53 mutations, 1 patient with a K-ras mutation, and 1 patient with K-ras and p53 mutations. For comparison, only two of the mutations were detected by conventional methods. Therefore, the laser capture/mutation analysis method is sensitive and facilitates the detection of low-fraction mutations occurring throughout the p53 and K-ras genes in sputum of lung cancer patients. This method may be applicable to the analysis of epithelial cells from clinically normal sputum or BAL samples from individuals with a high risk for developing lung cancer.     

Multiple mutations of the p53 gene in human mammary carcinoma

Alteration of the p53 tumor suppressor gene is the most common genetic abnormality in human cancer. In breast cancer, depending on the stage of disease and method of detection, mutation rates of 25-60% have been observed. Multiple mutations of p53 gene in the same tumor however, are rarely reported. In this study we explored the frequency of multiple mutations of p53 gene in mammary carcinoma in a cohort of south Florida patients. Three hundred eighty-four cases of primary breast cancer diagnosed between 1984 and 1986 at the University of Miami, Jackson Medical Center were subjects of this study. Sequence analysis of exons 5 through 8 of p53 was performed on cloned PCR-amplified DNA of formalin-fixed, paraffin-embedded tumors. Two hundred thirty-four of 384 breast cancers (61%) had p53 mutation. Of those, 36 tumors showed more than one mutation; 31 tumors had two mutations, three showed three, one tumor had five mutations, and one case carried six mutations. The majority of mutations were missense (43) followed by silent (35); and most occurred within a single exon. Our study suggests that multiple mutations of p53 suppressor gene in breast cancer are more common than currently believed.     

FISH和IHC检测HER2基因及蛋白在乳腺癌诊治中的应用

目的对比研究免疫组织化学(IHC)与荧光原位杂交(FISH)方法检测乳腺癌中C-erbB-2蛋白表达和HER2基因扩增,评估两种方法的实际应用价值。方法 IHC和FISH法分别检测58例乳腺癌组织中C-erbB-2蛋白表达和HER2基因扩增状况,并进行一致性分析。结果IHC检测蛋白阳性2+和3+共22例,占总病例的37.9%,58例乳腺癌中FISH检测出HER2基因扩增17例,占29.3%。IHC检测C-erbB-2蛋白3+的12例中11例HER2基因扩增阳性,1例无扩增;C-erbB-2蛋白2+者10例,其中5例可见HER2基因扩增;C-erbB-2蛋白1+者11例HER2基因扩增仅1例。C-erbB-2蛋白阴性病例共25例均未检测到基因扩增。结果显示IHC检测C-erbB-2蛋白与FISH检测HER2基因扩增状态有较高的一致性(k=0.681,P0.01),C-erbB-2蛋白阴性和3+标本与HER2基因扩增阳性率比较差异无统计学意义(P0.05);而C-erbB-2蛋白1+和2+病例与其HER2基因扩增差异具有统计学意义(P0.05)。结果 IHC检测C-erbB-2蛋白强阳性(3+)和阴性(0)与HER2基因扩增有较高的一致性,可作为临床是否应用Herceptin治疗的依据,而C-erbB-2蛋白2+和1+病例必须进一步行HER2基因扩增检测。     

Comparison of multiplex ligation dependent probe amplification to immunohistochemistry for assessing HER-2/neu amplification in invasive breast cancer.

The HER-2/neu transmembrane tyrosine kinase receptor is both a prognostic marker and a therapeutic target for breast cancer. Accurate determination of HER-2/neu status is a prerequisite for selecting breast tumors for HER-2/neu immunotherapy or for taxan based chemotherapy. Unfortunately, there is no consensus concerning how this determination should be reached. We compared assessment of HER-2/neu status using Multiplex ligation-dependent probe amplification (MLPA) and immunohistochemistry (IHC). The patient group comprised 60 Indonesian breast cancers patients. IHC was performed on paraffin sections using the CB11 antibody from Novocastra. Results were scored according to the Hercept test. For MLPA, DNA was extracted from frozen samples, PCR amplified with a probe set containing three hemi-primer sets for the HER-2 locus and another nine control probes spread over chromosome 17 and other chromosomes, and analyzed on a gene scanner. A ratio above two for at least two HER-2 locus probes compared to the control probes was regarded as amplification. IHC for HER-2/neu was negative in 36 cases, and 24 cases (40%) showed expression. Seven, eight and nine of the latter cases were 1+, 2+ and 3+ positive, respectively. Forty-seven cases showed no amplification by MLPA, and 13 cases (22%) were amplified. Comparison of IHC and MPLA showed that none of the 36 IHC-negative or seven IHC 1+ cases was amplified. Five of the eight (63%) 2+ cases were amplified, and eight of nine (89%) of the IHC 3+ tumors showed gene amplification by MLPA assay. For HER-2/neu, there is a good correlation between gene amplification detected by MLPA and overexpression by IHC in invasive breast cancer. It appears that MLPA can detect the HER-2 amplified cases in the IHC 2+ class. Because MLPA is quick and inexpensive, it is an attractive method for detecting HER-2/neu amplification in daily laboratory practice.     

Infrequent but significant p21/WAF1/CIP1 gene alteration in synchronous or metachronous transitional cell carcinoma

p53 inducible cyclin dependent kinase inhibitor, p21/WAF1/CIP1(p21), played a pivotal role for G1 arrest when cells received genotoxic stimuli. p21 could be a putative tumor suppressor gene, since its dysfunction may lead to accumulation of genomic alteration. We investigated the p21 and p53 status using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and immunohistochemical analyses, in eight patients who had synchronous or metachronous urothelial tumors. Loss of heterozygosity (LOH) of p21 gene was detected in one coincidental tumor in one case. p21 positive cells were detected by immuno-histochemical staining in all tumors in one case, and in one coincidental tumor in two cases. Among p21 positive cells in these three cases, no p53 mutations were detected, whereas no p21 positive cells were detected in other cases with a p53 mutation. These findings suggested that in transitional cell carcinoma (TCC) p21 gene mutation is infrequent like the p53 gene mutation, but that LOH might be important in the inactivation of p21.     

人胃癌细胞系中p53抑癌基因变异的检测及其序列分析

ｐ５３抑癌基因由于点突变,缺失或易位等方式而丧失活性是多种肿瘤发生发展的重要机理之一。本文应用聚合酶链式反应─—单链构象多态性分析（ＰＣＲ－ＳＳＣＰ）方法,对四种人胃癌细胞系ＭＧＣ８０３,ＢＧＣ８２３,ＧＣ７９０１和ＰＡＭＣ－８２中ｐ５３基因的第５、６、７、８四个外显子进行检测,结果发现,ＰＡＭＣ－８２的第５和第８外显子,ＧＣ７９０１的第６外显子存在突变。ＰＣＲ－直接测序证明,它们分别在１７４位、２８０位、２０４位密码子发生缺失Ｇ,ＧＣ→ＣＧ,ＡＴ→ＣＧ转变,因而可使其编码的ｐ５３蛋白分别发生移码突变,Ａｒｇ→Ｔｈｒ,Ｇｌｕ→Ａｌａ,而丧失肿瘤抑制功能。实验结果表明,ｐ５３基因在胃癌发生发展过程中,尤其是较晚阶段具有重要作用。     

P_(53)突变、表达及人乳头状瘤病毒感染与宫颈癌

利用免疫组化、聚合酶链反应(PCR)、单链构象多态性(SSCP)分析等方法, 对49例同一标本宫颈癌组织中p53蛋白、P53外显子7～8变异、HPV6、11、16、18-DN A进行检测,以探讨它们在宫颈癌形成中的作用、相互关系和临床意义.结果表明: a.P5 3基因外显子7～8突变率14.29％、p53蛋白阳性率48.98％、HPV-DNA阳性率87.76％.b.P53基因突变不一定伴有p53蛋白阳性,但P53基因突变而p53蛋白阴性的标 本必是HPV-DNA阳性;91.67％的p53蛋白阳性标本具有HPV-DNA阳性.c.HPV16-DNA阳 性率显著高于HPV6、11、18-DNA阳性率.证明：宫颈癌的发生主要与HPV16感染有关,其 次是P53基因突变所致;p53蛋白阳性由HPV感染和/或P53基因突变所致.     

Comparison of multiplex ligation dependent probe amplification to immunohistochemistry for assessing HER-2/neu amplification in invasive breast cancer

The HER-2/neu transmembrane tyrosine kinase receptor is both a prognostic marker and a therapeutic target for breast cancer. Accurate determination of HER-2/neu status is a prerequisite for selecting breast tumors for HER-2/neu immunotherapy or for taxan based chemotherapy. Unfortunately, there is no consensus concerning how this determination should be reached. We compared assessment of HER-2/neu status using Multiplex ligation-dependent probe amplification (MLPA) and immunohistochemistry (IHC). The patient group comprised 60 Indonesian breast cancers patients. IHC was performed on paraffin sections using the CB11 antibody from Novocastra. Results were scored according to the Hercept test. For MLPA, DNA was extracted from frozen samples, PCR amplified with a probe set containing three hemi-primer sets for the HER-2 locus and another nine control probes spread over chromosome 17 and other chromosomes, and analyzed on a gene scanner. A ratio above two for at least two HER-2 locus probes compared to the control probes was regarded as amplification. IHC for HER-2/neu was negative in 36 cases, and 24 cases (40%) showed expression. Seven, eight and nine of the latter cases were 1+, 2+ and 3+ positive, respectively. Forty-seven cases showed no amplification by MLPA, and 13 cases (22%) were amplified. Comparison of IHC and MPLA showed that none of the 36 IHC-negative or seven IHC 1+ cases was amplified. Five of the eight (63%) 2+ cases were amplified, and eight of nine (89%) of the IHC 3+ tumors showed gene amplification by MLPA assay. For HER-2/neu, there is a good correlation between gene amplification detected by MLPA and overexpression by IHC in invasive breast cancer. It appears that MLPA can detect the HER-2 amplified cases in the IHC 2+ class. Because MLPA is quick and inexpensive, it is an attractive method for detecting HER-2/neu amplification in daily laboratory practice.