P-gp、MMP-2、C-erbB-2在乳腺癌原发灶和转移淋巴结中的表达对比研究

目的：探讨乳腺癌侵袭转移和多药耐药之间的关系,为治疗方案的个体化提供依据。方法：采用免疫组化方法检测46例乳腺浸润性导管癌患者乳腺原发灶及相应腋淋巴结转移灶中P-gp、MMP-2、c-erbB-2的表达,结合临床表现、病理学指标,分析其相关性。结果：46例原发灶P-gp阳性表达35例（76.1%）,MMP-2阳性表达25例（54.3%）,c-erbB-2高表达18例（39.1%）;相应腋淋巴结转移灶P-gp阳性表达28例（60.9%）,MMP-2阳性表达16例（34.8%）,c-erbB-2高表达16例（34.8%）;P-gp、MMP-2蛋白表达水平与肿块大小、淋巴结转移数目均呈正相关（P〈0.05）,c-erbB-2蛋白表达水平与腋窝淋巴结转移数量呈正相关,与ER、PR表达呈负相关,P-gp阳性表达与MMP-2和c-erbB-2的表达呈正相关（P〈0.05）。结论：肿瘤原发灶与转移灶存在异质性,P-gp、MMP-2、c-erbB-2的表达与乳腺癌的多药耐药和侵袭转移有关,检测上述基因在原发灶与转移灶的表达,为乳腺癌选择个体化的化疗、内分泌治疗及分子靶向治疗提供了分子生物学依据。     

乳腺癌肺耐药蛋白表达及其与C-erbB-2和p53表达的关系

目的　探讨LRP在乳腺癌组织中的表达及其与C erbB 2和 p5 3表达的相关性。方法　采用免疫组化LSAB法检测 15 2例乳腺癌组织和 40例正常乳腺组织的LRP、C erbB 2和 p5 3蛋白的表达水平 ,并将结果与临床病理因素和预后资料进行分析。结果　在 15 2例乳腺癌组织中LRP、C erbB 2和 p5 3阳性例数分别为 12 2 ( 80 2 % )、 10 2 ( 67 1% )和 69( 45 4% ) ,乳腺对照组织LRP阳性 2 2例 ( 5 5 0 % ) ;乳腺癌组织LRP的表达明显高于对照组 ( χ2 =10 70 9,P <0 0 1) ,并与C erbB 2 ( χ2 =12 44 0 ,P <0 0 1,γ =0 2 9)和 p5 3 ( χ2 =8 5 0 0 ,P <0 0 1,γ =0 2 5 )表达呈正相关 ;在淋巴结转移组LRP阳性 62例 ( 88 6% ) ,无转移组阳性 60例 ( 73 2 % ) ,LRP的表达与淋巴结转移 ( χ2 =5 65 4,P <0 0 5 ,γ =0 19)呈正相关。单因素预后分析显示LRP阳性组生存期明显低于阴性组 ( χ2 =7 0 92 ,P <0 0 1)。结论　C erbB 2和p5 3可能诱导LRP表达上调 ,LRP表达可能与腋淋巴结转移、缩短常规化疗乳腺癌患者生存期有关     

P-gp 和MDR1 在三种乳腺癌细胞中表达差异研究

目的:比较P-gp和MDR1在人乳腺癌敏感细胞(MCF-7/S)和耐药细胞(MCF-7/ADR、MCF-7/TAM)中的表达差异,初步探讨乳腺癌细胞对阿霉素与对三苯氧胺产生耐药机制的区别。方法:采用免疫细胞化学法、流式细胞术检测P-gp,采用实时荧光定量PCR法检测MDR1在三种乳腺癌细胞中的表达情况。结果:在MCF-7/ADR细胞中P-gp和MDR1均呈高表达,阳性表达率与MCF-7/S细胞比较,有统计学意义(P<0.01)。在MCF-7/TAM细胞中P-gp、MDR1均呈低表达,与MCF-7/S细胞比较,无统计学意义(P>0.05)。结论:P-gp和MDR1的高表达是乳腺癌细胞对阿霉素产生耐药的主要机制,而并非是乳腺癌细胞对三苯氧胺产生耐药的机制。     

抗转移癌基因在乳腺癌中的表达及其与淋巴结转移和预后的关系

观察 nm2 3 在乳腺癌中的表达与淋巴结转移和预后的关系。方法 :应用免疫组化 L SAB法检测 15 2例乳腺癌 nm2 3、ER、 PR、 P5 3和 PCNA的表达水平 ,比较 nm2 3 的表达对淋巴结转移和预后的影响。结果 :(1)在淋巴结转移组 nm2 3表达例数 32例 (4 5 .7% ) ,无转移组表达 40例 (4 8.8% ) ,两者差异无显著性 (P>0 .0 5 ) ;(2 )在 PCNA高指数表达组 nm2 3 阳性 42例 (6 0 .9% ) ,低指数表达组 nm2 3阳性 30例 (36 .1% ) ,前者明显高于后者 (χ2 =9.2 39,P<0 .0 1) ,且 PCNA与nm2 3 两因素呈正相关关系 (γs=0 .30 4,P<0 .0 1) ;(3)临床病理多因素比例风险回归分析显示淋巴结转移是影响患者生存的主要因素 ,而 nm2 3 对患者生存期无影响。结论 :nm2 3在乳腺癌中的表达既不能预测淋巴结转移 ,也不能提示较长的生存期 ,可能与肿瘤细胞的活跃增生有关。     

Claudin5、MMP-13在食管鳞癌中的表达及临床意义

目的研究Claudin5和MMP-13蛋白在食管鳞癌组织中表达及其临床意义。方法应用免疫组化技术检测90例食管鳞癌组织、28例正常食管黏膜组织中Claudin5和MMP-13蛋白的表达情况。结果食管鳞癌组织中Claudin5和MMP-13蛋白的阳性表达率分别为85.56%(77/90)和82.22%(74/90),而在正常食管黏膜组织中Claudin5和MMP-13蛋白的阳性表达率分别为25.00%(7/28)和21.43%(6/28),两者差异有统计学意义(P0.05)。MMP-13的蛋白表达在有淋巴结转移的病人中明显高于没有淋巴结转移的病人;临床分期Ⅲ+Ⅳ期组的MMP-13蛋白的表达显著高于Ⅰ期、Ⅱ期组(P0.05)。MMP-13和claudin5在食管鳞癌中表达呈正相关(r=0.6356 P0.01)。结论 MMP-13和claudin5在食管鳞癌的发生、发展过程中可能发挥重要作用,MMP-13可能作为食管鳞癌临床判断预后的又一重要指标。     

CD44V6和MMP-2在甲状腺肿瘤中的表达及其临床意义

目的:研究黏附分子CD44V6和基质金属蛋白酶-2(MMP-2)在甲状腺癌中的表达、相互关系及其与甲状腺癌侵袭转移的相关性.方法:采用SP免疫组化法检测114例甲状腺肿瘤组织中CD44V6和MMP-2的表达.结果:CD44V6和MMP-2在35例甲状腺乳头状癌(Papillary Thyroid Carcinoma,PTC)中阳性表达率分别为70.6%和73.5%,在41例甲状腺滤泡癌(Follicular Thyroid Carcinomas,FTC)中阳性表达率分别为70.7%和75.6%,均高于甲状腺腺瘤和结节性甲状腺肿组织中的表达,差异具有统计学意义(P＜0.05).在甲状腺癌组织中MMP-2和CD44V6的表达具有显著相关性(r=0.4828,P＜0.001).且两者表达与甲状腺癌的临床分期及有、无淋巴结转移显著相关.结论:CD44V6和MMP-2的表达与甲状腺癌分化程度、浸润和转移关系密切.CD44V6和MMP-2检测对甲状腺癌的诊断、分化程度、转移趋势和预后评估具有重要参考价值,是甲状腺癌侵袭、转移和预后判断的分子标志物.     

MMP-2、Galectin-3、Cx43与胃癌侵袭和转移的关系

目的:通过检测胃癌组织和癌旁组织中MMP-2、Galectin-3、Cx43的表达情况,研究它们与胃癌发生、发展、侵袭及转移的关系。方法:应用免疫组织化学(SP法)法检测MMP-2、Galectin-3、Cx43在胃癌组织和癌旁组织中的表达情况。结果:MMP-2、Galectin-3在胃癌组织中的阳性表达均明显高于胃癌癌旁组织(P0.05),并且均与浸润深度、淋巴结转移情况和临床分期明显相关(P0.05);MMP-2与胃癌的分化程度有关,而Galectin-3与胃癌的分化程度无关;Cx43在20例胃癌癌旁组织中的阳性表达率达100%,在胃癌组织中阳性表达率为39.7%,在癌旁组织中的阳性表达明显高于胃癌组织,具有统计学意义(P0.05),Cx43的表达在胃癌的分化程度、浸润深度、淋巴结转移情况及临床分期方面存在显著差异,具有统计学意义(P0.05);MMP-2阳性表达与Cx43阳性表达呈负相关(P=0.02,r=-0.292),而与Galectin-3阳性表达均为正相关(P=0.003,r=0.344)。结论:在胃癌的发生、发展过程中Galectin-3与MMP-2有促进作用,而Cx43有抑制作用,三者在胃癌的侵袭和转移中均发挥重要作用。     

c-Met在人乳腺癌组织中的表达及其与明胶酶的关系

目的研究c—Met蛋白在人乳腺癌组织中的表达及其临床意义,探讨其与乳腺癌明胶酶（MMP-2和MMP-9）关系。方法应用免疫组织化学方法检测86例乳腺癌组织c—Met蛋白的表达情况,分析它们与患者临床病理特征和预后的关系;使用siRNA技术特异性下调乳腺癌细胞内源性c—Met后,westernblot方法检测乳腺癌细胞MMP-2和MMP-9表达水平。结果人乳腺癌c—Met蛋白表达阳性率为58．1％,其表达与肿瘤淋巴结转移和临床分期均呈显著正相关（P〈O．01）,与患者总生存期和无复发生存期均呈负相关（P〈O．01）;相关性分析显示：乳腺癌c—Met和MMP-2及MMP-9表达均呈显著正相关（r=0．314和0．322,P〈O．01）;使用siRNA特异性下调乳腺癌MDA—M13-231细胞c—Met表达后,MMP-2和MMP-9表达也显著降低。结论乳腺癌c—Met表达状况与侵袭转移密切相关,其功能可能是通过调控MMP-2和MMP-9表达而发挥作用。     

MMP-2、MMP-7、MMP-9、TIMP-1及TIMP-2在乳腺癌组织中的表达及意义

目的:探讨基质金属蛋白酶及其抑制剂在乳腺癌组织中的表达及其与肿瘤浸润转移的关系,为乳腺癌的临床治疗及预后预测提供基础。方法:选择我院2012年5月至2014年5月收治的乳腺癌患者80例,对所选病例的乳腺癌组织、癌旁组织及正常乳腺组织样本进行检测。观察并比较不同乳腺组织中MMP-2,MMP-7、MMP-9、TIMP-1及TIMP-2 m RNA的表达水平。结果:与正常乳腺组织相比较,乳腺癌组织和癌旁组织中MMP-2、MMP-7、MMP-9,TIMP-1及TIMP-2 m RNA的表达显著增加,差异具有统计学意义(P0.05)。乳腺癌组织中MMP-2、MMP-7、MMP-9、TIMP-1及TIMP-2 m RNA的表达显著高于癌旁组织和正常组织,差异具有统计学意义(P0.05)。随着肿瘤范围扩大,MMP-2、MMP-7和MMP-9 m RNA的表达水平显著增加(P0.05),而TIMP-1和TIMP-2 m RNA表达无显著变化(P0.05)。随着淋巴结转移进展,MMP-2、MMP-7和MMP-9 m RNA的表达显著增加(P0.05),而TIMP-1和TIMP-2 m RNA无显著变化(P0.05)。结论:MMP-2、MMP-7、MMP-9、TIMP-1和TIMP-2的m RNA在乳腺癌组织中呈高表达,这可能与乳腺癌的发生和发展有关,而MMP-2、MMP-7和MMP-9可能有助于预测乳腺癌的侵袭行为。     

C-erBb-2、MMP-9 在结直肠癌中的表达及相关意义

目的:探讨C-erBb-2、基质金属蛋白酶-9(MMP-9)在结直肠癌中的表达与其浸润转移的关系,寻找浸润转移的客观指标及可能相关机制。方法:用免疫组化法测定80例结直肠癌组织中C-erBb-2和MMP-9的表达情况。结果:①C-erBb-2、MMP-9在结直肠癌组织中阳性表达率60.0%,(48/80)和56.2%(45/80)。②在有淋巴结转移的结直肠癌组织中C-erBb-2、MMP-9阳性率分别为75.0%(30/40)、72.5%(29/40),高于无淋巴结转移的结直肠癌组织中的C-erBb-2、MMP-9阳性率45%(18/30)、40.0%(16/40),(P<0.05);③二者共同阳性者淋巴结转移率为78.1%(25/32)高于二者均阴性者淋巴结转移率26.3%(5/19),(P<0.05),④C-erBb-2、MMP-9联合表达存在意义,且为正相关(r=0.257,P<0.05)。⑤C-erBb-2、MMP-9的表达与肿瘤的大小、性别、年龄、浸润层次等因素无关,但与肿瘤分期、淋巴结转移、分化有关。结论:C-erBb-2、MMP-9表达与淋巴结转移密切相关,二者在有淋巴结转移的结直肠癌组织中表达可能存在协同作用,可作为临床检测淋巴结转移的重要指标之一。     

Expression of matrix metalloproteinase (MMP-2) and extracellular matrix metalloproteinases inducer (EMMPRIN) in benign and advanced breast cancer tissue samples

Tumor cell derived matrix metalloproteinases are a family of enzymes associated with the tumor invasion and metastasis. Extracellular matrix metalloproteinases inducer (EMMPRIN) stimulates synthesis of gelatinase A (MMP-2) in peritoneal fibroblasts. In the present study the role of MMP-2 and EMMPRIN in the progression of breast cancer has been investigated. Gelatinase-A and EMMPRIN were analyzed in benign as well as in stage II and stage III breast cancer tissue samples by gelatin zymography assay, immunoprecipation analysis and Western blot analysis with a monoclonal primary antibody specific for EMMPRIN. Our results showed over expression of EMMPRIN in advanced stages of breast cancer tissues compared with benign tumor tissue samples. The expression of MMP-2, the active and latent forms of the enzyme increased with tumor progression from Stage II to Stage III of breast cancer and it was not expressed in benign tissues. The expression MMP-2 correlates with tumor progression. This observation obviously indicates that EMMPRIN and MMP-2 are the major determinants of malignancy in cancers.     

MMP-9 and MMP-2 gelatinases and TIMP-1 and TIMP-2 inhibitors in breast cancer: correlations with prognostic factors

The goal of our study was to analyse the prognostic values for some matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in breast cancer. We evaluated the activity and the expression levels of MMP-9, MMP-2, TIMP-1 and TIMP-2 in malignant versus benign fresh breast tumor extracts. For this purpose, gelatinzymography, immunoblotting and ELISA were used to analyse the activity and expression of MMPs and TIMPs. We found that MMP-9 expression level and activity are increased in malignant tumors. In addition, MMP-9/TIMP-1 and MMP-2/TIMP-2 ratio values obtained by us were significantly different in malignant tumors compared to benign tumors. We suggest that the abnormal MMP-9/TIMP-1 balance plays a role in the configuration of breast invasive carcinoma of no special type and also in tumor growth, while altered MMP-2/TIMP-2 ratio value could be associated with lymph node invasion and used as a prognostic marker in correlation with Nottingham Prognostic Index. Finally, we showed that in malignant tumors high expression of estrogen receptors is associated with enhanced activity of MMP-2 and increased bcl- 2 levels, while high expression of progesterone receptors is correlated with low TIMP-1 protein levels.     

窖蛋白-1、基质金属蛋白酶-2与乳腺肿瘤的侵袭和转移

窖蛋白(caveolin)是分子量为21～24 kD的整合膜蛋白,是胞膜窖(caveolae)的标志性结构分子,其家族成员窖蛋白-1(caveolin-1,Cav-1)参与细胞内许多重要的生命活动.近来研究发现,窖蛋白-1与乳腺上皮细胞转化及乳腺癌的发生密切相关.基质金属蛋白酶(matrix metalloproteinases,MMPs)是基质降解代谢的主要酶类,几乎能降解细胞外基质和基底膜的所有成分,其家族成员明胶酶A(MMP-2)在乳腺癌的浸润和转移过程中起重要作用.新近发现,窖蛋白-1与基质金属蛋白酶-2在胞膜窖中共定位,窖蛋白-1通过抑制基质金属蛋白酶-2的激活来抑制乳腺癌的侵袭和转移,起到肿瘤抑制因子的作用.本文对窖蛋白-1与基质金属蛋白酶-2各自在乳腺肿瘤侵袭转移中的作用及两者关系的研究进行综述.     

Proangiogenic TIE2+/CD31+ macrophages are the predominant population of tumor-associated macrophages infiltrating metastatic lymph nodes

Tumor-associated macrophages (TAMs) accumulate in various cancers and promote tumor angiogenesis and metastasis, and thus may be ideal targets for the clinical diagnosis of tumor metastasis with high specificity. However, there are few specific markers to distinguish between TAMs and normal or inflammatory macrophages. Here, we show that TAMs localize in green fluorescent protein-labeled tumors of metastatic lymph nodes (MLNs) from B16F1 melanoma cells but not in necrotic tumor regions, suggesting that TAMs may promote the growth of tumor cells and the progression of tumor metastasis. Furthermore, we isolated pure populations of TAMs from MLNs and characterized their gene expression signatures compared to peritoneal macrophages (PMs), and found that TAMs significantly overexpress immunosuppressive cytokines such as IL-4, IL-10, and TGF-β as well as proangiogenic factors such as VEGF, TIE2, and CD31. Notably, immunological analysis revealed that TIE2⁺/CD31⁺ macrophages constitute the predominant population of TAMs that infiltrate MLNs, distinct from tissue or inflammatory macrophages. Importantly, these TIE2⁺/CD31⁺ macrophages also heavily infiltrated MLNs from human breast cancer biopsies but not reactive hyperplastic LNs. Thus, TIE2⁺/CD31⁺ macrophages may be a unique histopathological biomarker for detecting metastasis in clinical diagnosis, and a novel and promising target for TAM-specific cancer therapy.     

Resistance to mesenchymal reprogramming sustains clonal propagation in metastatic breast cancer

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Expression of matrix metalloproteinase 8 (MMP-8) and tyrosinase-related protein-1 (TYRP-1) correlates with the absence of metastasis in an isogenic human breast cancer model

The multi-step nature of metastasis poses difficulties in both design and interpretation of experiments to unveil the mechanisms causing the process. In order to facilitate such studies, we have previously derived a pair of breast tumor cell lines that originate from the same breast tumor but which have diametrically opposite metastatic capabilities. In this system, the monoclonal cell line M-4A4 is metastatic to the lungs of athymic mice, whereas clone NM-2C5 is equally tumorigenic but non-metastatic. Here, we report that representational difference analysis (RDA) of cDNA obtained from the two clonal populations revealed an increased expression of tyrosinase-related protein-1 (TYRP-1) and the matrix metalloproteinase-8 (MMP-8) genes in the non-metastatic cell line. RNA and protein analyses in cultured cells and in primary xenograft tissues confirmed that the non-metastatic cell line expresses TYRP-1 and MMP-8 at levels that are at least 20-fold higher than the metastatic counterpart. Other members of the MMP family (MMP-9 and MMP-2) and the tissue inhibitor of metalloproteinase-2 (TIMP-2) were found to be expressed at similar levels in both populations. The effects of MMP-8 and TYRP-1 on in vitro invasion and migration were assessed in cells whose expression of these genes was altered by stable transduction with sense and antisense constructs. Specific down-regulation of MMP-8 in non-metastatic NM-2C5 cells resulted in a 2.5-fold increased capacity to invade through Matrigel. Unlike other members of the matrix metalloproteinase family, MMP-8 has not previously been implicated in the processes of tumorigenesis or metastasis. The successful identification of two proteins that are differentially expressed in these matched clonal cell lines and the tumors that they produce demonstrates the feasibility of using this approach to search for genes that are associated with aberrant differentiation toward metastatic behavior.     

The correlations between HPV16 infection and expressions of c-erbB-2 and bcl-2 in breast carcinoma

Breast carcinoma (BC) is a prevalent malignant tumour occurring in women. Many studies have indicated the role of human papilloma virus type 16 (HPV16) in the pathogenesis of BC; however, the correlations of HPV16 infection with the clinicopathologic features of BC and the expressions of c-erbB-2 and bcl-2 have not yet been elucidated. In this study, HPV16 was detected by amplifying the HPV16 E6 gene by the polymerase chain reaction method, and the expressions of c-erbB-2 and bcl-2 in 40 BCs and 20 normal breast tissue samples, obtained from Shaanxi Province, were examined using the streptavidin-peroxidase method with monoclonal antibodies specific to c-erbB-2 and bcl-2. The infection rate of HPV16 E6 and the positive expression rate of c-erbB-2 were significantly higher in the BCs than in the normal tissues (HPV16 E6: 60% vs. 5%; c-erbB-2: 42.5% vs. 5%, P < 0.05). However, the positive expression rate of bcl-2 was significantly lower in the BCs than in the normal tissues (67.5% vs. 95%, P < 0.05). The infection rate of HPV16 did not correlate with any of the pathological features observed (P > 0.05). HPV16 infection correlated with bcl-2 expression (P = 0.015) but not with c-erbB-2 expression (P = 0.747) in the BCs. Interestingly, HPV16 infection correlated with bcl-2 expression in grade I BCs (P = 0.018) but not in grade II–III BCs (P = 0.633). Our data suggest that HPV16 infection is correlated with bcl-2 expression in BCs.     

乳腺浸润性导管癌组织中CAS蛋白表达的临床病理意义

目的研究乳腺浸润性导管癌组织中细胞凋亡易感蛋白（CAS）表达的临床病理意义。方法选取乳腺浸润性导管癌53例、普通导管增生20例、异型导管增生20例、导管原位癌10例、正常乳腺组织14例,应用免疫组化方法观察CAS蛋白的表达,并探讨CAS与乳腺癌临床病理因素的关系,分析CAS和HER2、ER、PR以及ki-67指数的关系。结果 CAS在正常乳腺、普通导管增生、异型导管增生、导管原位癌、浸润性导管癌中的阳性率逐渐升高,分别为14.3%、25.0%、40.0%、60%、75.5%（P=0.000）,CAS、HER2均与乳腺癌组织学分级、核分裂像、淋巴结转移有关;CAS评分与ki-67指数（r=0.439,P=0.003）和HER2评分（r=0.598,P=0.000）正相关。结论 CAS与乳腺癌的发生、发展、增殖、淋巴结转移有关,可能作为反映乳腺癌生物学行为的肿瘤标记物,CAS蛋白的表达和HER2有一定的相关性。     

Quantitative assessment of distant recurrence risk in early stage breast cancer using a nonlinear combination of pathological,clinical and imaging variables

Use of genomic assays to determine distant recurrence risk in patients with early stage breast cancer has expanded and is now included in the American Joint Committee on Cancer staging manual. Algorithmic alternatives using standard clinical and pathology information may provide equivalent benefit in settings where genomic tests, such as OncotypeDx, are unavailable. We developed an artificial neural network (ANN) model to nonlinearly estimate risk of distant cancer recurrence. In addition to clinical and pathological variables, we enhanced our model using intraoperatively determined global mammographic breast density (MBD) and local breast density (LBD). LBD was measured with optical spectral imaging capable of sensing regional concentrations of tissue constituents. A cohort of 56 ER+ patients with an OncotypeDx score was evaluated. We demonstrated that combining MBD/LBD measurements with clinical and pathological variables improves distant recurrence risk prediction accuracy, with high correlation ( r = 0.98) to the OncotypeDx recurrence score.