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.

Abstract：

Objective The aim of this study was to assess the TOP2A RNA expression and die relationship of TOP2A protein expression with metastasis-free interval in breast cancer patients. Methods TOP2A expression was analyzed prior to surgery in 86 patients. The level of TOP2A gene amplification was analyzed by fluorescence in situ hybridization (FISH) , its RNA expression level with RT-PCR, and their correlation with TOP2A protein expression was assessed by immunohistochemistry (IHC). The correlation between RNA expression level and metastasis-free interval in breast cancer patients was also analyzed. Results Aberrations (amplification or deletion) of TOP2A copy number was observed in 25.6% (22/86) of the cases. TOP2A protein expression was detected in 66. 3% (57/86) of the samples. There was a significant correlation between the TOP2A RNA expression and protein expression (P<0.001). TOP2A gene expression was significantly associated with the metastasis-free interval in the breast cancer patients (P =0.001). There was no significant correlation between TOP2A gene amplification and TOP2A protein expression (P=0.211). Conclusions TOP2A RNA level is an objective and reliable prognostic indicator in breast cancer.     

乳腺癌组织TOP2A和HER2/neu扩增与临床病理因素相关性研究

目的 拓扑异构酶Ⅱ(typeⅡtopoisomerase,TOP2A) DNA是常见的化疗疗效预测因子,HER2是与乳腺癌相关的重要的原癌基因之一.本研究探讨乳腺癌组织中人类表皮生长因子受体HER2/neu和TopoⅡ之间的关系,及其与临床病理因素之间的相关性.方法 收集广西医科大学附属肿瘤医院2010-02-01-2012-09-30手术治疗的96例乳腺浸润性导管癌标本,实时定量聚合酶链式反应(real time polymerase chainreaction,RQ-PCR)检测和评估基因扩增水平,免疫组织化学(immunohistochemistry,IHC)微阵列(n=76)检查基因扩增和蛋白质表达水平之间是否存在相互关系.结果 根据RQ-PCR或IHC微阵列取得的HER2/neu基因状态,TOP2A基因的扩增水平差异无统计学意义,P值分别为0.481和0.935.在HER2/neu(-)基因型患者中,29.1％(14/48)的患者显示出TOP2A基因水平高于第三四分位值,然而22.9％(11/48)的HER2/neu(+)基因型患者的数值处于第一四分位值(log TOP2A＜0.62),因此表明存在低水平的扩增.采用IHC以及荧光原位杂交(fluorescence in situ hybridization,FISH)方法确定具有HER2/neu-基因型的60例患者中,22.9％(11/48)的患者在IHC微阵列上被归类为TOP2A(+)基因型患者.同时,采用IHC以及FISH法将患者视为HER2/neu(+)基因型患者的14例患者中,大多数患者(n=10)被归类为TOP2A(+)基因型患者.结论 乳腺癌组织中TOP2A基因的扩增并不局限于带有HER2/neu(+)基因型的患者,并且很大比例的HER2/neu(-)基因型患者表现出具有高水平的TOP2A基因.     

Ki-67、Her-2及Top2A在乳腺癌中的扩增表达与蒽环类药物疗效关系的研究

目的 探讨Ki-67、Her-2及Top2A在乳腺癌中的扩增表达与蒽环类药物疗效的关系.方法 207例经病理证实的乳腺癌患者均接受FEC方案新辅助化疗,并测定其Ki-67、Her-2及Top2A基因的表达.结果 207例乳腺癌患者中,Ki-67分级为(-)32例,(+)58例,(++)95例,(+++)22例;Her-2基因扩增26例(12.6％),未扩增181例(87.4％);Top2A基因扩增26例(12.6％),未扩增181例(87.4％).Ki-67分级与Her-2基因、Top2A基因扩增无关(P＞0.05).Her-2、Top2A的扩增与化疗疗效均呈正相关(r=0.59、0.58,P＜0.05).结论 Her-2、Top2A的表达对乳腺癌患者化疗药物的选择及预后的判断具有重要的指导价值.     

乳腺癌组织中DNA甲基化转移酶与多药耐药基因ABCG2表达的关系

背景与目的:研究乳腺癌组织中DNA甲基化转移酶(DNA methyltransferase,DNMT)与多药耐药基因ABCG2表达的关系,以进一步探讨ABCG2表达的表观遗传学机制.材料与方法:用实时定量RT-PCR法检测22例乳腺癌及其匹配的癌旁组织中DNMT1、DNMT3A、DNMT3B和ABCG2的表达.并采用Spearman等级相关分析DNMTs与ABCG2基因表达的相关性.结果:乳腺癌组织中ABCG2、DNMT1、DNMT3A、DNMT3B mRNA表达量显著高于癌旁组织(P<0.01),并且DNMT3B表达量显著高于DNMT1和DNMT3A(P<0.01),与ABCG2基因表达呈负相关性(r=-0.664,P<0.01). 结论:乳腺癌组织中DNMT3B可能参与了ABCG2基因的表达调控,这为寻找药物靶点并逆转其介导的药物耐受提供了新的科学依据.     

FATS在乳腺癌组织中的表达及临床相关性研究

目的:研究FATS在乳腺癌以及癌旁正常组织的mRNA水平,探讨FATS在乳腺癌发生发展中的作用.方法:本研究通过实时定量PCR技术分析检测天津医科大学附属肿瘤医院的156例乳腺癌患者FATS基因表达水平,并且研究其表达水平与患者临床预后的关系.结果:实时定量PCR的结果显示乳腺癌患者肿瘤组织FATS表达量明显低于其配对的癌旁正常腺体组织(P=0.003).FATS基因低表达与高的组织学分级相关(P=0.01),FATS基因低表达的患者无病生存期有统计学意义(P=0.036).Cox多因素分析显示FATS基因的表达是乳腺癌独立的预后因素(OR=0.549;95%:0.308～0.977;P=0.041).结论:FATS低表达与乳腺癌的发生、发展具有高度相关性,FATS表达是乳腺癌的独立预后因素,有望成为新的肿瘤标记物,为临床诊治提供新的靶点.     

多药耐药基因MDR1及P-gp蛋白在乳腺癌组织中的表达及临床意义

目的检测乳腺癌组织中多药耐药基因MDR1及P-gp蛋白的表达,并探讨其表达与临床病理因素的关系及MDR1与P-gp的相关性。方法采用荧光定量RT-PCR(FQ-RT-PCR)法检测61例乳腺癌、22例对照组(包括11例乳腺良性疾病、11例癌旁正常组织)中MDR1基因的表达,同时采用免疫组化法检测上述标本中P-gp蛋白的表达。结果乳腺癌组织中MDR1基因扩增率为50.82%(31/61),与正常对照组相比差异有统计学意义,P=0.0006。乳腺癌中P-gp蛋白阳性率为42.62%(26/61),其表达与年龄、肿瘤大小、淋巴结转移、ER、PR、月经状况无关;MDR1基因扩增率高于P-gp蛋白表达,二者呈高度相关,但并不完全相符。结论乳腺癌中存在多药耐药基因MDR1及P-gp蛋白的表达,且其表达呈正相关。检测上述基因及其蛋白的表达,可预测乳腺癌的多药耐药。     

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.     

Caveolin-1在乳腺浸润性导管癌间质内癌相关成纤维细胞中的表达水平与乳腺癌分子亚型的相关性

目的 检测Caveolin-1（Cav-1）在乳腺浸润性导管癌间质内癌相关成纤维细胞（CAFs）中的表达,分析Cav-1与乳腺癌分子亚型、HER-2基因状态的相关性及其与预后的关系。方法应用免疫组织化学法检测168例乳腺癌组织Cav-1在CAFs中的表达情况。用荧光原位杂交（FISH）方法检测乳腺癌组织中HER-2基因的扩增状态。结果 Cav-1在HER-2过表达型和Luminal B型的阳性表达率均为83.3％,显著高于Luminal A型（58.1％）和Basal like型（35.3％）,差异有统计学意义（P＜0.05）。乳腺癌HER 2基因扩增占39.3％（66／168）,与HER-2蛋白表达呈正相关（r=0.625,P＜0.05）。HER-2基因扩增组的Cav-1阳性表达率为83.3％,显著高于未扩增组的55.9％（P＜0.05）。Cav-1表达水平与HER-2蛋白（r=0.253）及基因（r=0.305）表达均呈正相关（P＜005）,与预后亦明显相关（P=0.041）。结论 乳腺癌间质内Cav-1表达与分子分型相关,HER 2蛋白表达水平增高及基因扩增与间质Cav-1表达水平增高呈正相关（P＜0.05）,乳腺癌间质CAFs中Cav-1高表达提示患者预后较好。     

TOPⅡα、TP53和HER2在胃腺癌中的表达及其临床意义

目的 检测胃腺癌组织中TOPⅡα、TP53和HER2的表达,探讨其表达的相关性及与胃腺癌患者临床病理特征的关系。方法 免疫组织化学法检测136例胃腺癌及20例正常胃黏膜石蜡标本中TOPⅡα、TP53和HER2蛋白的表达,SPSS20.0软件分析三者之间的相关性及与胃腺癌患者临床病理特征的关系。 结果 （1）136例胃腺癌标本中,TOPⅡα和TP53的阳性表达率分别是47.1%（64/136）和65.4%（89/136）,HER2阳性扩增率为23.5%（32/136）;（2）TOPⅡα和P53的表达均与胃腺癌分化程度及Lauren分型显著相关（P<0.05）,HER2的阳性扩增与肿瘤的分化、Lauren分型以及临床TNM分期显著相关（P<0.05）;（3）TP53同TOPⅡα及HER2均有显著相关性（r=0.326, P=0.000; r=0.212, P=0.012）。结论 联合检测胃腺癌组织中TOPⅡα、TP53和HER2蛋白表达有利于评估胃腺癌的生物学特性和预测患者的预后。     

Gene expression of topoisomerase II alpha (TOP2A) by microarray analysis is highly prognostic in estrogen receptor (ER) positive breast cancer

Introduction Overexpression of Topoisomerase II alpha (TOP2A) has been implicated with gene amplification of the 17q21 amplicon and consecutively with ErbB2 overexpression and amplification. However, gene amplification does not necessarily correlate with RNA and protein expression. There is growing evidence that TOP2A protein expression is a strong prognostic and TOP2A gene amplification might be a predictive marker (particularly for the use of anthracyclines). Methods Large scale analysis was performed using Affymetrix microarray data from n = 1,681 breast cancer patients to evaluate TOP2A expression. Results TOP2A expression showed a strong correlation with tumor size (χ²-test, P < 0.001), grading (P < 0.001), ErbB2 (P < 0.001) and Ki67 expression (P < 0.001) as well as nodal status (P = 0.042). Survival analysis revealed a significant prognostic value in ER positive (n = 994; log rank P < 0.001), but not in ER negative breast cancer patients (n = 369, P = 0.35). The prognostic impact of TOP2A expression was independent of Ki67 expression in ER positive tumors (P = 0.002 and P = 0.007 for high and low Ki67, respectively). Moreover a worse prognosis of high TOP2A expressing tumors was found in the subgroup of ErbB2 negative tumors (P < 0.001) and a trend among ErbB2 positive tumors (P = 0.11). The prognostic value of TOP2A was independent of whether the patients were untreated or had received adjuvant therapy. In multivariate Cox regression analysis including standard parameters TOP2A emerged to be the top prognostic marker (HR 2.40, 95% CI 1.68–3.43, P < 0.001). Conclusion TOP2A expression is an independent prognostic factor in ER positive breast cancer and could be helpful for risk assessment in ER positive breast cancer patients. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. A. Rody and T. Karn contributed equally to this work.     

TOP2A amplification in breast cancer is a predictive marker of anthracycline-based neoadjuvant chemotherapy efficacy

Anthracycline is a DNA topoisomerase 2-α (TOP2A) inhibitor and its concomitant over expression with Human Epidermal Growth Factor Receptor 2 (HER2) was investigated of being predictive for the response to anthracycline-based chemotherapies in breast cancer. 309 early and local advanced breast cancer patients were treated with anthracycline-based neoadjuvant chemotherapies in intense dose dense (IDD) (CE, Cyclophosphamide?+?Epirubicin) or conventional (TE, Paclitaxel?+?Epirubicin) regimens. HER2 proteins were qualitatively analyzed by immunohistochemistry (IHC) of primary tumor core biopsies, and TOP2A gene amplification levels of HER2 over-expressing cases were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). Overall pathological complete response rate (pCR) was achieved in 14.3?%. HER2 was over expressed in 80/309 (25.9?%) cases, of which 61/80 cases have been tested for their TOP2A status. Over expression of HER2 was significantly positively correlated with higher pCR rates compared to low HER2 expression (27.5?% vs. 9.6?%, P?相似文献   

Aberrations of ERBB2 and TOP2A genes in breast cancer

Copy number changes in TOP2A have frequently been linked to ERBB2 (HER2) amplified breast cancers. To study this relationship, copy number changes of ERBB2 and TOP2A were investigated by fluorescence in situ hybridization (FISH) in two cell lines; one characterized by having amplification of both genes and the other by having amplification of ERBB2 and deletion of TOP2A. The characteristics are compared to findings on paired ERBB2 and TOP2A data from 649 patients with invasive breast cancer from a previously published biomarker study. The physical localization of FISH signals in metaphase spreads from cell lines showed that simultaneous amplification is not a simple co‐amplification of a whole amplicon containing both genes. Most gene signals are translocated to abnormal marker chromosomes. ERBB2 genes but not TOP2A genes are present in tandem amplicons, leading to a higher ERBB2 ratio. This observation was confirmed by patient FISH data: among 276 (43% of all patients) abnormal tumors, 67% had different ERBB2 and TOP2A status. ERBB2 amplification with normal TOP2A status was found in 36% of the abnormal tumors (15% of all patients). Simultaneous amplification of both genes was found in 28% of the abnormal tumors (12% of all patients) while TOP2A deletion and ERBB2 amplification was observed in 16% of the abnormal cases (8% of all patients). A small number of tumors had TOP2A amplification (4%) or deletion (6%) without simultaneous changes of the ERBB2 gene. ERBB2 deletion was also observed (5%) but only in tumors with simultaneous TOP2A deletion. The average gene/reference ratio was significantly different: 5.0 for TOP2A but 7.2 for ERBB2 in the amplified tumors (P<0.01). Amplification of the two genes may be caused by different mechanisms, leading to higher level of amplification for ERBB2 compared to TOP2A. In the majority of breast cancer patients, simultaneous aberration of ERBB2 and TOP2A is not explained by simple co‐amplification.     

Topoisomerase 2A gene amplification in breast cancer. Critical evaluation of different FISH probes

The HER2 amplicon on chromosome 17q is variable in size and occasionally includes Topoisomerase 2A (TOP2A) at 17q21-22. It has been suggested that TOP2 co-amplification, not HER2 amplification on chromosome 17q11.2-12, is a useful predictive marker of response to anthracycline-based chemotherapy in breast cancer patients. Given the significant toxicities of anthracyclines, the detection methods of TOP2A gene amplifications have to be standardized. We determined TOP2A gene alterations using two different fluorescence in situ hybridization (FISH) DNA probes. HER2 amplifications were identified with the PathVysion probe. TOP2A status of 42 HER2 amplified breast cancers was tested by FISH with PathVysion covering 160 kb and DAKO pharm DX covering 228 kb of the TOP2A amplicon. TOP2A protein expression was tested by immunohistochemistry. Multiplex-ligation dependent probe amplification (MLPA) was performed retrospectively in cases showing discrepancies. TOP2A was amplified in 15 of 42 cases (35%) with DAKO pharm DX and in 11 of 42 cases (26%) with PathVysion. In all four discrepant cases, MLPA showed no TOP2A amplification, but instead amplification of an upstream region including HER2. TOP2A was deleted in the same seven of 42 carcinomas (17%) with both probes. TOP2A protein expression was detected in all 42 tumours (100%) with high intratumoral heterogeneity. TOP2A amplification rate depends on the length of the hybridized probes for the TOP2A locus. Because TOP2A, not HER2, is a target of anthracyclines, non-overlapping DNA probes should be used to evaluate any associations between such alterations and response to anthracycline-based chemotherapy.     

TOP2A RNA expression and recurrence in estrogen receptor-positive breast cancer

The purpose of this study is to evaluate the relationship between TOP2A RNA expression and recurrence in patients with operable estrogen receptor (ER)-positive breast cancer. We evaluated TOP2A expression in a pooled analysis of four independent datasets with gene expression data including 752 patients with early stage, ER-positive, HER2-negative breast cancer, most of whom received either no adjuvant therapy or only endocrine therapy without chemotherapy. We also used an algorithm to simulate the Oncotype DX Recurrence Score (simRS) and the proliferation component of the Recurrence Score (simPS). Results are expressed as the hazard ratio (HR) for estimates of the effect of a 1SD increase in the value of the log gene expression (x + 1SD vs. x) as a continuous function. TOP2A expression was significantly associated with recurrence (HR 1.56, p < 0.0001), and after adjustment for simRS (HR 1.26, p = 0.003). TOP2A correlated somewhat with simRS (0.45), but more strongly with simPS (0.69). For those with an intermediate simRS, high TOP2A expression (above the median) was associated with significantly higher relapse rates at 5 years (HR 1.82, p = 0.007). TOP2A expression provides prognostic information in patients with ER-positive, HER2-negative breast cancer, a population known to have low incidence of TOP2A gene alterations. These findings confirm prior reports indicating that TOP2A expression provides prognostic information in ER-positive breast cancer. TOP2A expression may also be useful for identifying those with an intermediate RS who are more likely to relapse, although additional validation in datasets including measured rather than simulated RS will be required.     

HER-2 and TOP2A coamplification in urinary bladder cancer

HER-2/NEU is one of the most frequently amplified oncogenes and a potential therapeutic target in bladder cancer. In breast cancer, the adjacent TOP2A gene, the molecular target for several anticancer drugs, is frequently coamplified together with HER-2. To study the amplification and expression of TOP2A and HER-2 and associations with tumor phenotype and clinical outcome in bladder cancer, a tissue microarray containing 2,317 bladder tumor samples was analyzed by FISH and immunohistochemistry. Overall amplification frequencies were 6.3% for HER-2 and 1.5% for TOP2A. Amplifications were most frequently seen in advanced-stage (pT2-4) tumors (HER-2 13.8%, TOP2A 3.4%). Of HER-2-amplified tumors, 56% also had alterations of TOP2A, including 14.7% coamplifications, 33.3% gains and 8% deletions. Only 17.6% of TOP2A amplifications occurred independently of HER-2 alterations. Both HER-2 and TOP2A amplifications were significantly associated with advanced tumor stage (HER-2 p < 0.0001, TOP2A p = 0.0218), high grade (p < 0.0001 for both) and protein overexpression (p < 0.0001 for both). TOP2A amplification and overexpression were linked to shortened survival in muscle-invasive tumors (p = 0.0042 and 0.0077, respectively). In summary, our data suggest that HER-2 amplifications are frequently linked to alterations of the TOP2A gene in bladder cancer. The anatomy of the 17q12-q21 amplicon may be important for response to therapies targeting HER-2 or TOP2A.