本文關(guān)鍵詞：中國(guó)乳腺癌HER2基因檢測(cè)、臨床病理特征分析及流行病學(xué)研究，由筆耕文化傳播整理發(fā)布。

        目前,分子診斷技術(shù)在乳腺癌個(gè)體化診療的臨床實(shí)踐中發(fā)揮著越來越重要的作用。依據(jù)受體狀態(tài)選擇乳腺癌內(nèi)分泌治療和分子靶向治療成為個(gè)體化診療的典范,尤其是人表皮生長(zhǎng)因子受體2(HER2)在乳腺癌治療中的地位越來越受重視,不僅可以預(yù)測(cè)細(xì)胞毒性藥物蒽環(huán)類或紫杉類藥物的療效,而且已經(jīng)成為評(píng)價(jià)抗HER2靶向藥物臨床獲益的重要分子標(biāo)志物。針對(duì)HER2的單克隆抗體曲妥珠單抗,使HER2陽(yáng)性乳腺癌患者復(fù)發(fā)風(fēng)險(xiǎn)明顯降低,無論單藥、與化療聯(lián)合或輔助治療均能夠持續(xù)改善患者的無病生存時(shí)間,因其在乳腺癌治療取得的卓越療效,成為腫瘤分子靶向治療的代表。因此,根據(jù)HER2表達(dá)狀態(tài)制定細(xì)胞毒性藥物及靶向藥物的合理治療方案,并預(yù)測(cè)藥物療效和評(píng)價(jià)預(yù)后已經(jīng)成為乳腺癌診療的主要策略。那么,如何準(zhǔn)確檢測(cè)HER2的狀態(tài),為乳腺癌患者的診斷、治療和評(píng)價(jià)提供依據(jù),成為目前一項(xiàng)意義重大的緊迫任務(wù)。HER2定位于17q12,是表皮生長(zhǎng)因子受體家族(EGFR)的成員之一,HER2基因可以編碼分子量為185KDa的酪氨酸激酶活性跨膜糖蛋白。HER2與配體結(jié)合形成二聚體,構(gòu)象發(fā)生改變,導(dǎo)致細(xì)胞內(nèi)受體酪氨酸激酶的磷酸化,引起信號(hào)通路上的連鎖反應(yīng),促進(jìn)細(xì)胞增殖分化。國(guó)內(nèi)外研究報(bào)道,HER2過度表達(dá)與乳腺癌的發(fā)生、發(fā)展、侵襲和轉(zhuǎn)移密切相關(guān)。然而,隨著國(guó)內(nèi)HER2檢測(cè)的逐步開展以及曲妥珠單抗的臨床應(yīng)用,在HER2檢測(cè)準(zhǔn)確性及其臨床實(shí)踐中的指導(dǎo)意義方面暴露出一系列問題：一、HER2的檢測(cè)方法多種多樣,但檢測(cè)技術(shù)的標(biāo)準(zhǔn)化、檢測(cè)結(jié)果的準(zhǔn)確性、不同方法的一致性以及進(jìn)口檢測(cè)試劑價(jià)格昂貴等問題亟待解決。二、中國(guó)乳腺癌患者HER2表達(dá)的流行病學(xué)情況是否與國(guó)外存在差異仍不明確。三、17號(hào)染色體多倍體在中國(guó)乳腺癌患者中的表達(dá)情況,對(duì)HER2狀態(tài)的影響,以及與臨床病理特征的關(guān)系仍不清楚。四、臨床實(shí)踐中是否可以用血清代替腫瘤組織標(biāo)本檢測(cè)HER2的狀態(tài)仍存在爭(zhēng)議。為此,我們?cè)噲D從以上幾個(gè)方面進(jìn)行研究,研究?jī)?nèi)容分為四個(gè)部分：第一部分：國(guó)產(chǎn)GLP和進(jìn)口PathVysion FISH探針試劑盒檢測(cè)乳腺癌HER2基因狀態(tài)的比較研究。通過收集108例乳腺浸潤(rùn)性導(dǎo)管癌腫瘤組織標(biāo)本,分別采用國(guó)產(chǎn)試劑盒GLP和進(jìn)口試劑盒Vysis PathVysion探針檢測(cè)乳腺癌患者腫瘤組織HER2基因表達(dá)水平,比較2種試劑檢測(cè)HER2基因狀態(tài)的差異。結(jié)果表明,與Vysis探針相比,國(guó)產(chǎn)GLP HER2探針試劑盒在檢測(cè)乳腺癌HER2基因擴(kuò)增的敏感度、特異度、陽(yáng)性預(yù)測(cè)值和陰性預(yù)測(cè)值均高,且具有價(jià)格優(yōu)勢(shì),在國(guó)內(nèi)臨床評(píng)價(jià)乳腺癌HER2基因狀態(tài)中具有廣泛應(yīng)用價(jià)值。第二部分：中國(guó)乳腺癌HER2表達(dá)的流行病學(xué)研究。運(yùn)用免疫組織化學(xué)(IHC)和原位雜交技術(shù)(FISH)分別評(píng)價(jià)了不同地域和不同種族3,149例中國(guó)乳腺癌患者HER2的表達(dá)水平,分析了HER2的表達(dá)與地理分布及臨床病理特征的相關(guān)性。結(jié)果表明,中國(guó)乳腺癌患者發(fā)病年齡較西方早,臨床病理特征與美國(guó)和歐洲相比,腫瘤負(fù)荷較大(p<0.001)、組織分級(jí)較差(p<0.001)、且ER/PR多為陰性(p<0.001)。提示,中國(guó)乳腺癌患者的腫瘤侵襲性可能更強(qiáng)。HER2蛋白過表達(dá)率和基因擴(kuò)增率分別為23.3%和27.5%,兩者一致率達(dá)71.2%(Kappa=0.494,p<0.001)。HER2基因擴(kuò)增與臨床分期晚(Ⅲ/Ⅳ)(p=0.002)、腫瘤負(fù)荷大(>5cm)(p=0.002)、組織分級(jí)差(p<0.001)、絕經(jīng)(p<0.001)、ER/PR表達(dá)陰性(p=0.002)以及淋巴結(jié)轉(zhuǎn)移≥4個(gè)(p<0.001)密切相關(guān)。南方與北方乳腺癌患者的HER2擴(kuò)增狀態(tài)無顯著差異(p>0.05)。本研究揭示了中國(guó)乳腺癌HER2蛋白過表達(dá)和基因擴(kuò)增流行病學(xué)特征,以及病理特征與西方人群的差異,為乳腺癌個(gè)體化治療和早期預(yù)防提供有價(jià)值的理論依據(jù)。第三部分：17號(hào)染色體多倍體與HER2表達(dá)的相關(guān)性研究。運(yùn)用FISH技術(shù)檢測(cè)了348例乳腺癌患者的HER2基因和17號(hào)染色體CEP17拷貝數(shù)的情況,結(jié)合HER2蛋白表達(dá)(IHC)情況和相關(guān)臨床病理特征進(jìn)行分組對(duì)比分析。結(jié)果表明,所有患者中CEP17平均拷貝數(shù)為2.1(1.0-12.4),17號(hào)染色體多倍體的比率(CEP17≥3)為13.8%(48/384)。HER2基因拷貝數(shù)>6的患者占26.4%(92/348),HER2/CEP17>2.2的占25.3%(88/348)。HER2拷貝數(shù)>6的患者中,17號(hào)染色體多倍體的比率占38.0%(35/92),HER2蛋白表達(dá)與HER2基因拷貝數(shù)以及比值密切相關(guān)。17號(hào)染色體多倍體與HER2蛋白表達(dá)或基因擴(kuò)增相關(guān),但相關(guān)性較差(分別為r=0.271,p<0.05和r=0.223,p<0.05)。在HER2拷貝數(shù)≤6的患者中,17號(hào)染色多倍體的比率占5.4%(5/92)。單純17號(hào)染色多倍體組的臨床病理特征更傾向于HER2陰性組的病理特征。提示,17號(hào)染色體信號(hào)的增加影響了部分HER2結(jié)果的判讀,但單純的17號(hào)染色體多倍體并不能作為HER2基因擴(kuò)增的獨(dú)立預(yù)測(cè)因子。第四部分：檢測(cè)乳腺癌患者血清中HER2ECD的水平。本研究收集了我院Ⅰ-ⅢA期乳腺癌患者術(shù)前血232例,運(yùn)用ELISA法檢測(cè)血清HER2ECD水平,IHC法和FISH法檢測(cè)配對(duì)腫瘤組織HER2的表達(dá),分析血清HER2ECD表達(dá)與臨床病理特征關(guān)系及臨床意義。結(jié)果表明,血清HER2ECD的中位濃度為6.8ng/ml,HER2ECD水平升高的界值為7.4ng/ml,低于轉(zhuǎn)移性乳腺癌血清HER2ECD水平的臨界值(15ng/ml)。89例(38.3%)患者血清HER2ECD水平升高,而腫瘤組織77例(33.2%)HER2陽(yáng)性表達(dá),一致率達(dá)76.7%。血清HER2ECD升高與絕經(jīng)(p<0.001)、高級(jí)別組織分級(jí)(p<0.001).ER-(p=0.007).PR-(p<0.001).CA153水平升高(p=0.039)和TPS水平升高(p=0.018)密切相關(guān)。與單純檢測(cè)腫瘤組織HER2表達(dá)相比,同時(shí)聯(lián)合檢測(cè)術(shù)前血清中HER2ECD水平能提供更多的信息。我們支持降低術(shù)前血清中HER2ECD的界值指標(biāo)更有利于評(píng)價(jià)乳腺癌患者HER2的狀態(tài)及預(yù)后。本研究通過回答上述問題,明確了HER2國(guó)產(chǎn)試劑盒廣泛的應(yīng)用價(jià)值、獲得了中國(guó)乳腺癌HER2表達(dá)的流行病學(xué)數(shù)據(jù)、了解了17號(hào)染色體多倍體如何影響HER2的表達(dá)、闡述了血清代替組織標(biāo)本檢測(cè)HER2狀態(tài)的可行性,為今后臨床實(shí)踐中準(zhǔn)確判讀HER2狀態(tài)提供了依據(jù),推動(dòng)了HER2檢測(cè)技術(shù)的標(biāo)準(zhǔn)化及乳腺癌個(gè)體化靶向治療的臨床應(yīng)用。目的通過與進(jìn)口Vysis PathVysion HER2探針試劑盒(簡(jiǎn)稱“VysisPathVysion探針試劑”)比較,評(píng)價(jià)國(guó)產(chǎn)金菩嘉GLP HER2探針試劑盒(簡(jiǎn)稱"GLP探針試劑”)檢測(cè)乳腺癌患者HER2基因狀態(tài)的臨床應(yīng)用價(jià)值。方法收集108例乳腺浸潤(rùn)性導(dǎo)管癌腫瘤組織標(biāo)本,分別采用GLP和Vysis PathVysion HER2探針試劑運(yùn)用FISH技術(shù)檢測(cè)乳腺癌患者HER2基因擴(kuò)增狀態(tài)。以IHC染色結(jié)果為參考,比較2種探針試劑檢測(cè)乳腺癌患者HER2基因狀態(tài)的差異,并評(píng)價(jià)GLP HER2探針試劑檢測(cè)乳腺癌患者組織標(biāo)本中HER2基因擴(kuò)增的敏感度、特異度、準(zhǔn)確度、陽(yáng)性預(yù)測(cè)值和陰性預(yù)測(cè)值。比較2種試劑盒不同時(shí)間段檢測(cè)同一標(biāo)本的重復(fù)性和穩(wěn)定性。結(jié)果GLP HER2探針和PathVysion HER2探針檢測(cè)乳腺癌患者組織標(biāo)本中HER2基因擴(kuò)增率分別為25.0%(27/108)和26.9%(29/108)。與PathVysion HER2探針相比,GLP HER2探針檢測(cè)乳腺癌患者組織標(biāo)本中HER2基因擴(kuò)增的敏感度、特異度和準(zhǔn)確度分別為89.7%(26/29)、98.7%(78/79)和96.3%(104/108),陽(yáng)性預(yù)測(cè)值(PPV)和陰性預(yù)測(cè)值(NPV)均為96.3%(26/27,78/81)。GLP HER2探針檢出17號(hào)染色體多倍體的敏感度、特異度和準(zhǔn)確度分別為93.3%(14/15)、100%(93/93)和99.1%(107/108)。在1個(gè)月內(nèi),均間隔5天時(shí)間,4次分別檢測(cè)了同一標(biāo)本的HER2基因狀態(tài),Vysis PathVysion HER2試劑盒檢測(cè)結(jié)果表明,4次檢測(cè)一致率為100%。GLP試劑盒的檢測(cè)結(jié)果同樣表明,4次檢測(cè)的一致率為100%,提示2種試劑盒的穩(wěn)定性和可重復(fù)性較高。結(jié)論GLP HER2探針試劑盒在檢測(cè)乳腺癌患者組織標(biāo)本中HER2基因擴(kuò)增的敏感度和特異度高,在臨床評(píng)價(jià)乳腺癌HER2基因狀態(tài)中具有廣泛應(yīng)用價(jià)值。本研究運(yùn)用免疫組織化學(xué)(Immunohistochemistry, IHC)和原位雜交技術(shù)(Fuorescence in situ hybridization, FISH)分別評(píng)價(jià)了中國(guó)不同地域和不同種族3,149例乳腺癌患者HER2的表達(dá)水平,分析了HER2表達(dá)的流行病學(xué)特征、與地理位置以及及臨床病理特征的關(guān)系,闡述了中國(guó)乳腺癌患者的病理特征與西方國(guó)家的差異。結(jié)果表明,中國(guó)乳腺癌患者發(fā)病年齡較西方早,有2個(gè)發(fā)病高峰,臨床病理特征與美國(guó)、歐洲相比,腫瘤負(fù)荷較大(p<0.001)、組織分級(jí)較差(p<0.001)、且ER/PR多為陰性(p<0.001),提示,中國(guó)乳腺癌患者的腫瘤侵襲性可能更強(qiáng)。HER2蛋白過表達(dá)率和基因擴(kuò)增率分別為23.3%和27.5%,兩者一致率達(dá)71.2%(Kappa=0.494,p<0.001)。其中,IHC檢測(cè)結(jié)果為2+的患者中,FISH檢測(cè)到72.9%為HER2基因無擴(kuò)增。隨機(jī)比對(duì)5個(gè)參與檢測(cè)的實(shí)驗(yàn)室間的HER2結(jié)果顯示,IHC和FISH檢測(cè)結(jié)果不一致率分別為5%-28%、1%-16%。HER2基因擴(kuò)增與腫瘤進(jìn)展期(p=0.002)、腫瘤直徑>5cm(p=0.002)、中度或差的組織分級(jí)(p<0.001)、絕經(jīng)(P<0.001)、ER/PR陰性(p=0.002)以及淋巴結(jié)浸潤(rùn)>4枚(p<0.001)密切相關(guān)。滿族和回族人群HER2擴(kuò)增率顯著高于其他少數(shù)民族(p<0.001)。南方與北方乳腺癌患者的HER2擴(kuò)增狀態(tài)無顯著差異(p>0.05)。本研究揭示了中國(guó)乳腺癌HER2蛋白過表達(dá)和基因擴(kuò)增流行病學(xué)特征,闡述了中國(guó)乳腺癌患者臨床病理特征與西方人群的差異,為乳腺癌個(gè)體化治療和早期預(yù)防提供有價(jià)值的理論依據(jù)。目的：本研究探討了17號(hào)染色體多倍體在判讀浸潤(rùn)性乳腺癌HER2檢測(cè)結(jié)果中的影響。目前,美國(guó)ASCO/CAP指南將FISH檢測(cè)HER2陽(yáng)性定義為單個(gè)核HER2基因拷貝數(shù)>6或HER2/CEP17比值>2.2。但這一準(zhǔn)側(cè)存在一定的矛盾,提示17號(hào)染色體多倍體可能會(huì)影響HER2基因狀態(tài)的結(jié)果判讀。方法：收集了自2010年5月至2011年8月中國(guó)醫(yī)學(xué)科學(xué)院腫瘤醫(yī)院就診的乳腺癌患者腫瘤石蠟組織標(biāo)本348例。運(yùn)用FISH技術(shù)檢測(cè)腫瘤組織HER2和CEP17的拷貝數(shù)變化。IHC檢測(cè)HER2蛋白表達(dá)狀態(tài)。結(jié)果：所有患者中CEP17半均拷貝數(shù)為2.1(1.0-12.4),17號(hào)染色體多倍體的比率(CEP17≥3)為13.8%(48/384)。HER2基因拷貝數(shù)>6的患者占26.4%(92/348),HER2/CEP17>2.2的占25.3%(88/348)。HER2拷貝數(shù)>6的患者中,17號(hào)染色體多倍體的比率占38.0%(35/92),HER2蛋白表達(dá)與HER2基因拷貝數(shù)以及比值密切相關(guān)。17號(hào)染色體多倍體與HER2蛋白表達(dá)或基因擴(kuò)增相關(guān),但相關(guān)性較差(分別為r=0.271,p<0.05和r=0.223,p<0.05)。在HER2拷貝數(shù)≤6的患者中,17號(hào)染色多倍體的比率占5.4%(5/92)。與HER2陽(yáng)性組的臨床病理特征相比,單純17號(hào)染色多倍體組的臨床病理特征更傾向于HER2陰性組的病理特征。結(jié)論：17號(hào)染色體CEP17拷貝數(shù)的增加影響了部分HER2結(jié)果的判讀。17號(hào)染色體多倍體與HER2蛋白/基因的表達(dá)相關(guān),但相關(guān)性差,且17號(hào)染色多倍體組的臨床病理特征更傾向于HER2.陰性組的病理特征,提示單純的17號(hào)染色體多倍體并不能作為HER2基因擴(kuò)增的獨(dú)立預(yù)測(cè)因子。目的：目前對(duì)血清中HER2胞外區(qū)的研究多集中在復(fù)發(fā)轉(zhuǎn)移晚期乳腺癌患者,對(duì)早期乳腺癌患者血清中HER2胞外區(qū)表達(dá)的研究甚少。本研究分析了乳腺癌血清HER2ECD表達(dá)與臨床病理特征的關(guān)系及臨床應(yīng)用價(jià)值。方法：收集中國(guó)醫(yī)學(xué)科學(xué)院腫瘤醫(yī)院Ⅰ-ⅢA期乳腺癌患者術(shù)前血以及配對(duì)腫瘤組織標(biāo)本232例,運(yùn)用酶聯(lián)免疫吸附法(ELISA)評(píng)價(jià)血清HER2ECD水平,IHC法和FISH法檢測(cè)腫瘤組織HER2表達(dá),分析乳腺癌血清HER2ECD表達(dá)水平與臨床病理特征的關(guān)系。結(jié)果：血清HER2ECD(?)的中位濃度為6.8ng/ml(范圍1.3-42.1ng/ml), HER2ECD水平升高的界值為7.4ng/ml,低于轉(zhuǎn)移性乳腺癌HER2ECD水平的臨界值15ng/ml。89例(38.3%)患者血清HER2ECD表達(dá)水平升高,而腫瘤組織77例(33.2%)HER2陽(yáng)性表達(dá),一致率達(dá)76.7%。血清HER2ECD升高與絕經(jīng)(p<0.001)、高級(jí)別組織分級(jí)(p<0.001)、ER-(p=0.007)、PR-(p<0.001)、CA153水平升高(p=0.039)和TPS水平升高(p=0.018)密切相關(guān)。結(jié)論：與單獨(dú)通過腫瘤組織檢測(cè)HER2表達(dá)狀態(tài)相比,聯(lián)合術(shù)前血清中HER2ECD水平能提供更多信息。我們支持降低術(shù)前血清中HER2ECD (?)臨界值指標(biāo)更有利于評(píng)價(jià)早期乳腺癌患者腫瘤組織HER2的狀態(tài)。

    Molecular techniques play an increasingly important role in breast cancer detection and help in the prediction of prognosis and treatment response. According to the receptors status to choose the endocrine therapy and molecular targeted therapy for breast cancer patients is the model of the individual treatment. Especially, epidermal growth factor receptor2(HER2) is an important therapeutic target for breast cancer treatment. Trastuzumab (Herceptin(?), Roche) is a recombinant humanised monoclonal antibody directed against the extracellular domain of the HER2protein that significantly increases the survival of HER2positive patients, both as monotherapy or in combination with chemotherapy. Furthermore, HER2positive tumors are more sensi tive to anthracyclin chemotherapy and seem to respond better to aromatase inhibitors than to tamoxifen. Given its prognostic, predictive and therapeutic impli-cations, an accurate assessment of HER2status for breast cancer patients is crucial.HER2, a185KDa transmemb rane receptor, is a proto-oncogene located on the long arm of chromosome17that encodes a transmembrane receptor protein of the epidermal growth factor receptor family. The tyrosine kinase domains are activated by both homodimerization and heterodimerization. Transcription factors activated by the pathway regulate many genes involved in cell proliferation, survival, differentiation, angiogenesis, and invasion and metastasis. However, with the development of HER2testing and clinical application of trastuzumab in China, a number of problems have cropped up in this field as followed.1. There are many techniques to evaluate HER2status, but the standardization of techniques, the accuracy of detection results, the concordance of different methods, and the expensive reagent from aboard are also controversial.2. The epidermiology of HER2expression of breast cancer patients in China is not clear.3. The incidence of chromosome17polysomy and the association with HER2overexpression is also not clear.4. Whether the serum is a surrogate for tissues to response HER2expression.According to the questions above, this study focused on four parts as followed,Part1. Study on comparison of GLP and PathVysion FISH assays on measuring HER2gene status in breast cancer. This study detected HER2gene status from108cases with invasive ductal breast cancer using GLP and PathVysion HER2probe kits by FISH, compared HER2gene expression differentiation measured by GLP and PathVysion HER2probe, and assessed the sensitivity, the specificity and the accuracy of GLP HER2probe kit. HER2gene amplification cases were25.0%(27/108) and26.9%(29/108) detected by GLP probe kit and PathVysion probe kit, respectively. As assessed with PathVysion HER2probe as the reference, the sensitivity, the specificity and the accuracy of the GLP HER2kit were calculated to be89.7%(26/29),98.7%(78/79) and96.3%(104/108), respectively, whereas the positive predictive value (PPV) and negative predictive value (NPV). GLP probe kit can be considered to be high sensitivity and specificity, and it has the widespread clinical application value in assessing HER2gene status for breast cancer patients.Part2. The epidermiological characteristics of HER2expression in Chinese breast cancer. We performed a multicenter study on the HER2status in3149breast cancer specimens from different ethnic populations and areas in China by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) assays. The positive rates for HER2over-expression and HER2amplification were23.3%and27.5%in this population, respectively. The concordance between IHC and FISH was71.2%(Kappa=0.494, p<0.001). The HER2amplification was associated significantly with advanced tumor stage (Ⅲ or Ⅳ)(p=0.002), large tumor size (>5cm)(p=0.002), moderately and poorly histological grades (p<0.001), postmenopause (p<0.001), ER/PR-(p=0.002), and having≥4lymph nodes affected (p<0.001) in this population. The positive rates of HER2amplification in specimens from Man and Hui Chinese were significantly higher than that in other Chinese populations. There are slightly higher positive rates of HER2expression and amplification in Chinese patients with breast cancer. These findings may provide new insights into understanding the epidemiological features of HER2expression and amplification, and may be valuable for clinical practice.Part3. Impact of polysomy17on HER2testing of breast cancer. We collected384cases of breast cancer collected from Cancer Institute/Hospital of Chinese Academy of Medical Sciences&Peking Union Medical College. Chromosome17copies and HER2gene status were identified by fluorescent in situ hybridization, and the corresponding HER2immunohistochemistry was obtained. The average CEP17copy number for the group was2.1(range,1.0-12.4). Fourty-eight cases (13.8%,48/348) were idenfied as chromosome17polosomy with CEP17copy number≥3. Ninety-two (26.4%) cases had>6copies of HER2per nucleus, and95cases (25.3%) qualified as HER2gene amplified using the HER2/CEP17ratio (>2.2) guideline. All these cases had>6HER2signals, presented38.0%cases with>3CEP17copy number. HER2protein expression showed significant positive correlations with both HER2gene copy number and HER2/CEP17ratio (p<0.01, r=0.56and0.64, respectively). In conclusion, increased CEP17signals detected in invasive breast carcinomas may lead to discordant interpretation of HER2gene amplification in a significant proportion of the cases, depending on which criterion (ratio vs absolute number) is used for interpretation. However, increased gene dosage (>6HER2genes or HER2/CEP17ratio>2.2), regardless of the evaluation method, is positively correlated with HER2protein expression. The polysomy of chromosome17can not be a independent predictive factor for HER2gene amplification.Part4. Relationship of serum HER2level and tissue HER2expression in early stage of breast cancer. The measurement of the HER2protein in the serum of metastatic breast cancer patients has now been reported, but there are no consistent data to support the clinical utility of serum HER2extracellular domain (ECD) for patients with early breast cancer. We aimed to evaluate the correlation between serum HER2ECD levels and tumor HER2status, and analyze their relationship with clinicopathological parameters in patients with early stage disease. This study was conducted on232breast cancer patients with stage Ⅰ-ⅢA diseases before treatment. Preoperative serum samples were measured by enzyme-linked immunosorbent assay. Tissue HER2status was analyzed by immunohistochemistry and fluorescence in situ hybridization assays. The median serum HER2ECD concentration was6.8ng/ml. The best diagnostic cut-off value was7.4ng/ml, with62.9%sensitivity and85.3%specificity. High serum HER2ECD levels were reported in89patients (38.3%) and HER2tissue positive expression was observed in77patients (33.2%) with a moderate concordance of76.7%. Elevated serum HER2ECD correlated with postmenopausal (p<0.001), high tumor grade (p<0.001), negativity of both estrogen (p=0.007) and progesterone receptors (p<0.001),high level of CA153(p=0.039) and TPS (p=0.018). HER2ECD may provide more additional information compared with HER2tissue alone. We supported that it is necessary to decrease the cut-off value in evaluating serum HER2ECD level for early stage of breast cancer.Taken together, we cleared the wide application value of GLP HER2DNA probe kit, got the epidemiological data of HER2expression in Chinese breast cancer patients, found the influence of chromosome17polysomy on HER2status, and explicated the feasibility of serum instead of tissue to detect HER2expression in new diagnosis breast cancer, which may provide the basis for accurately interpreting HER2status in the future clinical practice, and promot individualized targeted therapy in clinical application. Objective To evaluate clinical application of Jin Pujia GLP HER2probe kit in testing HER2gene status of breast cancer through comparing with Vysis PathVysion HER2probe kit. Methods This study detected HER2gene status from108cases with invasive ductal breast cancer using GLP and PathVysion HER2probe kits by FISH, compared HER2gene expression differentiation measured by GLP and PathVysion HER2probe, and assessed the sensitivity, the specificity and the accuracy of GLP HER2probe kit. Results HER2gene amplification cases were25.0%(27/108) and26.9%(29/108) detected by GLP probe kit and PathVysion probe kit, respectively. As assessed with PathVysion HER2probe as the reference, the sensitivity, the specificity and the accuracy of the GLP HER2kit were calculated to be89.7%(26/29),98.7%(78/79) and96.3%(104/108), respectively, whereas the positive predictive value (PPV) and negative predictive value (NPV) were96.3%(26/27) and96.3%(78/81), respectively. With regard to the ability to presume polysomy17, the GLP HER2kit had a sensitivity of93.3%(14/15), a specificity of100%(93/93) and an accuracy of99.1%(107/108). Each lot was run on five non-consecutive days over one month. The GLP and PathVysion HER2probe kit all had high reproducibility with concordance rate of100%. Conclusion GLP probe kit can be considered to be high sensitivity and specificity, and it has the widespread clinical application value in assessing HER2gene status for breast cancer patients. We performed a multicenter study on the HER2status in3149breast cancer specimens from different ethnic populations and areas in China by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) assays. The potential association of HER2status with demographic and clinical characteristics was analyzed. The positive rates for HER2over-expression and HER2amplification were23.3%and27.5%in this population, respectively. The concordance between IHC and FISH was71.2%(Kappa=0.494, p<0.001). Furthermore,72.9%of specimens with IHC2+were negative to FISH. The discordance rates among laboratories were from5%to28%for IHC and1%to16%for FISH. The HER2amplification was associated significantly with advanced tumor stage (Ⅲ or Ⅳ)(p=0.002), large tumor size (>5cm)(p=0.002), moderately and poorly histological grades (p<0.001), postmenopause (p<0.001), ER-PR-(p=0.002), and having≥4lymph nodes affected (p<0.001) in this population. The positive rates of HER2amplification in specimens from Man and Hui Chinese were significantly higher than that in other Chinese populations. There are slightly higher positive rates of HER2expression and amplification in Chinese patients with breast cancer than that in Caucasian. These findings indicate the epidemiological features of HER2expression in Chinese breast cancer patients, state the difference between Chinese patients and Western Caucasian patients, which may provide new insights into understanding personalized targeted therapy for breast cancer, and may be valuable for early prevention in clinical practice. Backgroud:The current study was performed to determine the impact of polysomy17on the interpretation of HER2testing of invasive breast carcinomas using fluorescent in situ hybridization methods. Current American Societyof Clinical Oncology/College of American Pathologists guidelines define HER2positive tumors as those with>6HER2genes per nucleus or those with HER2/CEP17(chromosome17) ratio>2.2. These guidelines are potentially con-tradictory in tumors with polysomy of chromosome17. Methods:Chromosome17copies (≥3CEP17signals on average) and HER2gene status were identified by fluorescent in situ hybridization in the384cases of breast cancer collected from Cancer Institute/Hospital of Chinese Academy of Medical Sciences&Peking Union Medical College, and the corresponding HER2immunohistochemistry was obtained. Results:The average CEP17copy number for the group was2.1(range,1.0-12.4). Fourty-eight cases (13.8%,48/348) were idenfied as chromosome17polosomy with CEP17copy number≥3. Ninety-two (26.4%) cases had>6copies of HER2per nucleus, and95cases (25.3%) qualified as HER2gene amplified using the HER2/CEP17ratio (>2.2) guideline. All these cases had>6HER2signals, presented38.0%cases with>3CEP17copy number. HER2protein expression showed significant positive correlations with both HER2gene copy number and HER2/CEP17ratio (p<0.01, r=0.56and0.64, respectively). Conclusions:Increased CEP17signals detected in invasive breast carcinomas may lead to discordant interpretation of HER2gene amplification in a significant proportion of the cases, depending on which criterion (ratio vs absolute number) is used for interpretation. Increased gene dosage (>6HER2genes or HER2/CEP17ratio>2.2), regardless of the evaluation method, is positively correlated with HER2protein expression, but the polysomy of chromosome17can not be a independent predictive factor for HER2gene amplification. Background:The measurement of the human epidermal growth factor receptor2(HER2) protein in the serum of metastatic breast cancer patients has now been reported, but there are no consistent data to support the clinical utility of serum HER2extracellular domain (ECD) for patients with early breast cancer. We aimed to evaluate the correlation between serum HER2ECD levels and tumor HER2status, and analyze their relationship with clinicopathological parameters in patients with early stage disease. Methods:A prospective study was conducted on232breast cancer patients with stage Ⅰ-Ⅱ diseases before treatment. Preoperative serum samples were measured by enzyme-linked immunosorbent assay (ELISA). Tissue HER2status was analyzed by immunohistochemistry and fluorescence in situ hybridization assays. Results:The median serum HER2ECD concentration was6.8ng/ml (range1.3-42.1ng/ml). The best diagnostic cut-off value was7.4ng/ml, with72.9%sensitivity and85.3%specificity, which was lower than HER2ECD cut-off value with metastasis breast cancer. High serum HER2ECD levels were reported in89patients (38.3%) and HER2tissue positive expression was observed in77patients (33.2%) with a moderate concordance of76.7%. Elevated serum HER2ECD correlated with postmenopausal (p<0.001), high tumor grade (p<0.001), negativity of both estrogen (p=0.007) and progesterone receptors (p<0.001), high level of carbohydrate antigen153(CA153)(p=0.039) and tissue polypeptide specific antigen (TPS)(p=0.018). Conclusion: HER2ECD may provide more additional information compared with HER2tissue alone. We support that it is necessary to decrease the cut-off value in evaluating serum HER2ECD level for early stage of breast cancer.

中國(guó)乳腺癌HER2基因檢測(cè)、臨床病理特征分析及流行病學(xué)研究

表索引5-6圖索引6-7縮略詞索引7-9中文摘要9-12Abstract12-16引言16-21    參考文獻(xiàn)18-21第一部分 GLP和PathVysion FISH探針試劑盒檢測(cè)乳腺癌HER2基因狀態(tài)的比較研究21-42    前言23-24    材料與方法24-32    結(jié)果32-36    討論36-39    小結(jié)39-40    參考文獻(xiàn)40-42第二部分 中國(guó)乳腺癌HER2表達(dá)的流行病學(xué)研究及臨床病理特征分析42-71    前言44-46    材料與方法46-49    結(jié)果49-62    討論62-66    小結(jié)66-67    參考文獻(xiàn)67-71第三部分 17號(hào)染色體多倍體對(duì)乳腺癌HER2基因狀態(tài)的影響71-95    前言73-74    材料與方法74-76    結(jié)果76-85    討論85-89    小結(jié)89-90    參考文獻(xiàn)90-95第四部分 乳腺癌血清中HER2 ECD的表達(dá)研究95-121    前言97-98    材料與方法98-102    結(jié)果102-110    討論110-115    小結(jié)115-116    參考文獻(xiàn)116-121基金資助121-122文獻(xiàn)綜述122-139    參考文獻(xiàn)132-139個(gè)人簡(jiǎn)歷139-142在讀期間發(fā)表文章142-173致謝173-174

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