代谢综合征、C反应蛋白与冠心病关系的临床研究

目的：探讨代谢综合征（MS）、C反应蛋白与冠心病之间的关系。方法：将230例MS患者分为MS合并冠心病组和MS无冠心病组，比较其体重指数（BMD、甘油三酯（TG）、总胆固醇（TC）、低密度脂蛋自（LDL）、高密度脂蛋白（HDL）、空腹血糖（FBG）、餐后2小时血糖（2hBG）、空腹胰岛素（FINS）、餐后2小时胰岛素（2hINS）、糖化血红蛋白（HbAlc）及胰岛素抵抗指数（ISI）、C反应蛋白（CRP）的变化。结果：MS患者BMI、TG、TC、LDL、FBG、2hBG、FINS、2hINS、HbAlc、CRP较对照组显著升高（P〈0．01），ISI较对照组显著降低（P〈0．01），而MS合并冠心病组较无冠心病组上述改变更显著（P〈0．05）。结论：MS患者存在血脂和血糖代谢紊乱、胰岛素抵抗和炎症标记物CRP升高的病理变化，胰岛素抵抗和炎症状态在MS患者冠心病的发生和发展起着促进作用。     

小窝相关蛋白在乳腺癌中作用的研究进展

摘要：小窝及其相关蛋白与乳腺癌的发生发展密切相关。小窝外壳蛋白包括Cav-1、Cav-2及Cav-3,其中Cav-1最受关注,其作为抑癌及促癌因子影响乳腺癌细胞的增殖、凋亡、迁移、侵袭以及转移。Cav-2也具有抑癌及促癌双重作用,其既可与Cav-1结合共表达,也可独自发挥调控作用。Cav-3在乳腺癌中研究较少,其表达缺失可形成抗肿瘤微环境。小窝接头蛋白（Cavins）包括Cavin-1、Cavin-2、Cavin-3及Cavin-4。Cavin-1可抑制Cav-1诱导的细胞膜小管形成,其在乳腺癌中发挥的具体作用仍存争议。Cavin-2作为乳腺癌抑制因子,可通过阻断转化生长因子（TGF）-β信号通路,抑制乳腺癌进展。Cavin-3在乳腺癌中发挥抑癌功能,而其具体作用机制尚不清楚。Cavin-4与乳腺癌间关系尚不明确。     

乳腺癌组织中ERβ与NF-κB的表达及意义

目的检测乳腺癌组织中雌激素受体（ERβ）和核因子-κB（NF-κB）蛋白的表达,并探讨其意义。方法用免疫组化SP法检测82例乳腺癌组织和45例癌旁正常组织中ERβ和NF-κB蛋白的表达情况。结果①NF-κB蛋白在乳腺癌组织中阳性表达率（45.1%）高于癌旁正常组织（15.6%）（P〈0.05）;ERβ在乳腺癌组织中阳性表达率（25.6%）低于癌旁正常组织（64.5%）（P〈0.05）;②NF-κB在乳腺癌组织中表达随着淋巴结转移增高（P〈0.05）,ERβ的表达与组织分级呈负相关;③NF-κB和ERβ在乳腺癌组织中的表达无关（P〉0.05）。结论 NF-κB在乳腺癌的发生、发展过程中发挥着重要作用,ERβ随细胞恶变发生改变。对NF-κB、ERβ表达水平的联合检测在乳腺癌的诊断、治疗及判断预后方面有一定的指导意义。     

罗格列酮钠对代谢综合征患者糖脂代谢及炎性因子的影响

目的观察罗格列酮钠对代谢综合征（MS）患者胰岛素抵抗和炎性因子影响,探讨其改善MS患者糖脂代谢机制。方法符合诊断的MS患者分为洽疗组56例和正常对照组20例。治疗前后分别测定MS组空腹血糖（FBS）、餐后2h血糖（PBS）、空腹胰岛素（FINS）、C-反应蛋白（CRP）、白介素-6（IL-6）及血脂全套,并用稳态模型计算胰岛素抵抗指数（HOMA—IR）。结果罗格列酮钠治疗后MS患者的血糖、血清胰岛素、HOMA—IR及炎性因子水平明显下降（P〈0．05）;血清三酰甘油（TG）,低密度脂蛋白（LDL—C）和总胆固醇（TCH）水平均明显下降（P〈0．05或P〈0．01）。结论罗格列酮钠改善MS患者胰岛素抵抗的同时还具有抗炎作用,进而改善MS患者的糖脂代谢并能有效防治心血管并发症。     

乳腺癌组织p73蛋白的表达及意义

目的探讨p73蛋自在乳腺癌组织的表达情况及其与乳腺癌生物学行为的关系。方法采用免疫组化Maxvision方法检测63例浸润性乳腺癌患者手术切除标本中癌及癌旁正常组织中p73蛋白的表达情况。结果63例患者乳腺癌p73蛋白的阳性表达率为55。6％,p73的高表达与乳腺癌患者淋巴结转移及雌激素受体（ER）、孕激素受体（PR）、p53的表达有关（P〈0．05）。p73的阳性检出率与乳腺癌患者年龄、肿瘤类型、病理分级等无关（P〉0．05）。结论p73基因在浸润性乳腺癌的发生、发展过程中起着重要的作用,检测p73蛋白可为浸润性乳腺癌的治疗、预后判断提供必要的理论依据。     

生物技术及产品——Pronota公司推出生物标记物平台

比利时Pronota公司已推出其生物标记物验证平台MASSterclass。公司称,这一无抗体系统可减少用于筛选血液中低丰度生物标记物的大量蛋白分析开发的费用和时间。该技术采用了一系列与靶向串联质谱相结合的新的正交复杂性减低步骤（orthogonal complexity reduction）。这一新平台是Pronota公司的生物标记物发现平台的补充,后者可产生一张血液中低丰度蛋白生物标记物的列表。     

环氧化酶-2基因及蛋白在乳腺癌中的表达及意义

目的：探讨环氧化酶-2（cyclooxygenase-2，COX-2）基因及蛋白在乳腺癌中的表达及临床意义。方法：采用原位杂交和S-P免疫组织化学技术检测20例乳腺纤维腺瘤和52例乳腺癌中COX-2mRNA和蛋白的表达。结果：COX-2mRNA和蛋白在乳腺纤维腺瘤和乳腺癌组织中的阳性率分别为（20.0%、65.4%）、（15.0%、57.7%），差异有显著性（P〈0.01）；COX-2mRNA和蛋白表达之间差异无显著性（P〉0.05）。COX-2mRNA表达与淋巴结转移和肿瘤大小有关（P〈0.05），与年龄和组织学分级无关（P〉0.05）；COX-2蛋白表达与淋巴结转移有关（P〈0.05），与年龄、肿瘤大小和组织学分级无关（P〉0.05）。结论：（1）COX-2mRNA及蛋白在乳腺癌中表达上调，提示COX-2在乳腺癌的发生、发展中起重要作用。（2）COX-2与淋巴结转移正相关，提示COX-2可作为判断乳腺癌预后的标记物。     

医用生物蛋白胶对乳腺癌术后皮下积液的预防作用

目的 探讨医用生物蛋白胶对乳腺癌改良根治术后皮下积液的预防效果。方法51例乳腺癌患者分为两组，采用医用生物蛋白胶组25例行乳腺癌改良根治术时在手术创面使用生物蛋白胶，未用医用生物蛋白胶组26例行乳腺癌改良根治术时不使用生物蛋白胶，观察术后第1天引流量、术后总引流量、皮下积液情况，以确定医用生物蛋白胶对乳腺癌改良根治术后皮下积液的预防效果。结果采用医用生物蛋白胶组的第1天引流量和总的引流量分别为（115．04±50．33）ml和（185．20±65．14）ml，与未采用医用生物蛋白胶组的（167．31±77．05）ml和（281．54±106．21）ml比较，差异有统计学意义（P〈0．01）。采用医用生物蛋白胶组患者皮下积液发生率为8．00％（2／25）明显低于未采用医用生物蛋白胶组的34．62％（9／26）（P〈0．05）。结论乳腺癌改良根治术中应用医用生物蛋白胶能有效减少创面引流量，降低皮下积液，起到预防术后皮下积液的作用，有利于切口愈合。     

PCR法检测乳腺癌患者血清抗p53蛋白抗体的研究

目的为乳腺癌诊断提供血清学新方法.方法用免疫-聚合酶链反应（immuno-polymerase chain reanon,PCR）法检测血清抗p53蛋白（一种抑癌基因）抗体,酶免疫组化方法检测组织p53蛋白表达.结果乳腺癌患者血清抗p53蛋白抗体阳性率为39.5%,而非癌患者和正常人血清抗p53蛋白抗体均为阴性,乳腺癌患者血清中抗p53蛋白抗体显著高于非癌患者和正常人（P＜0.01）.p53蛋白阳性表达的乳腺癌患者抗p53蛋白抗体阳性率为64.2%,明显高于p53蛋白阴性表达组,血清p53抗体测定与p53蛋白表达密切相关（P＜0.01）.结论检测血清抗p53蛋白抗体是检测组织p53蛋白理想的替代工具,抗p53蛋白抗体可以作为乳腺癌血清学诊断新标志,用于乳腺癌的普查和早期诊断.     

乳腺癌组织中脆性组氨酸三联体基因的表达及意义

目的探讨乳腺癌组织中脆性组氨酸三联体（FHIT）基因的表达及其临床意义。方法应用免疫组织化学酶联免疫sP法,检测FHIT在41例乳腺癌组织及11例癌旁组织中的表达。结果乳腺癌组FHIT蛋白阳性表达率46．3％（19／41）显著低于正常乳腺组90．9％（10／11）（P〈0．05）。中／高分化组FHIT蛋白阳性表达率显著高于低分化组（53．8％vs33．3％）（P〈0．05）;淋巴结转移组FHIT蛋白阳性率低于,无淋巴结转移组（42．9％vs53．8％）（P〈0．05）;Ⅰ＋Ⅱ期组与Ⅲ＋Ⅳ期组之间FHIT蛋白阳性表达率（56．3％ vs 40．0％）差异有统计学意义（P〈0．05）。结论FHIT基因在乳腺癌的发生、发展中起重要作用,对其蛋白的检测可作为判定乳腺癌发生及转移能力的一项客观指标。     

EZH2 regulates expression of FOXC1 by mediating H3K27me3 in breast cancers

Triple-negative breast cancer (TNBC) is characterized by low expression of human epidermal growth factor receptor-2 (HER2), estrogen receptor (ER), and progesterone receptor (PR), which is the most aggressive subtype with poor outcome among breast cancers. The underlying mechanisms of TNBC remain unclear and there is a lack of biomarkers. In this study we conducted an in silico assay and found that FOXC1 was highly expressed in ER⁻/PR⁻/HER2⁻ breast cancers, which was confirmed by qRT-PCR, immunohistochemistry, and Western blot analysis. FOXC1 was more highly expressed in TNBCs than the other breast cancers. Kaplan–Meier plotter revealed that expression of FOXC1 was associated with overall survival (OS) of patients with breast cancers. Expression of FOXC1 was reversely associated with level of H3K27me3, which was methylated by EZH2. In MCF-7 and T47D cells, inhibition of EZH2 by DZNeP or GSK343 concentration- and time-dependently increased expression of FOXC1. Finally, we demonstrated that the expression of FOXC1 was associated with resistance of doxorubicin treatment of breast cancer cells. In conclusion, these results suggest that FOXC1 may be a potential biomarker or drug target for TNBCs, and that downregulation of FOXC1 could have therapeutic value in treatment of TNBCs.     

基于液相色谱-质谱技术的乳腺癌转移相关 代谢标志物的筛选

摘要：目的 应用代谢组学技术筛选与乳腺癌转移相关的代谢标志物。方法 收集100例乳腺癌患者和50例 健康志愿者的血清标本,采用高效液相色谱-轨道离子阱质谱联用（HPLC-LTQ Orbitrap XL MS）代谢组学研究平台分 析乳腺癌未转移患者、乳腺癌转移患者和健康人群血清标本的代谢轮廓,并通过模式识别方法结合非参数检验对数 据进行分析。结果 由乳腺癌未转移组、乳腺癌转移组和健康对照组的代谢轮廓构建的正交偏最小二乘判别分析 （OPLS-DA）模型具有很好的判别能力（R2=95.2%,Q2=86.7%）,可以鉴别出用于区分乳腺癌转移与否的8个代谢标志 物,包括溶血磷脂酸［18∶1（9Z）/0∶0］、溶血磷脂酰胆碱（18∶0）、溶血磷脂酰胆碱［20∶3（5Z,8Z,11Z）］、胆碱、磷酸二羟 丙酮（18∶0e）、2R,3S-番石榴酸、芥酸、L-氢化乳清酸。结论 利用代谢组学方法获得的血清代谢轮廓可以用来构建 区分模型和寻找乳腺癌转移相关的代谢标志物,为乳腺癌的早期诊治、预后评估和药物治疗靶点的选择提供支持和 依据。     

应用SELDI—TOFMS技术筛选乳腺癌蛋白标志物

目的：应用表面增强激光解吸电离飞行时间质谱技术（ SELDI-TOF MS）筛选乳腺癌的特异性蛋白标志物,建立诊断模型。方法用表面增强激光解吸电离飞行时间质谱仪及CM10蛋白芯片检测35例乳腺癌患者标本及53例对照组标本（包括35例乳腺良性病变和18例正常人）的血清蛋白指纹图谱,Ciphergen Proteinchip 软件自动采集数据,Ciphergen Biomaker Wizard 软件筛选差异蛋白,Biomarker Pattern软件建立乳腺癌的分类树诊断模型。结果乳腺癌组及对照组血清蛋白质谱图共检测到59个蛋白质峰,其中19个蛋白峰表达差异具有显著性（P＜0．01）。以2个蛋白质峰（质荷比分别为M6636．62,M13889．6）建立的诊断模型,诊断的准确率达到94．3％,灵敏度和特异度分别为80．0％和71．7％。结论 SELDI-TOF MS技术可用于乳腺癌特异性蛋白的筛选,为其快速诊断奠定基础。     

Investigation of the Ovarian and Prostate Cancer Peptidome for Candidate Early Detection Markers Using a Novel Nanoparticle Biomarker Capture Technology

Current efforts to identify protein biomarkers of disease use mainly mass spectrometry (MS) to analyze tissue and blood specimens. The low-molecular-weight “peptidome” is an attractive information archive because of the facile nature by which the low-molecular-weight information freely crosses the endothelial cell barrier of the vasculature, which provides opportunity to measure disease microenvironment-associated protein analytes secreted or shed into the extracellular interstitium and from there into the circulation. However, identifying useful protein biomarkers (peptidomic or not) which could be useful to detect early detection/monitoring of disease, toxicity, doping, or drug abuse has been severely hampered because even the most sophisticated, high-resolution MS technologies have lower sensitivities than those of the immunoassays technologies now routinely used in clinical practice. Identification of novel low abundance biomarkers that are indicative of early-stage events that likely exist in the sub-nanogram per milliliter concentration range of known markers, such as prostate-specific antigen, cannot be readily detected by current MS technologies. We have developed a new nanoparticle technology that can, in one step, capture, concentrate, and separate the peptidome from high-abundance blood proteins. Herein, we describe an initial pilot study whereby the peptidome content of ovarian and prostate cancer patients is investigated with this method. Differentially abundant candidate peptidome biomarkers that appear to be specific for early-stage ovarian and prostate cancer have been identified and reveal the potential utility for this new methodology     

代谢组学在不同分子亚型乳腺癌中的研究进展

乳腺癌的治疗和预后受到分子分型影响,明确分子分型可让乳腺癌诊治更精准。代谢组学是继基因组学、转录组学和蛋白质组学后发展起来的一种组学技术,被广泛应用于肿瘤标志物的筛选和疾病诊断。近年来研究发现,乳腺癌各亚型之间具有代谢重编程的异质性,代谢组学所挖掘出的生物标志物为乳腺癌分子亚型的早期诊断、预后评估、药物靶点和机制研究以及毒理研究提供了高特异性、高灵敏度和高选择性的可视化指标。本文针对可作为乳腺癌各分子亚型的代谢标志物进行综述,以期为不同亚型乳腺癌的诊断和精准化治疗提供了一种新思路。     

Therapies for triple negative breast cancer

Introduction: Triple negative breast cancer (TNBC) is a heterogeneous disease associated with a high risk of recurrence, and therapeutic options are currently limited to cytotoxic therapy. Germ-line mutations may occur in up to 20% of unselected patients with TNBC, which may serve as a biomarker identifying which patients may have tumors that are particularly sensitive to platinums and/or inhibitors of poly(ADP-ribose)polymerase. A substantial proportion of patients with TNBCs not associated with germ-line BRCA mutations may have tumors that are ‘BRCA-like’, rendering those individuals potential candidates for similar strategies.

Areas covered: The purpose of this review is to highlight the current standard and experimental treatment strategies.

Expert opinion: Recent research that has illuminated the molecular heterogeneity of the disease rationalizes its diverse biological behavior and differential response to chemotherapy. Modern technology platforms provide molecular signatures that can be mined for therapeatic interventions. Target pathways that are commonly dysregulated in cancer cells control cellular processes such as apoptosis, proliferation, angiogenesis, DNA repair, cell cycle progression, immune modulation and invasion, and metastasis. Novel trial design and re-defined endpoints as surrogates to clinical outcome have been introduced to expedite the development of breakthrough therapies to treat high-risk early-stage breast cancer.     

cMET in triple-negative breast cancer: is it a therapeutic target for this subset of breast cancer patients?

Introduction: The identification and validation of a targeted therapy for triple-negative breast cancer (TNBC) is currently one of the most urgent needs in breast cancer therapeutics. The cMET oncogene encodes a membrane-bound tyrosine kinase implicated in the formation and/or progression of several cancer types, including TNBC. Currently, inhibitors targeting cMET are undergoing clinical trials for a variety of cancers, including TNBC. These include anti-cMET and anti-hepatocyte growth factor (HGF) monoclonal antibodies and tyrosine kinase inhibitors.

Areas covered: This article reviews the structure and mode of action of cMET, the role of cMET in cancer formation/development, with particular emphasis on its role in basal/TNBC and its potential as a therapeutic target for this subtype of breast cancer.

Expert opinion: Due to cancer heterogeneity, it is unlikely that all TNBC patients will be responsive to anti-cMET drugs. Therefore, if cMET is to be used as a target for treatment, it will be important to identify predictive biomarkers to select, upfront, those patients likely to benefit. Potential predictive biomarkers for anti-cMET treatments in basal/TNBC include cMET, phospho-cMET, downstream signaling proteins or HGF. These putative predictive biomarkers should be evaluated in a large panel of basal/TNBC cell lines before incorporation into clinical trials involving anti-cMET drugs.     

Lemur tyrosine kinase-3 (LMTK3) as a target in oestrogen positive breast cancer

Oestrogen receptor positive (ER⁺) breast cancers are susceptible to endocrine treatments, such as tamoxifen. However, resistance is a major problem when treating breast cancer with these agents. ERα phosphorylation is implicated in this resistance to tamoxifen, and kinases are the enzymes that catalyse phosphorylation. A kinome search in human breast cancer cells identified lemur tyrosine kinase 3 (LMTK3). In primary breast cancer samples, high nuclear LMTK3 expression was associated with a shorter disease-free survival time. In tamoxifen-resistant cell lines, the addition of LMTK3 siRNA, increased the inhibitory effect of tamoxifen. In nude mice, with established human MCF-7 breast cancer tumours, LMTK3 siRNA decreased tumour growth. In conclusion, LMKT3 is a possible target and marker of breast cancer.     

肝素结合细胞因子影响乳腺癌细胞的增殖、侵袭和上皮细胞间质化的功能研究

目的 探讨肝素结合细胞因子(Midkine)的表达对乳腺癌细胞的增殖和侵袭的作用.方法 采用慢病毒介导的shRNA干扰技术在乳腺癌细胞中沉默Midkine的表达,及利用慢病毒高表达Midkine基因在正常乳腺表皮细胞中.利用蛋白质印迹(Western blot)法检测shRNA干扰或高表达后的正常乳腺表皮细胞和乳腺癌细胞中Mid-kine蛋白的表达情况.利用流式细胞仪技术、Transwell小室实验、及细胞划痕实验,分析Midkine基因高低表达对正常乳腺表皮细胞和乳腺癌细胞生长、增殖、侵袭及转移的影响.结果 首先观察了不同乳腺癌细胞中Midkine的表达情况.其中Midkine只有在BT549、MDA-MB157和MDA-MB436间充质乳腺癌细胞(Mesenchymal cell)中表达,而正常细胞和上皮细胞乳腺癌细胞(Epithelial cell)中不表达.BT549细胞中两种不同shRNA降低Mdikine蛋白表达的效率分别达到100%和90%.HMLE正常乳腺上皮细胞中Midkine高表达效率非常高.结晶紫染色法检测细胞增殖实验结果显示,Midkine-shRNAs转染组BT549细胞的生长速度较阴性对照组降低(P<0.01).流式细胞仪检测结果显示,Midkine-shRNAs转染组BT549细胞的增殖指数为(36.06±2.12)%和(32.06±3.46)%,明显低于对照组53.06±3.68%.同时观察到EMT标识物B连环蛋白(β-catenin)、N钙粘蛋白(N-cadherin)在Midkine-shRNAs转染组BT549细胞中表达上调.Transwell小室实验显示两个Midkine-shRNA转染组BT549细胞迁移(Migration)细胞数为零和(7±4.4)个高/倍视野,它们的侵袭(Invasion)细胞数为(38±7)和(46±8)个/高倍视野,明显低于对照组的迁移(Migra-tion)细胞数(178±14)个/高倍视野,侵袭(Invasion)细胞数(232±35)个/高倍视野.同时也观察到高表达Midkine正常乳腺上皮细胞表现出间质化(EMT)的表型,EMT标识物E钙粘蛋白(E-cadherin)、连环蛋白(β-catenin)在Midkine高表达HMLE细胞中表达下调的现象.细胞划痕实验显示Midkine高表达HMLE细胞组的迁移距离明显高于对照组.结论 在BT549细胞中降低Midkine的表达,可抑制BT549细胞的迁移和侵袭,同时可明显上调对上皮细胞间质化标志蛋白β-catenin和N-cadherin表达,提示在Midkine高表达的肿瘤患者中,通过抑制其蛋白表达可能抑制肿瘤的转移和侵袭.     

Biomarkers in breast cancer

Breast cancer is one of the most frequently diagnosed cancers among women in the western world. Due to the aggressive behaviour of some specific types and the possibility of an early diagnosis, breast cancer has been constantly studied. Tumour size, histological type, cellular and nuclear characteristics, mitotic index, vascular invasion, hormonal receptors and axillary lymph node status are biomarkers routinely used. However, these parameters are not enough to predict the course of this disease. Molecular biology advances have made it possible to find new markers, which have already been incorporated to the clinical practice. Their ultimate goal is to reduce mortality by identifying women at risk for the development of this disease, help diagnosis, determine prognosis, detect recurrences, monitor and guide treatment, and in particular cancers they are suited for general screening. Tumour markers in breast cancer were ranked in categories reflecting their clinical utility, according to the American College of Pathologists. This article focuses on traditional and new molecular markers stratifying them into categories and emphasizing their relevance in the routine evaluation of patients with breast cancer.