Overexpression of matrix-metalloproteinase-9 in human breast cancer: a potential favourable indicator in node-negative patients

Matrix metalloprotease-9 (MMP-9; 92 kDa type IV collaganase, gelatinase B) is regarded as, important for degradation of the basement membrane and extracellular matrix during cancer invasion and other tissue-remodelling events. In this study we evaluate the prognostic value of MMP-9, by immunoperoxidase staining in a series of 210 breast cancer tissues. The results were quantitated using the HSCORE system, which consider both staining intensity and the percentage of cells stained at given intensities. MMP-9 status was compared with the concentration of cytosolic Cathepsin-D and with other established prognostic factors, in terms of disease free survival and overall survival. The median follow-up period was 62 months. MMP-9 staining was observed primarily in cancer cells, and to a lesser degree in surrounding stromal cells. MMP-9 expression was not detected in normal breast tissue. Levels of MMP-9 expression below the cut-off point were more frequently observed in larger (P = 0.014), invasive ductal histologic (P = 0.037), progesterone receptor (PR)-negative and PR-strong positive tumours (P< 0.001), as well as samples belonging to patients with stage III-IV disease (P = 0.009) and age 45-55 years (P = 0.011). In univariate analysis, node-negative breast cancer patients with tumors positive for MMP-9 had a considerable reduction in risk for relapse (RR = 0.45;P = 0.039) or death (RR = 0.32;P = 0.009). Multivariate analysis indicated that MMP-9 status was an independent favourable predictor of OS (RR = 0.47;P = 0.034) in node-negative but not in node-positive patients. Our results suggest that MMP-9 may be an independent favourable prognostic factor in node-negative breast cancer patients. The overexpression of MMP-9 in breast cancer may be also used as a marker to subdivide node negative breast cancer patients in order to determine the optimal treatment modality.     

MMP-9、BSP与LRP在乳腺癌组织中表达的研究

目的：探讨基质金属蛋白酶-9（MMP-9）、骨唾液蛋白（BSP）与肺耐药蛋白（LRP）在正常乳腺组织及乳腺癌组织中的表达状况。方法：应用免疫组化SP法检测60例正常乳腺组织及60例乳腺癌组织中MMP-9、BSP与LRP的表达。结果：乳腺癌组织中MMP-9、BSP与LRP 的阳性表达率分别为75.0％、78.3％、68.3％。正常乳腺组织中MMP-9、BSP与LRP的阳性表达率分别为21.7％、15.0％、10.0％。二者的差异有显著的统计学意义（ P＜0.05）。乳腺癌组织中MMP-9、BSP与LRP的表达呈正相关。结论：MMP-9、BSP与LRP在乳腺癌组织中高表达,与乳腺癌的浸润和转移相关。     

三阴性乳腺癌中MMP-9的表达及意义

目的:探讨三阴性乳腺癌(TNBC)中基质金属蛋白酶-9(MMP-9)的表达及意义.方法:收集2012年1月至2013年12月乳腺浸润性导管癌共158例,应用免疫组织化学及荧光原位杂交技术将乳腺癌分为65例TNBC、93例非TNBC.免疫组织化学检测MMP-9的表达情况,明确MMP-9在二组间表达的区别.分析MMP-9在TNBC中与临床病理特征关系,并随访65例TNBC患者3年复发或转移率,探讨MMP-9表达与3年复发转移率间的关系.结果:TNBC中MMP-9表达明显高于非TNBC(P<0.05);TNBC中MMP-9表达与年龄和肿瘤大小无关(P>0.05),与TNM分期、组织学分级、淋巴结状态及脉管浸润有关,差异具有统计学意义(P<0.05);MMP-9阳性的TNBC患者3年复发转移率高达76.09%,明显高于MMP-9阴性组47.37%,差异具有统计学意义(P<0.05).结论:TNBC中MMP-9表达明显高于非TNBC,MMP-9与TNBC浸润转移有关,对预测3年复发转移有明显意义.     

Zymographic detection and clinical correlations of MMP-2 and MMP-9 in breast cancer sera

Matrix metalloproteinases, in particular the gelatinases MMP-2 and MMP-9, have received great attention in recent years as putative tumour markers for clinical applications. The main reason for the observed interest is their easy detection in body fluids. Moreover, recent evidence has shown multiple functions of MMPs, rather than simply degrading ECM, which include the mobilisation of growth factors and processing of surface molecules. Several authors have reported increased levels of MMPs in a number of cancers, but clinical correlations in breast cancer are still fragmentary. Thus, the aim of the present research was to investigate the activity levels of circulating gelatinases in the sera of breast cancer patients by means of zymographic analysis, and correlate data with clinicopathological parameters. In all, 80 patients and 22 healthy volunteers were involved in this study. Sera were obtained prior to surgery. The clinical variables were: grading of tumours, tumour size, lymph node involvement, tumour staging, oestrogen and progesterone receptor levels (76 out of 80 cases), and c-erbB-2 levels (46 cases). The densitometric measures of MMP-2 and MMP-9 activity levels indicated that the average values of both gelatinase activities were significantly higher in breast cancers than in control sera (P<0.0001). In addition, our analysis showed for the first time that elevated activity levels of both gelatinases correlated only with c-erbB-2 overexpression (P=0.0273 for MMP-2 and P=0.0075 for MMP-9). An inverse correlation was observed with regard to oestrogen receptor expression (P=0.0075 for MMP-2 and P=0.0273 for MMP-9). Moreover, a borderline inverse correlation was observed between the activity levels of both enzymes and nuclear grade (P=0.0511 for MMP-2 and P=0.0794 for MMP-9). In conclusion, the present data suggest that serum measures of MMP's activity may have diagnostic value for discriminating subgroups of breast cancer patients and support the hypothesis that ERBB2 amplification and/or overexpression enhance signalling pathways that may lead to increased production of gelatinases in c-erbB-2 positive breast cancers with higher metastatic potentialities.     

Analysis of the Relationship between Expressions of TF and MMP-9 and Prognosis of Breast Cancer Patients

OBJECTIVE To investigate expression of the tissue factor (TF)and matrix metalloproteinase-9(MMP-9)in breast cancers, and to assess their expression in relation to possible prognostic significance. METHODS The expression of TF and MMP-9 in 71 breast cancer specimens were determined by EnVision immunohistochemistry, and the positive expressions related to the patient clinical outcome. RESULTS Positive rates of TF and MMP-9 staining were respectively 43.7%and 42.3%.K-M monofactorial analysis showed that the 5-year survival rate of the patients with a positive expression of TF and MMP-9 was lower than those with negative expression(P<0.05).However,the COX multifactorial analysis indicated that TNM staging and lymph node metastasis were the prognostic factors for breast cancer patients,and that TF and MMP-9 could not be used as the independent prognostic factors(P >0.05). CONCLUSION The positive rates of TF and MMP-9 were considerably high in breast cancers,which could provide useful information for patient prognosis.     

MMP-2、MMP-9在乳腺癌中的表达及其意义

目的 检测基质金属蛋白酶(MMP-2和MMP-9)与乳腺癌的淋巴结转移、病理分级、患者年龄及肿瘤大小之间的可能关系,及对术后的影响.方法 65例乳腺癌蜡块,采用免疫组化S-P法染色,以细胞浆中有棕色颗粒计数分为:阴性;弱阳性;中度阳性及强阳性.结合随访材料进行统计分析.结果 MMP-2和 MMP-9的表达与乳腺癌患者的年龄及肿瘤大小无关;MMP-2与淋巴结转移有关,无淋巴结转移者,MMP-2表达较低;有淋巴结转移者,MMP-2表达较高.MMP-9的表达与淋巴结转移情况则无关.MMP-2和MMP-9的表达与病理分级相关.二者均影响预后,MMP-2低表达组术后10年生存率为100%,MMP-2高表达组术后1、3、5及10年生存率分别为100%、86.11%、75.0%、66.67%,两组间有显著性差异(P《0.01);MMP-9低表达组术后10年生存率为100%,MMP-9高表达组术后1、3、5及10年生存率分别为100%、86.49%、72.97%、64.86%,两组间有显著性差异(P《0.01).结论 MMP-2和MMP-9的表达与乳腺癌病理学分级有关;MMP-2与淋巴结转移有关,而MMP-9与淋巴结转移无关.二者高表达生存率均降低,影响预后.     

VEGF、MMP-9在肺癌组织中表达的意义

目的:初步探讨肺癌组织VEGF和MMP-9之间的关系及VEGF影响肺癌浸润转移的作用机制。方法:利用45例肺癌标本行免疫组织化学染色,进行VEGF、MMP-9、iMVD的检测,并结合临床病理参数,综合分析肺癌组织中VEGF、MMP-9的表达与肿瘤类型、大小、淋巴结转移、临床分期、iMVD间的关系,以及VEGF和MMP-9间的相互关系。结果:VEGF、MMP-9蛋白在肺癌组织中的表达与肿瘤大小、淋巴结转移、肿瘤病理分期以及病理类型无显著相关(P>0·05);VEGF、MMP-9表达阳性组的iMVD显著高于阴性组,具有统计学意义(P<0·05);VEGF与MMP-9在肺癌组织中的表达具有相关性,相关关系密切(C=0·471,P<0·001)。结论:VEGF和MMP-9在肺癌组织中的表达密切相关,且两者表达与iMVD相关,推测VEGF和MMP-9可能共同参与肿瘤微血管的生成,从而促进肺癌的浸润转移。     

乳腺浸润性导管癌中COX-2、MMP-9的表达及临床意义

目的:探讨COX-2和MMP-9蛋白在乳腺浸润性导管癌组织中的表达及其相关性.方法:应用免疫组织化学法检测52例乳腺癌组织COX-2和MMP-9蛋白的表达.结果:乳腺浸润性导管癌组织中COX-2阳性表达率为76.9%(40/52),MMP-9阳性表达率为82.7%(43/52),COX-2阳性表达与患者的年龄、肿瘤大小、雌孕激素受体无明显相关性(P＞0.05),而与淋巴结转移、TNM分期、组织学分级有关(P＜0.05);MMP-9阳性表达与患者的年龄、雌孕激素受体无明显相关性(P＞0.05),而与肿瘤大小、淋巴结转移、TNM分期、组织学分级有关(P＜0.05);乳腺浸润性导管癌组织中COX-2、MMP-9蛋白的表达之间存在显著正相关(r=0.448,P＜0.01).结论:乳腺浸润性导管癌组织中COX-2、MMP-9蛋白高表达,且两者具相关性.COX-2蛋白可通过诱导MMP-9蛋白的表达上调,增加乳腺癌细胞的侵袭力,促进乳腺癌浸润、转移.     

宫颈癌组织ILK和MMP9表达临床意义分析

目的：探讨宫颈病变组织中整合素连接激酶（integrin—linkkinase,ILK）、基质金属蛋白酶-9（matrixmetalloprotei—nases,MMP-9）的表达及其临床意义。方法：免疫组化方法检测2010—09-01—2012-06-30我院门诊及住院的60例宫颈鳞状细胞癌、100例宫颈上皮内瘤变（cervicalintraepithelialneopla,CIN）、40例慢性宫颈炎组织中ILK和MMP-9的表达。结果：1）宫颈癌组织中ILK表达率为80．0％（48／60）,明显高于正常宫颈黏膜上皮的0％（0／40）,X2=61．54,P〈0．001;高于CINI～Ⅱ的30．77％（16／52）,X2=27．27,P〈O．001;也高于CINlll的45．83％（22／48）,X2=13．65,P〈O．001。而CINI～Ⅱ与CINHI之间ILK的表达差异无统计学意义,X2=2．404,P=0．121。宫颈癌组织中MMP-9表达率为88．33％（53／60）,明显高于正常宫颈黏膜上皮（0／40）,YX2=75．18,P〈0．001;高于CINI～Ⅱ的28．85％（15／52）,X2=41．33,P〈O．001;但与CINⅢ的79．17％（38／48）相比差异无统计学意义,X2=1．69,P=0．194。MMP-9在CINⅢ的表达明显高于CINI～Ⅱ,X2=25．37,P〈0．001。ILK表达与临床分级、组织学分级和淋巴结转移有关,P值均〈O．05。MMP-9与淋巴结转移有关,P=0．012。ILK和MMP9在宫颈癌组织中表达呈正相关,r=0．429,P〈0．05。结论：ILK、MMP-9在宫颈癌组织中均呈过表达,且两者的过表达可能在宫颈癌浸润转移过程中起重要作用。     

By inhibiting Ras/Raf/ERK and MMP-9, knockdown of EpCAM inhibits breast cancer cell growth and metastasis

Epithelial cell adhesion molecule (EpCAM) is a type I transmembrane protein that is expressed in the majority of normal epithelial tissues and is overexpressed in most epithelial cancers including breast cancer, where it plays an important role in cancer progression. However, the mechanism by which EpCAM promotes the progression of breast cancer is not understood. In this study, we found that EpCAM expression was increased in tumor tissue from breast cancer patients compared to healthy patients. Overexpression of EpCAM in breast cancer cells enhanced tumor cell growth in vitro and increased invasiveness, whereas small interfering RNA-mediated silencing of EpCAM (si-EpCAM) had the opposite effect. EpCAM knockdown led to decreased phosphorylation of Raf and ERK, suppression of malignant behavior of breast cancer cells, and inhibition of the Ras/Raf/ERK signaling pathway. Furthermore, si-EpCAM-mediated invasion and metastasis of breast carcinoma cells required the downregulation of matrix metalloproteinase-9 (MMP-9) through inhibition of this signaling pathway. In conclusion, our data show that knockdown of EpCAM can inhibition breast cancer cell growth and metastasis via inhibition of the Ras/Raf/ERK signaling pathway and MMP-9.     

ADAM15与MMP-2、MMP-9 mRNA在乳腺癌组织中的表达及意义

目的:探讨ADAM15、MMP-2、MMP-9 mRNA在乳腺癌组织中的表达、与临床病理特征的关系及对乳腺癌转移、预后的影响。方法:采用原位杂交技术检测45例乳腺癌和20例乳腺良性病变中ADAM15、MMP-2和MMP-9 mRNA表达情况。分析ADAM15 mRNA表达与乳腺癌临床病理特征的关系并分析ADAM15 mRNA与MMP-2、MMP-9 mRNA表达的相关性。从TCGA数据库获取乳腺癌转录组数据,通过生物信息学分析相关因素与乳腺癌临床病理特征的关系。结果:ADAM15 mRNA在乳腺癌中高表达,并与乳腺癌淋巴结转移和临床分期密切相关,与患者年龄和肿瘤大小无关。乳腺癌中MMP-2和MMP-9 mRNA阳性率均明显升高,且均与淋巴结转移相关。乳腺癌组织中ADAM15和MMP-2、MMP-9的mRNA表达均呈正相关。生物信息学分析表明ADAM15 mRNA在乳腺癌组织中高表达,其高表达与患者预后密切相关。结论:ADAM15 mRNA在乳腺癌中高表达,与淋巴结转移呈正相关并与患者预后呈负相关,ADAM15 mRNA有可能通过上调MMP-2和MMP-9 mRNA的表达而促进乳腺癌转移。     

Momordica cochinchinensis Seed Extracts Suppress Migration and Invasion of Human Breast Cancer ZR-75-30 Cells Via Down-regulating MMP-2 and MMP-9

Objective: Metastases and invasion are the main reasons for oncotherapy failure. Momordica cochinchinensis(Mu Bie Zi in Chinese) had been used for a variety of purposes, and shown anti-cancer action. In this article, wefocused on effects on regulation of breast cancer cell ZR-75-30 metastases and invasion by extracts of Momordicacochinchinensis seeds (ESMCs). Methods: Effect of ESMCs on ZR-75-30 human breast cancer cells proliferationwere evaluated by MTT assay and on invasion and migration by wound-healing and matrigel invasion chamberassays. Expression and protease activity of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, wereanalyzed by Western blotting and gelatin zymography, respectively. Results: ESMC revealed strong growthinhibitory effects on ZR-75-30 cells, and effectively inhibited ZR-75-30 cell invasion in a dose-dependent manner.Western blot and gelatin zymography analysis showed that ESMC significantly inhibited the expression andsecretion of MMP-2 and MMP-9 in ZR-75-30 cells. Conclusions: ESMC has the potential to suppress the migrationand invasion of ZR-75-30 cancer cells, and it might prove to of interest in the development of novel inhibitorsfor breast cancer.     

血清MMP-9对局部晚期乳腺癌新辅助化疗疗效的预测

目的 探讨局部晚期乳腺癌新辅助化疗（NCT）前后血清基质金属蛋白酶-9（MMP-9）水平的变化及其对疗效的预测价值。方法 检测本院2012年7月至2013年6月收治的80例Ⅱ～Ⅲ期乳腺癌患者NCT前后的血清MMP-9水平并评价NCT疗效。64例ER、PR阳性乳腺癌患者采用EC-T方案化疗（表阿霉素90mg/m2静滴,d1;环磷酰胺600mg/m2静滴,d1,21天为1周期,共4个周期;序贯多西他赛80mg/m2静滴,d1,21天为1周期,共4个周期）,16例ER、PR阴性者采用TEC方案化疗（多西他赛75mg/m2静滴,d1;表阿霉素75～85mg/m2静滴,d1;环磷酰胺600mg/m2静滴,d1,21天为1周期,共4个周期）。分析血清MMP-9水平与NCT疗效及临床病理特征的关系。结果 80例患者共完成NCT 290个周期,有效率（RR）为78.8％（63／80）,其中获CR 4例,PR 59例。38例（47.5％）临床分期降低。NCT前血清MMP-9阳性组（＞900U／L）和阴性组（≤900U／L）的RR分别为85.2％（46／54）和65.4％（17／26）,差异有统计学意义（P＜0.05）。血清MMP-9水平与乳腺癌化疗前分期、HER-2、ER/PR状态及病理学反应性分级均有关（P＜0.05）。在化疗有效、化疗前分期Ⅲ期、HER-2+、ER+、PR-/+、病理学反应性分级G1～G3和G4～G5及绝经和未绝经的患者中,化疗2个周期后的血清MMP-9水平与化疗前比较明显降低（P＜0.05）。结论 血清MMP-9水平对乳腺癌NCT疗效具有一定的预测价值,可减少NCT的盲目性,有助于制定有效的治疗方案。     

MMP-9、MMP-2在非霍奇金淋巴瘤病理组织中的表达及意义

目的探讨非霍奇金淋巴瘤(non-Hodgk in's lymphom a,NHL)患者病理组织中MMP-9、MMP-2蛋白的表达及其相互关系,分析其临床及预后的意义。方法采用免疫组织化学S-P法,检测了51例NHL、17例良性增生性淋巴结炎中MMP-2、MMP-9的蛋白表达。结果MMP-9、MMP-2蛋白在NHL和良性增生性淋巴结炎中均表达于胞浆,少见于胞核。MMP-9、MMP-2在NHL中的高表达阳性率分别为84.3%和64.7%,均高于良性增生性淋巴结炎的52.9%和54.3%(P<0.05)。Ⅲ、Ⅳ期NHL患者的MMP-9、MMP-2蛋白的高表达阳性率明显高于Ⅰ、Ⅱ期患者(P<0.05);而MMP-9、MMP-2蛋白的表达与恶性程度、LDH水平、结外病变均无关(P>0.05)。NHL病理组织中MMP-9与MMP-2蛋白的表达呈正相关(rs=0.348,P<0.05)。结论MMP-9、MMP-2蛋白在NHL病理组织中均呈高表达,均与Ann Arbor临床分期有关,可能是预测NHL侵袭转移和预后的相关因素。     

Metabolomics research on potential role for 9‐cis‐retinoic acid in breast cancer progression

Deciphering the molecular networks that discriminate organ‐confined breast cancer from metastatic breast cancer may lead to the identification of critical biomarkers for breast cancer invasion and aggressiveness. Here metabolomics, a global study of metabolites, has been applied to explore the metabolic alterations that characterize breast cancer progression. We profiled a total of 693 metabolites across 87 serum samples related to breast cancer (46 clinically localized and 41 metastatic breast cancer) and 49 normal samples. These unbiased metabolomic profiles were able to distinguish normal individuals, clinically localized and metastatic breast cancer patients. 9‐cis‐Retinoic acid, an isomer of all‐trans retinoic acid, was identified as a differential metabolite that significantly decreased during breast cancer progression to metastasis, and its levels were also reduced in urine samples from biopsy‐positive breast cancer patients relative to biopsy‐negative individuals and in invasive breast cancer cells relative to benign MCF‐10A cells. The addition of exogenous 9‐cis‐retinoic acid to MDA‐MB‐231 cells and knockdown of aldehyde dehydrogenase 1 family member A1, a regulatory enzyme for 9‐cis‐retinoic acid, remarkably impaired cell invasion and migration, presumably through preventing the key regulator cofilin from activation and inhibiting MMP2 and MMP9 expression. Taken together, our study showed the potential inhibitory role for 9‐cis‐retinoic acid in breast cancer progression by attenuating cell invasion and migration.     

MMP-9和TIMP-1在结肠癌组织中的表达及其临床意义

目的 探讨基质金属蛋白酶9（MMP-9）及组织抑制剂1（TIMP-1）在结肠癌中的表达和临床意义。方法 采用RT-PCR、免疫组化、计算机图象分析方法定量检测200例结肠癌及其对应癌旁组织中MMP-9、TIMP-1 基因和蛋白表达水平,并分析其表达与临床病理特征的关系及两者的相关性。结果 结肠癌组织中MMP-9 mRNA的表达上调者141例（70.5％）,癌旁组织为21例（10.5％）,差异有统计学意义（P=0.001）。MMP-9 mRNA表达与结肠癌Dukes分期、淋巴结转移及分化程度相关。结肠癌及其癌旁组织中TIMP-1 mRNA表达上调者分别为104例（52.0％）和121例（60.5％）,差异无统计学意义（P=0.087）。TIMP-1 mRNA表达与结肠癌临床病理特征均无关。MMP-9蛋白在结肠癌组织中的免疫组织化学染色强度为1.79±0.11,高于癌旁组织的1.22±0.05（P=0.002）,MMP-9蛋白表达水平与结肠癌Dukes分期及淋巴结转移相关。TIMP-1蛋白在结肠癌和癌旁组织中表达均较MMP-9蛋白表达弱,其表达与临床病理特征无关。在结肠癌组织中MMP-9与TIMP-1蛋白表达呈负相关 （r=-0.63,P＜0.01）。 结论 MMP-9表达在结肠癌的进展中具有促进作用,而TIMP-1表达则可能具有拮抗作用。