人乳腺癌裸鼠移植瘤组织Duffy抗原趋化因子受体与Her-2蛋白表达相关性的探讨

目的:探讨人乳腺癌裸鼠移植瘤组织Duffy抗原趋化因子受体(DARC)与Her-2蛋白表达的相关性.方法:建立人乳腺癌裸鼠原位移植瘤模型,按细胞类型分为DARC低表达的MDA-MB-231裸鼠移植瘤、转染空质粒的MDA-MB-231(MDA-MB-231-vect)裸鼠移植瘤及转染DARC cDNA的MDA-MB-231(MDA-MB-231-DARC)裸鼠移植瘤.隔天观察裸小鼠的健康状况和肿瘤细胞的成瘤性,成瘤后每周测量1次各移植瘤的体积,至第2周及第4周时,分别处死各组实验动物8只,获取移植瘤,免疫组化检测原位移植瘤组织中DARC及Her-2蛋白表达;选取中国福利会国际和平妇幼保健院乳腺癌临床组织标本80例,通过免疫组化技术检测DARC和Her-2蛋白表达,并对其相关性进行分析.结果:虽然各组动物的成瘤率均为100%,但DARC转染后的 MDA-MB-231移植瘤(MDA-MB-231-DARC移植瘤)生长明显减慢(P=0.007),且裸鼠移植瘤组织中Her-2蛋白呈低表达,转染与未转染DARC的裸鼠移植瘤组织中Her-2蛋白表达差异有统计学意义,χ2=69.83,P=0.045;对人乳腺癌临床组织标本免疫组化检测结果显示,Her-2蛋白表达与DARC蛋白呈负相关,r=-0.464,P=0.026.结论:人乳腺癌裸鼠移植瘤组织中DARC的过表达抑制Her-2蛋白表达.     

表达白细胞介素-12质粒DNA抗小鼠肝癌皮下移植瘤的作用

目的 探讨瘤内注射mIL-12质粒DNA抗小鼠肝癌皮下移植瘤的作用。方法：构建真核表达质粒载体pDC511mIL-12，ELISA方法检测质粒载体在真核细胞中的表达，淋巴母细胞增殖法检测mIL-12的生物学活性；分别于小鼠肝癌H22皮下移植瘤内直接注射质粒DNA，观察各组小鼠存活时间、肿瘤体积变化及各组小鼠脾脏细胞毒T淋巴细胞(CTL)的活性：注射质粒DNA后1月进行瘤体组织病理学观察：结果：mIL-12基因治疗组与空载体对照组相比，肿瘤生长显著受抑制(F=4．10，P=0．03)，小鼠存活期显著延长(X^2=4．48，P=0．03)．并且小鼠脾细胞CTL杀伤活性增强。质粒DNA瘤内注射1月后，pDC511mIL-12组肿瘤病灶炎性细胞浸润明显，病灶内肿瘤细胞广泛坏死。结论：瘤内注射mIL-12表达质粒DNA可抑制小鼠肝癌皮下移植瘤生长，能提高机体抗肿瘤免疫应答。     

E1B缺陷腺病毒对鼻咽癌CNE-2细胞杀伤作用及其机理研究

目的：研究E1B缺陷腺病毒d11520对鼻咽癌CNE-2细胞的杀伤作用及其作用机理。方法：用MTT法和细胞病变试验（CPE）测量d11520在体外对CNE-2细胞的抑制和杀伤作用。并通过测定病毒在CNE-2细胞中的复制产量和DAPI染色试验探讨其杀伤机理；瘤内注射d11520，观察其对CNE-2裸鼠移植瘤的治疗效果。用免疫织化方法检测肿瘤组织中病毒的复制。结果：体外实验结果显示：d11520能在CNE-2细胞中复制，导致明显的细胞病变，显著抑制CNE-2细胞的生长，在病变细胞中可见明显的核膨胀，瘤内注射d11520能明显抑制裸鼠移植瘤的生长，在肿瘤组织中可检测到病毒复制。结论：d11520能在CNE-2细胞中复制并使之裂解，从而在体内外有效地抑制和杀伤CNE-2细胞。     

过表达CNTN1对裸鼠乳腺癌转移瘤生长的影响

目的:探讨过表达CNTN1对乳腺癌Hs578T细胞裸鼠皮下移植瘤生长的促进作用,为乳腺癌生物治疗提供实验依据.方法:脂质体法介导pEGFP-N1-CNTN1真核表达载体转染入乳腺癌Hs578T细胞,G418筛选出稳定表达CNTN1的Hs578T细胞(Hs578T-CNTN1细胞);接种Hs578T-CNTN1细胞制备裸鼠皮下移植瘤模型,观察CNTN1过表达对Hs578T细胞移植瘤生长的影响.结果:Western blot结果显,转染pEGFP-N1-CNTN1组Hs578T细胞中CNTN1蛋白表达量高于pEGFP-N1组及未转染组.Hs578T-CNTN1、Hs578T-N1和Hs578T细胞接种裸鼠的第20天,Hs578T-CNTN1组移植瘤质量较Hs578T组和Hs578T-N1组移植瘤质量显著增加[(4.62 ±0.22)g,(2.56 ±0.76)g和(2.10±0.78)g,分别P＜0.01和P＜0.05].结论:CNTN1过表达可以促进乳腺癌Hs578T细胞裸鼠移植瘤的生长.     

慢病毒介导双自杀基因对乳腺癌细胞的体内杀伤作用

目的:研究慢病毒介导的KDR启动子驱动的胞嘧啶脱氨酶(CD)/胸苷激酶(TK)融合基因系统(FGW-KDRP-CD/TK)对乳腺癌细胞的体内杀伤作用.方法:培养乳腺癌MCF-7细胞,建立裸鼠荷瘤模型.荷瘤后将裸鼠随机分为4组.Ⅰ组:空白对照,荷瘤但不施加任何处理;Ⅱ组:注射慢病毒与前药(5-FC+GCV);Ⅲ组:仅注射慢病毒;Ⅳ组:仅注射前药.观察肿瘤生长速度,测量瘤体大小及重量;用RT-PCR法鉴定双自杀基因在转基因瘤组织中的表达;取肿瘤组织进行HE及PCNA免疫组化染色;流式细胞技术进行细胞周期检测.结果:第Ⅱ组裸鼠移植瘤的生长显著受到抑制,第Ⅰ、Ⅲ、Ⅳ组肿瘤生长情况无明显差异.经RT-PCR检测发现转基因瘤组织有目的基因的表达.与第Ⅰ组(空白对照组)相比,第Ⅱ组瘤组织的PCNA表达明显下调.流式细胞仪检测细胞周期分析显示治疗后G1期细胞比率增多,G2/M期细胞减少.结论:FGW-KDRP-CD/TK联合前药5-FC及GCV在体内可明显抑制移植瘤的生长,细胞周期阻滞,该作用可能与抑制PCNA表达有关.     

黏蛋白1基因转染DC对乳腺癌细胞MCF-7裸鼠移植瘤的免疫抑制作用

目的:探讨黏蛋白1 (mucin 1,MUC1基因转染DC对人乳腺癌MCF-7细胞裸鼠移植瘤的抑制作用.方法:体外诱导培养健康成人DC,应用脂质体转染法将pcDNA3.1-MUC1转染DC,ELISA法检测转染后DC分泌细胞因子IL-12和YNF-α的能力,LDH释放法检测基因转染后DC诱导特异性CTL对乳腺癌MCF-7细胞的杀伤活性.应用MU C1基因转染DC、空质粒转染DC、及生理盐水皮下注射治疗人乳腺癌MCF-7细胞裸鼠移植瘤,观测其对肿瘤生长的抑制作用.结果:转染pcDNA3.1-MUC1的DC分泌IL-12、TNF-α的能力较转染空质粒DC明显增强[IL-12:(202.52±29.61)vs(10.83±1.02)pg/ml;TNF-α:(349.07±79.42)vs(9.26±1.52)pg/ml,均P＜0.01];转染pcDNA3.1-MUC1的DC诱导产生特异性CTL,对人乳腺癌MCF-7细胞具有更明显的杀伤活性,效靶比为10∶1、5∶1和2.5∶1时的杀伤率分别达到56.2％、38.9％和25.8％,显著高于对照组CTL(均P＜0.01).MUC1基因转染DC对乳腺癌MCF-7裸鼠移植瘤生长抑制作用明显强于空质粒转染DC组(P＜0.05).结论:MUC1基因转染DC可以诱导特异性CTL,对乳腺癌MCF-7细胞具有更强的抗肿瘤免疫效应.     

转染REGγ基因对乳腺癌细胞裸鼠移植瘤生长的影响

目的:研究转染蛋白酶体激活因子REGγ基因的乳腺癌MDA-MB-231细胞在裸鼠体内的成瘤作用及其机制.方法:以脂质体转染法将构建的重组质粒pcDNA3.1-REGγ导入MDA-MB-231细胞,以G418(600 mg/L)筛选获得稳定高表达该基因的细胞株(实验组).以转染空载体及未施加处理因素的细胞作为对照组.将此3组细胞接种于裸鼠皮下,观察移植瘤的生长情况并计算抑瘤率.RT-PCR检测REGγ基因在移植瘤中的表达,FCM检测移植瘤的肿瘤浸润淋巴细胞(tumor-infiltrating lymphocyte,TIL)中CD16的表达以及肿瘤细胞周期和细胞凋亡,免疫组织化学法检测移植瘤中p21的表达.结果:与对照组相比,实验组的移植瘤生长速度较快、体积较大,瘤体质量明显增加(P＜0.05);REGγ基因在移植瘤中的表达增加(P＜0.01);FCM检测提示CD16阳性率明显降低(P＜0.05);G0/G1和G2/M期细胞减少,S期细胞明显增多,肿瘤细胞的凋亡率明显降低(P＜0.05);p21的表达明显降低(P＜0.05).结论:REGγ基因在体内具有促进乳腺癌发生、发展的作用,其机制可能与加速细胞周期、抑制细胞凋亡、抑制自然杀伤细胞活化以及对p21的特异性降解有关.     

DC 荷载α－GalCer 联合肿瘤特异性 CTL 对小鼠 Heps 肝癌移植瘤生长的影响

目的：探讨树突状细胞（DC）荷载α－半乳糖神经酰胺（α－GalCer）联合肿瘤特异性细胞毒 T 淋巴细胞（CTL）对小鼠 Heps 肝癌移植瘤生长的抑制作用。方法：诱导扩增小鼠骨髓来源的 DC 细胞和 T 淋巴细胞,培养成为具有肿瘤特异性的 CTL,DC 细胞体外荷载α－GalCer。建立 Heps 肝癌移植瘤模型,将36只模型鼠随机分为4组（n ＝9）,分别尾静脉给予生理盐水（对照组）、CTL（CTL 组）、DC 荷载α－GalCer（DC 荷载α－GalCer 组）、DC 细胞荷载α－GalCer ＋CTL（联合治疗组）输注。每隔两天测量移植瘤体积,观察体积变化。于治疗后第14d,处死小鼠,取眼球血及瘤体组织。流式细胞术（FCM）检测外周血 CD4＋、CD8＋ T 细胞比例。免疫组织化学染色观察肿瘤组织中 Bcl －2／Bax 凋亡蛋白的表达。结果：与对照组相比,各治疗组均可抑制肿瘤的生长,差异具有统计学意义（P ＜0．01）;联合治疗组较 DC 荷载α－GalCer 组及 CTL 组肿瘤体积明显减小（P ＜0．05）。联合治疗组小鼠外周血 CD4＋、CD8＋ T 细胞数量较另外三组显著升高（P ＜0．05）;对照组、CTL组、DC 荷载α－GalCer 组小鼠外周血中 CD4＋、CD8＋ T 细胞含量无明显差异（P ＞0．05）。与对照组相比,各治疗组移植瘤内 Bax 阳性细胞数量明显增加（P ＜0．05）,Bcl －2阳性细胞数量明显减少（P ＜0．05）;与 CTL组和α－GalCer 组相比,联合治疗组移植瘤内 Bax 阳性细胞明显增加（P ＜0．05）,Bcl －2阳性细胞明显下降（P ＜0．05）。结论：DC 荷载α－GalCer 与肿瘤特异性 CTL 联合应用能够对小鼠 Heps 肝癌移植瘤具有协同杀伤作用。     

选择性增殖腺病毒CNHK500治疗乳腺癌的实验研究

目的: 观察选择性增殖腺病毒CNHK500对乳腺癌的特异性杀伤作用.方法: 行病毒增殖实验和细胞生长抑制实验,验证CNHK500选择性复制和杀伤能力; Western blot检测腺病毒E1A和E1B在细胞中的表达.结果: CNHK500在乳腺癌细胞中复制能力与野生型腺病毒wtAd5相似,较ONYX-015增殖能力强.在正常成纤维细胞中CNHK500病毒增殖能力明显减弱, 较wtAd5增殖能力弱1 000倍左右.CNHK500可有效杀伤乳腺癌细胞株;而CNHK500对正常成纤维细胞的杀伤力较wtAd5减弱约100倍.CNHK500病毒的E1A可以选择性在端粒酶阳性的乳腺癌细胞株中表达,在端粒酶阴性的正常成纤维细胞株BJ中不表达, CNHK500可以选择性地在缺氧条件下表达E1B.动物实验结果显示,静脉注射CNHK500可以显著抑制MCF-7乳腺癌细胞裸鼠移植瘤的生长,治疗效果与给药剂量相关.结论: 肿瘤选择性增殖腺病毒CNHK500可选择性在端粒酶阳性的乳腺癌细胞中复制,并产生体内外杀伤作用.     

EGFRvⅢ/CAR-T对EGFRvⅢ + U87胶质瘤细胞和裸鼠移植瘤的特异性 杀伤作用

目的： 制备靶向EGFRvⅢ的第三代CAR-T(EGFRvⅢ/3CAR-T),检测其在体外和体内对EGFRvⅢ + U87胶质瘤细胞的 特异性杀伤效应。 方法： 用磷酸钙沉淀法将三质粒共转染HEK293T细胞包装EGFRvⅢ/3CAR慢病毒（LV-EGFRvⅢ/3CAR）,感 染健康人CD3 + T细胞,Western blotting和流式细胞术检测T细胞中EGFRvⅢ/3CAR的表达水平, 51 Cr释放法检测EGFRvⅢ/3CAR- T对EGFRvⅢ + U87细胞的体外杀伤效应,ELISA法检测EGFRvⅢ/3CAR + T的IFN-γ分泌水平。构建裸鼠异种胶质瘤移植模型, 检测EGFRvⅢ/3CAR-T对移植瘤的体内杀伤活性。 结果： EGFRvⅢ/3CAR慢病毒包装成功,滴度值为5×10 6 TU/ml。Western blotting在相对分子质量58 000处检测到EGFRvⅢ/3CAR表达,未转导组无相同分子量蛋白表达。流式细胞术检测结果显示EG- FRvⅢ/3CAR转导效率平均为52.3%, 51 Cr释放法检测EGFRvⅢ/3CAR-T特异性杀伤作用与E∶T值（E∶T为4∶1、为8∶1、16∶1、32∶ 1）呈正比关系。ELISA法检测到细胞因子IFN-γ分泌量为(1 836±148.2) pg/ml,与NT T和GFP + T细胞相比差异有统计学意义 （ P ＜0.01）。EGFRvⅢ/3CAR + T细胞的特异性杀伤活性及IFN-γ分泌均依赖于EGFRvⅢ在U87细胞中的表达水平。体内肿瘤生 长检测结果显示,注射后3周EGFRvⅢ/3CAR + T组肿瘤体积较GFP + T细胞和PBS组差异有统计学意义（ P< 0.01）。 结论： EGFRv Ⅲ/3CAR-T在体外和体内均能发挥靶向杀伤EGFRvⅢ + 脑胶质瘤细胞的作用,为后续临床试验提供依据。     

Expression of Her-2/neu in human lung cancer cell lines by immunohistochemistry and fluorescence in situ hybridization and its relationship to in vitro cytotoxicity by trastuzumab and chemotherapeutic agents.

Overexpression of the Her-2/neu oncogene and receptor protein was reported in approximately 20% of breast cancers and was associated with a poor prognosis. Her-2/neu expression was a predictor for response to trastuzumab, a monoclonal antibody that recognizes the Her-2/neu cell surface receptor. Data regarding the expression of Her-2/neu in lung cancer are far more limited, and there is little information regarding the influence of Her-2/neu expression and response to trastuzumab alone or in combination with chemotherapeutic agents. In this report we evaluated Her-2/neu gene expression by fluorescence in situ hybridization (FISH) and the cell surface expression of the Her-2/neu receptor by immunohistochemistry using the HercepTest and by FACS analysis in 31 lung cancer cell lines with 5 breast cancer cell lines as controls. By FACS, we found Her-2/neu overexpression (mean fluorescence intensity >8) in 2 of the 22 non-small cell lung cancer (NSCLC) cell lines (9%), none of 11 small cell lung cancer (SCLC) cell lines, and 4 of 5 breast cancer cell lines. A positive HercepTest (2+ or 3+) was found in 6 of 19 NSCLC cell lines (26%, 2+; 5%, 3+), 1 of 3 SCLC cell lines (33%), and 4 of 5 breast cancer cell lines (80%). One of 6 NSCLC cell lines examined (17%) had gene amplification with >32 copies of Her-2/neu/cell and had homogeneous staining regions. One NSCLC cell line had a maximum of 14 copies of Her-2/neu/cell, and 3 had modest increases in Her-2/neu gene copy number without gene amplification (maximum 5-8 copies/cell). None of the SCLC cell lines had more than a maximum of 4 copies/cell, whereas the 2 breast cancer cell lines had maximum Her-2/neu copy numbers of 80 and 5, respectively. Aneusomy rather than true amplification was the major cause of increased Her-2/neu expression in most of the NSCLC cell lines. There was a strong correlation when the results of fluorescence-activated cell sorter, HercepTest results, and FISH were compared in pairs. Furthermore, Trastuzumab produced a G(1) cell cycle arrest and growth inhibition only in cell lines expressing Her-2/neu. The IC(50) for growth inhibition was correlated with cell surface Her-2/neu expression. The combination of trastuzumab and chemotherapeutic agents produced more than additive growth inhibition in cell lines expressing Her-2/neu, but the level of additivity was not related to the amount of Her-2/neu expression. These data indicate that trastuzumab alone and in combination with chemotherapeutic agents should be tested in NSCLC patients and that Her-2/neu should be assessed by both immunohistochemistry and FISH methods in these studies to determine which test is the best predictor of outcome.     

激素受体和人表皮生长因子受体2的表达与改良根治术后淋巴结阳性乳腺癌患者预后的关系

目的 探讨雌激素受体(ER)、孕激素受体(PR)和人表皮生长因子受体2(Her-2)的表达情况与行改良根治术后腋窝淋巴结阳性乳腺癌患者预后的关系.方法 收集835例行改良根治术后腋窝淋巴结阳性乳腺癌患者的临床和随访资料.根据ER、PR和Her-2的免疫组化检查结果,将患者分为Rec-/Her-2-(三阴性)组、Rec-/Her-2+组、Rec+/Her-2+组和Rec+/Her-2-组,比较其局部区域复发率、远处转移率、无瘤生存率和总生存率.结果 835例患者中,三阴性组141例,Rec-/Her-2+组99例,Rec+/Her-2+组157例,Rec+/Her-2-组438例.Rec+/Her-2-患者的5年局部区域复发率为6.2%,低于其他患者(12.9%,P=0.004).与受体阳性组(Rec+/Her-2+和Rec+/Her-2-)比较,受体阴性组(Rec-/Her-2-和Rec-/Her-2+)有较高的5年远处转移率(26.4%和19.7%,P=0.0008)、较低的5年无瘤生存率(66.7%和75.6%,P=0.0001)和较低的5年总生存率(71.4%和84.2%.P=0.0000).多因素Cox回归分析结果显示,激素受体和Her-2的表达状态是乳腺癌患者局部区域复发、远处转移、无瘤生存和总生存的独立影响因素(均P<0.05),Rec+/Her-2-患者的局部区域复发风险低,受体阴性患者发生远处转移和死亡的风险高.结论 ER、PR和Her-2是改良根治术后腋窝淋巴结阳性乳腺癌患者的独立预后因素.     

乳腺癌合并2型糖尿病患者沉默信息调节因子1表达及其意义

 【摘要】　目的　研究沉默信息调节因子1（SIRT1）在乳腺癌合并2型糖尿病中的表达,分析其与乳腺癌合并2型糖尿病患者相关临床病理指标之间的关系。方法　应用免疫组织化学方法检测30例乳腺癌合并2型糖尿病患者、65例非糖尿病乳腺癌患者的乳腺癌组织和18例正常乳腺组织中SIRT1的表达情况。结果　SIRT1在非糖尿病、合并2型糖尿病乳腺癌组织和正常乳腺组织中的表达率分别为76.9 %（50／65）、50.0 %（15／30）、5.6 %（1／18）,乳腺癌组织均高于正常乳腺组织（χ2＝24.618,P＝0.000）,合并2型糖尿病者SIRT1的表达率低于非糖尿病患者（χ2＝6.886,P＝0.009）。合并2型糖尿病的乳腺癌患者SIRT1的表达与淋巴结转移（P＝0.011）、pTNM分期（P＝0.028）、p53蛋白的表达（P＝0.003）以及Her-2的表达（P＝0.031）均呈正相关。结论　SIRT1在乳腺癌合并2型糖尿病患者乳腺癌组织中过表达,但阳性表达率低于未合并糖尿病乳腺癌组,其可能是影响糖尿病病程进展的重要分子; SIRT1的表达与多项临床病理指标有关,可能成为判断合并2型糖尿病乳腺癌恶性程度及评估预后的生物学指标。     

Pharmacological inhibition of fatty acid synthase (FAS): a novel therapeutic approach for breast cancer chemoprevention through its ability to suppress Her-2/neu (erbB-2) oncogene-induced malignant transformation

We designed our experiments to evaluate whether fatty acid synthase (FAS), a lipogenic enzyme linked to tumor virulence in population studies of human cancer, is necessary for the malignant transformation induced by Her-2/neu (erbB-2) oncogene, which is overexpressed not only in invasive breast cancer but also in premalignant atypical duct proliferations and in ductal carcinoma in situ of the breast. To avoid the genetic complexities associated with established breast cancer cell lines, we employed NIH-3T3 mouse fibroblasts engineered to overexpress human Her-2/neu coding sequence. NIH-3T3/Her-2 cells demonstrated a significant upregulation of FAS protein expression, which was dependent on the upstream activation of mitogen-activated protein kinase and phosphatidylinositol 3'-kinase/AKT pathways. Remarkably, pharmacological FAS blockade using the mycotoxin cerulenin or the novel small compound C75 completely suppressed the state of Her-2/neu-induced malignant transformation by inhibiting the ability of NIH-3T3/Her-2 cells to grow under either anchorage-independent (i.e., to form colonies in soft agar) or low-serum monolayer conditions. Moreover, NIH-3T3/Her-2 fibroblasts were up to three times more sensitive to chemical FAS inhibitors relative to untransformed controls as determined by MTT-based cell viability assays. In addition, pharmacological FAS blockade preferentially induced apoptotic cell death of NIH-3T3/Her-2 fibroblasts, as determined by an ELISA for histone-associated DNA fragments and by the terminal deoxynucleotidyltransferase (TdT)-mediated nick end labeling assay (TUNEL). Interestingly, the degree of Her-2/neu oncogene expression in a panel of breast cancer cell lines was predictive of sensitivity to chemical FAS inhibitors-induced cytotoxicity, while low-FAS expressing and chemical FAS inhibitors-resistant MDA-MB-231 breast cancer cells became hypersensitive to FAS blockade when they were engineered to overexpress Her-2/neu. Our observations strongly suggest that inhibition of FAS activity may provide a new molecular avenue for chemotherapeutic prevention and/or treatment of Her-2/neu-related breast carcinomas.     

乳腺癌术后放疗患者T淋巴细胞亚群与ER、PR、Her-2表达的相关性研究

目的研究T淋巴细胞亚群在乳腺癌术后患者放疗前、后的变化与ER、PR、Her-2表达之间的相关性,为临床判断预后以及乳腺癌术后治疗提供一定的参考依据。方法采用流式细胞仪BD FACS Calibur检测46例乳腺癌术后患者静脉血中淋巴细胞亚群,包括T细胞、Th细胞、Ts细胞、NK细胞以及CD4+/CD8+比值,并用四色荧光进行分析。采用免疫组化法检测乳腺癌组织ER、PR、Her-2的表达,了解T淋巴细胞亚群在乳腺癌术后患者放疗前、后的变化与ER、PR、Her-2表达之间的相关性。结果 ER受体的阳性表达与Th细胞密切相关(P=0.000),其在Th细胞于放疗前、后无明显变化组的阳性表达率显著高于在放疗后Th细胞较放疗前降低且低于正常值组的表达率;PR受体的阳性表达与Th细胞密切相关(P=0.014),其在Th细胞于放疗前、后无明显变化组的阳性表达率显著高于在放疗后Th细胞较放疗前降低且低于正常值组的表达率。ER、PR受体和Her-2的表达与放疗前、后的总T细胞、Ts细胞、CD4+/CD8+比值和NK细胞等的变化均无关(P>0.05)。结论 ER受体或PR受体阴性表达的患者,术后放疗可引起免疫功能较放疗前降低,预后可能较差。     

EGR1在乳腺癌组织中的表达及临床意义

目的:探究早期生长反应基因1（early growth response 1,EGR1）在乳腺癌组织中的表达及临床意义。方法:下载并提取TCGA数据库中EGR1的表达数据;购买乳腺癌组织芯片（HBreD090CS01）,并采用免疫组织化学法检测EGR1在乳腺癌中的表达;利用Kaplan-Meier Plotter数据库分析EGR1在乳腺癌中的预后价值;利用TIMER数据库分析乳腺癌中EGR1表达与免疫细胞浸润的关系。结果:相较于癌旁组织,EGR1 mRNA在乳腺癌中的表达显著下调（P＜0.001）;EGR1蛋白在45例乳腺癌组织中有18例呈低表达,27例呈高表达,在45例癌旁组织中有9例呈低表达,36例呈高表达,差异具有统计学意义（P=0.038）;EGR1表达与乳腺癌分化程度相关（P=0.003）,与患者年龄、肿瘤大小、N分期、临床分期等无关（P＞0.05）;EGR1低表达的乳腺癌患者,其无复发生存率显著低于EGR1高表达的患者（P＜0.001）,但EGR1表达与乳腺癌患者的总生存率无明显相关性（P＞0.05）;乳腺癌组织中EGR1表达与B细胞浸润水平呈负相关（P＜0.001）,与CD8+T细胞、CD4+T细胞、巨噬细胞、中性粒细胞、树突状细胞浸润水平呈正相关（P＜0.001）。结论:EGR1在乳腺癌组织中的表达较癌旁组织低,且与肿瘤分化程度、预后及免疫浸润有关,有望成为乳腺癌预后评估的分子标志物及治疗的潜在靶点。     

Predicting aggressive outcome in T1N0M0 breast cancer

Despite the excellent overall prognosis, unpredictable breast cancer recurrences and deaths also occur among T1N0M0 patients. We have evaluated clinically applicable methods for identifying aggressive outcome in T1N0M0 breast cancer. The material is based on aggressive T1N0M0 invasive ductal and lobular carcinomas diagnosed in Turku University Hospital and Jyv?skyl? Central Hospital, Finland, during 1987-1997. We studied all the T1N0M0 breast cancers that had led to recurrency or death (n=21, 95% T1cN0M0) during the follow-up period (4-14 years). The study is based on statistical analyses of matched case-control data in which the prognostic factors of each individual patient with aggressive disease were compared with control patients (n=45) individually matched by tumour size, age at diagnosis, histological type of tumour and length of follow-up. The cancer cases were examined for clinically applicable conventional and immunohistochemical pathologic prognostic factors. High Ki-67 immunopositivity was the strongest prognosticator of breast cancer death or recurrence in T1N0M0 breast cancer. Also, high p53 immunopositivity, low oestrogen receptor immunopositivity and Her-2/neu oncogene amplification by chromogen in situ hybridisation were reliable indicators of unfavourable outcome. Our statistical methods also allowed us to determine for the present material a range of clinical significance for each immunohistochemical prognostic feature with the associated relative risk for breast cancer death and recurrence. The paper suggests guidelines for predicting aggressive outcome in T1N0M0 breast cancer.     

Inhibition of breast cancer cell growth in vitro by a tyrosine kinase inhibitor.

Human breast cancer cell proliferation is regulated by growth factors that bind to receptors with intrinsic tyrosine kinase (TK) activity, including the epidermal growth factor (EGF) receptor. To determine whether inhibition of receptor TK activity inhibits tumor growth, we studied the effects of a tyrosine kinase inhibitor, RG-13022, on cultured human breast cancer cells. RG-13022 represents a class of compounds which have been shown to inhibit preferentially the TK activity of the EGF receptor in a cell-free system and also to inhibit EGF-stimulated growth of cultured cells. RG-13022 significantly inhibited EGF-stimulated autophosphorylation of its receptor in two breast cancer cell lines that have abundant, although not amplified, EGF receptor content (MDA-231 and T47D). RG-13022 also inhibited EGF-stimulated DNA synthesis and proliferation of T47D and MCF-7 breast cancer cells in a reversible and dose-dependent manner. Inhibition was observed at 0.1 microM, and it was maximal at 10 microM. The effect was rapid (within 3 h), persisted for 18 h, and was partially reversed by 24 h at 1 microM. At 5 microM, inhibition persisted for more than 50 h. Inhibitory effects were also observed in a panel of estrogen receptor-positive and estrogen receptor-negative breast cancer cell lines. RG-13022 inhibited not only EGF-induced growth but also growth stimulated by insulin, insulin-like growth factor I, insulin-like growth factor II, or transforming growth factor alpha. RG-13022 also totally blocked estrogen-stimulated phosphorylation of the EGF receptor, as well as estrogen-induced cell proliferation, suggesting that functioning TK pathways are required for estrogen action. The TK inhibitor RG-13022 is a potent inhibitor of hormonally regulated growth of human breast cancer. Tyrosine kinase inhibitors have the potential of providing a new strategy for the "endocrine therapy" of breast cancer.