bcl—2基因在乳腺癌中的表达及临床意义

目的研究bcl-2基因在乳腺癌癌灶、癌旁组织、转移淋巴结及正常组织中的表达,与肿瘤形成、发展的关系及临床意义.方法应用相对定量RT-PCR方法,检测bcl-2基因在16例乳腺癌癌灶、癌旁组织、转移淋巴结及正常乳腺组织中的表达.结果乳腺癌组织中bcl-2 mRNA表达相对值(0.264±0.137),明显高于正常乳腺组织表达相对值(0.033±0.031)(P<0.005);转移淋巴结bcl-2 mRNA表达相对值(0.272±0.076),癌旁组织bcl-2mRNA表达相对值(0.131±0.083),明显高于正常乳腺组织表达相对值(P<0.05);淋巴结转移之癌灶组bcl-2 mRNA表达相对值(0.381±0.096)明显高于非转移之癌灶组(0.186±0.102)(P<0.05).结论bcl-2基因的高表达可能在乳腺癌的发生、发展中起一定的作用.     

Overexpression of the pp32r1 (ANP32C) oncogene or its functional mutant pp32r1Y140H confers enhanced resistance to FTY720 (Finguimod)

pp32r1 (ANP32C) is oncogenic and has been shown to be overexpressed in tumors of the breast, prostate, and pancreas. In this work we show that pp32 family proteins are able to bind to the sphingosine analog FTY720 (Finguimod). Molecular docking studies highlight that a conserved residue F136 is likely to be a key determinant of the FTY720 binding site on the pp32 leucine-rich repeat domain. Transduction of the renal carcinoma cell line ACHN or cervical cancer cell line HeLa with lentivirus expressing the oncogenic family member pp32r1 or a pp32r1Y140H functional mutant illustrated an enhanced resistance to FTY720 induced apoptosis. These findings highlight that certain cancers overexpressing pp32r1 or pp32r1 mutants are likely to demonstrate enhanced resistance to FTY720 treatment.     

乳腺增生病p16蛋白表达及p16基因突变

目的　探讨p16基因在乳腺癌发生早期的作用。方法　用免疫组化方法检测 36例乳腺单纯性增生、31例不典型增生、14例导管内癌和 16例浸润性导管癌中p16蛋白的表达 ,用PCR SSCP和DNA测序技术检测上述组织中p16基因突变。结果　p16蛋白在单纯性增生、不典型增生、导管内癌、浸润癌中的表达缺失率分别为 0、16 .1%( 5 /31)、35 .7%( 5 /14 )、43.7%( 7/16 ) ,在 1例不典型增生、1例导管内癌、1例浸润性导管癌中检测p16基因突变。结论　p16蛋白表达缺失出现在乳腺癌发生的早期阶段 ,这种表达异常似乎与p16基因突变无明显关系。     

Dynamic classification using case‐specific training cohorts outperforms static gene expression signatures in breast cancer

The molecular diversity of breast cancer makes it impossible to identify prognostic markers that are applicable to all breast cancers. To overcome limitations of previous multigene prognostic classifiers, we propose a new dynamic predictor: instead of using a single universal training cohort and an identical list of informative genes to predict the prognosis of new cases, a case‐specific predictor is developed for each test case. Gene expression data from 3,534 breast cancers with clinical annotation including relapse‐free survival is analyzed. For each test case, we select a case‐specific training subset including only molecularly similar cases and a case‐specific predictor is generated. This method yields different training sets and different predictors for each new patient. The model performance was assessed in leave‐one‐out validation and also in 325 independent cases. Prognostic discrimination was high for all cases (n = 3,534, HR = 3.68, p = 1.67 E?56). The dynamic predictor showed higher overall accuracy (0.68) than genomic surrogates for Oncotype DX (0.64), Genomic Grade Index (0.61) or MammaPrint (0.47). The dynamic predictor was also effective in triple‐negative cancers (n = 427, HR = 3.08, p = 0.0093) where the above classifiers all failed. Validation in independent patients yielded similar classification power (HR = 3.57). The dynamic classifier is available online at http://www.recurrenceonline.com/?q=Re_training . In summary, we developed a new method to make personalized prognostic prediction using case‐specific training cohorts. The dynamic predictors outperform static models developed from single historical training cohorts and they also predict well in triple‐negative cancers.     

黑色素瘤相关抗原A基因在乳腺癌中的表达及其临床意义

目的: 探讨黑色素瘤相关抗原A家族(MAGE-As)在乳腺癌患者外周血及组织中的表达,并分析其与乳腺癌患者临床病理学指标及预后的关系。方法: 采用多重巢式PCR法检测106例乳腺癌患者和30例健康志愿者外周血中MAGE-As m RNA的表达水平,并利用限制性内切酶酶切法,鉴定该家族成员MAGE-A1、A2、A3、A4和A6的表达情况。采用免疫组织化学法检测了106例乳腺癌组织蜡块中MAGE-As蛋白的表达情况,同时分析外周血中MAGE-As mRNA表达与其相应肿瘤组织中MAGE-As蛋白的表达的相关性。结果: MAGE-As mRNA在乳腺癌患者外周血中阳性表达率为40.5%(43/106)。MAGE-A1、A2、A3、A4和A6的表达率分别为10.3%(11/106)、19.8%(21/106)、15.1%(16/106)、13.2%(14/106)和5.7%(6/106),MAGE-As表达率的顺序为A2>A3>A4>A1>A6。MAGE-As mRNA阳性表达与患者年龄、肿瘤大小、临床分期、淋巴结转移相关(P<0.05)。乳腺癌患者外周血中MAGE-As mRNA表达水平与其相应肿瘤组织中蛋白水平的表达呈正相关(r=0.368,P<0.01)。MAGE-As各亚型(MAGE-A1、A2、A3、A4和A6)阳性表达率与乳腺癌患者5年生存率相关(P<0.01);MAGE-As表达及淋巴结转移可作为乳腺癌患者5年生存率的独立预后影响因素(均P<0.01)。结论: MAGE-As可能作为检测乳腺癌患者外周血中循环肿瘤细胞的特异性生物标志物,其表达与乳腺癌患者预后相关。乳腺癌患者外周血中MAGE-As检测可作为监测乳腺癌预后的重要指标。     

家族性乳腺癌家系成员乳腺癌易感基因突变的研究

目的研究河北省地区家族性乳腺癌家系中的患者及健康一级亲属乳腺癌易感基因1（BRCAl）和乳腺癌易感基因2（BRCA2）突变位点及携带情况。方法研究对象为2002年6月至2008年5月河北医科大学第四医院接诊的乳腺癌患者及其亲属,分别来自12个独立的汉族家族性乳腺癌家系,该家系中有2个及2个以上一级或二级亲属乳腺癌患病史,研究病例包括13例患者及46例健康一级亲属,共59例样本。由外周血提取基因组DNA,采用聚合酶链反应一单链构象多态性分析（PCR—SSCP）和基因测序技术对国内外报告中常见的4个BRCAl／BRCA2突变热点区域（BRCAl：外显子2、11、20;BRCA2外显子11）进行检测。结果发现1个BRCAl突变位点（4193insA）和1个BRCA2突变位点（5329insT）,全部为移码突变;发现4个变异位点（BRCAl：4165T〉A、287G〉C,BRCA2：6251G〉T、5416C〉A）,4193insA、5329insT、287G〉C携带者的家系中均有3例乳腺癌患者。结论BRCAl（4193insA）、BRCA2（5329insT）以及BRCAl：4165T〉A、287G〉C和BRCA2：6251G〉T、5416C〉A可能是河北省家族性乳腺癌相关性突变位点,其携带者家系中乳腺癌发病率明显升高,建议对其一级亲属密切随访或尽早进行手术或药物干预。     

同源异型盒基因HOXD10在乳腺癌组织中的表达及其临床病理意义

背景与目的：乳腺癌是威胁女性健康最常见的恶性肿瘤之一,从分子水平深入研究其癌变机制,对寻找新的标志基因具有重要意义.本研究探讨同源异型盒基因HOXD10基因在乳腺癌发生、发展进程中的可能作用.方法：以18s rRNA基因作为参照,采用荧光实时定量RT-PCR技术,以相对定量方法检测44例乳腺癌组织及16例乳腺良性病变组织中HOXD10 mRNA表达情况.结果：HOXD10在乳腺癌中的表达显著低于乳腺良性病变组织.组织学分级为Ⅲ级的乳腺癌标本中H0XD10 mRNA的表达显著低于组织学Ⅰ、Ⅱ级标本.H0XD10 mRNA的表达与淋巴结转移、肿瘤大小、TNM分期之间相关性不显著.结论：HOXD10基因在乳腺癌中呈低表达甚至表达缺失,由于这种基因可能为乳腺癌发生发展的抑制因素,因此对其我们应给予更多关注.     

Expression of glyoxalase I and II in normal and breast cancer tissues

The present work aimed to study the activities of glyoxalase system enzymes, glyoxalase I (G I) and glyoxalase II (G II), as well as the expression of their genes in human breast carcinoma. Samples of tumoral tissue and normal counterparts were drawn from several patients during surgery. They served either for preparing extracts to be used in enzyme activity evaluations or for RNA extraction and subsequent northern blot analysis. A far higher activity level of G I and G II occurs in the tumor compared with pair-matched normal tissue, as shown by both spectrophotometrical assay and electrophoretic pattern. Such increased activities of G I and G II likely result from an enhanced enzyme synthesis as a consequence of increased expression of the respective genes in the tumoral tissue, as evidenced by northern blot. The present findings confirm a key-role of glyoxalase system to detoxify cytotoxic methylglyoxal and modulate S-D-lactoylglutathione levels in tumor cells. Moreover, they suggest a possible employment of GI inhibitors as anti-cancer drugs.     

乳腺癌耐药蛋白的研究进展

乳腺癌耐药蛋白(BVRP)是新近发现的一种肿瘤耐药相关蛋白．与P-gp和多药耐药相关蛋白(MRP)同属ABC转运蛋白超家族。本文对BCRP的发现、基因表达特征、与造血干细胞分化的关系、转运底物及其耐药逆转和临床意义等方面的研究进展进行综述。     

BRCA1基因启动子甲基化在三阴性乳腺癌中的表达及其临床意义

目的:评估三阴性乳腺癌（TNBC）患者中乳腺癌易感基因1（BRCA1）甲基化的表达及与患者预后的关系。方法:收集在我院甲乳科治疗的乳腺癌患者手术切除标本524例,应用联合亚硫酸氢钠限制性内切酶分析法,观察BRCA1启动子甲基化状态。应用定量逆转录聚合酶链反应评估BRCA1 mRNA表达,应用免疫组织化学法评估BRCA1蛋白表达。结果:共有157（30.0%）例TNBC患者,有25（4.77%）例存在BRCA1启动子甲基化,所有BRCA1启动子甲基化的肿瘤均为TNBC。TNBC患者中,BRCA1启动子甲基化患者的BRCA1 mRNA水平显著低于BRCA1启动子未甲基化患者［（0.019±0.005） vs （0.095±0.013）,P<0.001］,免疫组化分析未在BRCA1启动子甲基化患者中检测出BRCA1蛋白表达。BRCA1启动子甲基化患者的总生存率（OS）显著低于非甲基化患者（logrank P=0.038）。BRCA1蛋白低表达患者的OS与RFS均低于高表达组,但差异没有统计学意义（分别logrank P=0.526,P=0.467）。结论:BRCA1启动子甲基化导致了BRCA1表达的下降,并与TNBC患者较差预后相关。BRCA1启动子甲基化是一种促成BRCA1功能丧失的重要机制。     

Systematic characterisation of GABRP expression in sporadic breast cancer and normal breast tissue

The GABRP gene has been previously identified by in silico analysis of four million ESTs as a candidate gene differentially expressed in breast cancer. GABRP is located on chromosome 5q34 and it encodes the pi-subunit of the gamma-aminobutyric acid (GABA) receptor, a transmembrane protein expressed in the brain and several nonneuronal tissues. Using cDNA dot blot hybridisation (cancer profiling array), quantitative RT-PCR and non-radioisotopic in situ hybridisation (ISH), we have analysed GABRP expression in breast cancer and normal breast tissues as well as in nontumorigenic and tumorigenic breast cell lines. Analysis of the cancer profiling array revealed a more than 2-fold downregulation of GABRP (p < 0.001) in 76% of primary breast carcinomas (n = 50) compared to corresponding normal tissues. Quantitative RT-PCR in a panel of 23 normal human tissues showed that the GABRP expression level was most abundant in the normal breast tissues compared to other human tissues. GABRP downregulation in breast cancer was confirmed by quantitative RT-PCR in cryopreserved breast tumour and normal breast tissue specimens (n = 22), in archival formalin-fixed, paraffin-embedded tissue specimens (n = 32), as well as in breast cancer cell lines (n = 8). Furthermore, a significant downregulation of GABRP was noted in large (pT3-pT4) (p = 0.044) primary breast tumours. Non-radioisotopic ISH showed strong GABRP expression in normal epithelial and benign papilloma breast cells, but no signal could be detected in invasive ductal carcinoma. Altogether, these data suggest that GABRP is progressively down-regulated with tumour-progression, and that it may be useful as a prognostic marker in breast cancer.     

乳腺癌组织BAG-1表达及其与临床病理因素相关性的分析

目的:进一步了解BAG-1在乳腺癌组织中的表达及其与乳腺癌临床病理因素的相关性。方法:利用组织芯片技术以及DAKOEnVision免疫组织化学方法对乳腺癌组织及其对应的癌旁组织和正常组织中BAG-1进行检测,同时分析BAG-1与乳腺癌临床病理因素的相关性。结果:BAG-1经免疫组化DA-KOEnVision染色呈棕黄色颗粒状,BAG-1表达主要定位于阳性细胞的胞质和胞核中。BAG-1在癌组织中的阳性表达率高于癌旁组织和正常组织,P均<0.05。BAG-1的表达与乳腺癌肿块大小、分级及腋窝淋巴结转移无相关性;但与乳腺癌的ER、PR、5年生存率表达呈正相关,与HER-2表达呈负相关,P均<0.01。结论:检测乳腺癌组织中BAG-1的表达,对评估乳腺癌的临床特性及预后有一定的意义。     

The gene expression signature of genomic instability in breast cancer is an independent predictor of clinical outcome

Recently, expression profiling of breast carcinomas has revealed gene signatures that predict clinical outcome, and discerned prognostically relevant breast cancer subtypes. Measurement of the degree of genomic instability provides a very similar stratification of prognostic groups. We therefore hypothesized that these features are linked. We used gene expression profiling of 48 breast cancer specimens that profoundly differed in their degree of genomic instability and identified a set of 12 genes that defines the 2 groups. The biological and prognostic significance of this gene set was established through survival prediction in published datasets from patients with breast cancer. Of note, the gene expression signatures that define specific prognostic subtypes in other breast cancer datasets, such as luminal A and B, basal, normal‐like, and ERBB2+, and prognostic signatures including MammaPrint® and Oncotype DX, predicted genomic instability in our samples. This remarkable congruence suggests a biological interdependence of poor‐prognosis gene signatures, breast cancer subtypes, genomic instability, and clinical outcome. © 2008 Wiley‐Liss, Inc.     

Mutational analysis of the MTHFR gene in breast cancer patients of Pakistani population

Objectives: Since methylenetetrahydrofolatereductase (MTHFR) maintains the balance of circulating folate and methionine and blocks the formation of homocysteine, its regulation in relation to different cancers has extensively been studied in different populations. However, information on Pakistani breast cancer patients is lacking. The MTHFR gene has two most common mutations that are single nucleotide additions which result in change of amino acids C677T to Ala222val and A1298C to Glu429Ala. Methodology: 110 sporadic breast patients with no prior family history of cancer or any other type of genetic disorders along with 110 normal individuals were screened for mutations in exons 1 to exon 9 using single strand conformational polymorphism, RFLP and sequencing analyzer. Results: The p values for the 677CC, 677CT, and 677TT genotypes were 0.223, 0.006, and 0.077, respectively. Those for the 1298AA, 1298AC, and 1298CC genotypes were 0.555, 0.009, and 0.003, respectively. Conclusions: We found an overall a significant, weak inverse association between breast cancer risk and the 677TT genotype and an inverse association with the 1298C variant. These results for MTHFR polymorphism might be population specific in sporadic breast cancer affected patients but many other factors need to be excluded before making final conclusions including folate intake, population and disease heterogeneity.