MT1-MMP在乳腺癌组织中的表达及临床意义

背景与目的:近年研究表明,膜型基质金属蛋白酶-1(MT1-MMP)在乳腺癌的浸润和转移中有重要作用.然而,有关MT1-MMP在乳腺癌中表达的临床意义的报道极少.本研究探讨MT1-MMP在乳腺癌组织中的表达及其与乳腺癌临床病理特征的关系.方法:采用免疫组化和半定量RT-PCR的方法检测46例乳腺癌手术后标本中MT1-MMP蛋白和mRNA的表达.结果:MT1-MMP蛋自在46例乳腺癌组织中的阳性率为52.2%,其中在Ⅰ期、Ⅱ期和Ⅲ期中的阳性率分别为12.5%、54.8%和85.7%(P＜0.05);在T1、T2和T3三组中的阳性率分别为11.1%、59.4%和80.0%(P＜0.05);在N0、N1和N2三组中的阳性率分别为27.3%、66.7%和100.0%(P＜0.05).MT1-MMP mRNA在所有的乳腺癌标本中均有表达,其相对表达量的均数在Ⅰ、Ⅱ、Ⅲ期乳腺癌分别为0.473、0.695、0.910(P＜0.05);在T1、T2和T3组中分别为0.524、0.715和0.822(P＜0.05);在N0、N1和N2组中分别为0.630、0.702和0.870(P＜0.05).MT1-MMP蛋白和mRNA表达与ER、PR、c-erbB-2的状况无统计学差异(P＜0.05).结论:MT1-MMP蛋白和mRNA在乳腺癌中的表达与肿瘤分期、肿瘤大小和淋巴结转移呈正相关,有可能作为判断乳腺癌浸润转移能力的参考指标之一.     

MT1-MMP和MMP11在肺癌中的表达及其预后价值

目的探讨MT1-MMP和MMP11在肺癌组织中的表达及其与浸润转移和预后的关系。方法采用免疫组化检测47例肺鳞癌和42例肺腺癌组织中MT1-MMP和MMP11蛋白的表达。结果 MT1-MMP蛋白阳性表达率在肺鳞癌和腺癌组织中分别为68.1%(32/47)和64.3%(27/42)(P=0.705),其表达与T分期、淋巴结转移和TNM分期相关(P<0.05)。MMP11蛋白在肺鳞癌和腺癌组织中的阳性表达率分别为61.7%(29/47)和57.1%(24/42)(P=0.662),其表达与淋巴结转移和TNM分期相关(P<0.05)。MT1-MMP和MMP11表达呈正相关(γ=0.332,P=0.001)。Kaplan-Meier生存曲线显示MT1-MMP和MMP11阳性表达组5年生存率均显著低于阴性表达组(P<0.05)。结论MT1-MMP和MMP11的表达与肺癌的浸润转移密切相关,两者的阳性表达均提示肺癌患者的不良预后。     

MT1-MMP的表达与上皮性卵巢癌生物学特性及预后关系的研究

目的　研究膜型基质金属蛋白酶-1(membranetype-1matrixmetalloproteinase,MT1-MMP)蛋白在上皮性卵巢癌组织中的表达及意义。方法　应用免疫组化SP法检测56例上皮性卵巢癌、15例上皮性卵巢良性肿瘤及10例正常卵巢组织中MT1-MMP的表达和微血管密度(microvesseldensity,MVD)。结果　MT1-MMP在卵巢癌组织中的阳性表达率(67. 9% )显著高于卵巢良性肿瘤(4 /15)和正常卵巢组织(1 /10) (P<0. 01)。MT1-MMP的表达水平与临床分期、组织学分级、有无淋巴结转移和MVD密切相关。在单因素生存分析中,MT1-MMP的表达与患者预后不良有关。结论　MT1 -MMP在上皮性卵巢癌中的异常高表达可能在肿瘤的侵袭转移和血管生成中起重要作用,对判断预后有一定的指导意义。     

β-catenin和MT1-MMP在结直肠癌组织中的表达及意义

目的：探讨结直肠癌组织中争catenin的异常表达的意义及与MT1-MMP表达的关系。方法：应用免疫组化SP法检测78例结直肠癌组织和15例癌旁正常黏膜组织中β-catenin、MT1-MMP的表达。结果：15例正常组织中β-catenin均呈胞膜阳性表达,而MT1-MMP呈阴性表达。而在78例结直肠癌组织中β-catenin细胞膜表达缺失,呈胞质或胞核异位表达。β-catenin异常表达率为75．6％,MT1-MMP阳性表达率为70．5％。β-catenin异常表达与结直肠癌的分化程度、转移和Duke分期显著相关,P〈0．05。MT1-MMP表达水平与结直肠癌浸润深度、转移、Duke分期有统计学意义,P〈0．05。β-catenin异常表达与MT1—MMP的高表达在结直肠癌中有显著的正相关性,P〈0．05,r=0．419。结论：β-catenin在胞质或胞核内的异常聚集和MT1-MMP的过度表达,两者共同参与结直肠癌的进展。     

TGF-β1可通过ERK信号通路调节胃癌细胞MMP-2和MMP-9的表达

目的 探讨转化生长因子-β1（TGF-β1）/丝裂原活化的蛋白激酶（MAPK）通路在调节胃癌细胞MMP-2和MMP-9表达中的作用。方法 以MKN45和BGC823胃癌细胞系为研究对象。明胶酶谱法检测TGF-β1对各细胞MMP-2和MMP-9表达的影响,Western蛋白印迹检测在TGF-β1作用下各细胞中MAPK（P38、JNK、ERK）的活化情况。用相应MAPK通路的特异抑制剂对TGF-β1调节各细胞MMP-2和MMP-9的表达进行阻断研究。结果 TGF-β1可上调MKN45和BGC823细胞MMP-2和MMP-9的表达;TGF-β1可诱导BGC823细胞P38、MKN45和BGC823细胞ERK及JNK的活化;ERK特异抑制剂PD98059可明显抑制TGF-β1诱导MKN45和BGC823细胞MMP-2和MMP-9的表达,但P38特异抑制剂SB203580和JNK特异抑制剂SP600125对TGF-β1诱导这两种细胞MMP-2和MMP-9的表达无明显影响。结论 TGF-β1可通过活化ERK信号通路上调MKN45和BGC823胃癌细胞 MMP-2和MMP-9的表达。     

EMMPRIN/CD147和MMP-2在喉癌组织中的表达及其与预后的关系

目的：探讨细胞外基质金属蛋白酶诱导因子（extracellular matrix metalloproteinase inducer，EMMPRIN／CD147）和基质金属蛋白酶-2（matrix metalloproteinases-2，MMP-2）在喉癌的表达，两者的表达关系及其与喉癌浸润、转移和预后的关系。方法：采用免疫组织化学两步法检测48例喉癌组织、10例喉乳头状瘤和15例喉炎中CD147、MMP-2的表达，并与临床病理特征和预后进行对比。结果：喉癌标本中CD147和MMP-2的阳性表达率分别为87．5％（42／48）和75．0％（36／48），明显高于喉炎组26．7％（4／15）和6．7％（1／15），CD147和MMP-2的阳性表达与喉癌临床分期和淋巴结转移情况有关，CD147和MMP-2在喉癌的表达一致率为68．8％，两者表达之间存在正相关（r=0．736，P〈0.001），单因素生存分析显示CD147和MMP-2表达强阳性的喉癌患者生存率低于表达阴性组。结论：CD147和MMP-2与喉癌侵袭与转移有关，CD147与MMP-2的产生有密切的关系，两者可作为喉癌判断临床预后的参考指标。     

EMMPRIN、MT1-MMP及MMP2在人脑胶质瘤中的表达及意义

目的:探讨基质金属蛋白酶诱导因子(EMMPRIN)、膜型基质金属蛋白酶-1(MT1-MMP)及基质金属蛋白酶2(MMP2)在人脑胶质瘤中的表达及基质金属蛋白酶诱导因子(EMMPRIN)的表达与肿瘤生物学行为和预后的关系.方法:用免疫组织化学S-P法检测48例人脑胶质瘤组织和10例正常人脑组织中EMMPRIN、MT1-MMP和MMP2的表达.结果:高度恶性胶质瘤组织(Ⅲ-Ⅳ级)中EMMPRIN、MT1-MMP和MMP2的阳性表达率显著高于低度恶性胶质瘤组织(Ⅰ-Ⅱ级)和正常脑组织;EMMPRIN、MT1-MMP及MMP2在人脑胶质瘤中共同表达,EMMPRIN的表达与MT1-MMP及MMP2的表达呈显著正相关;在单因素生存分析中,EMMPRIN的表达可能与患者预后不良有关.结论: EMMPRIN在恶性胶质瘤组织中高表达,与胶质瘤的进展和侵袭密切相关,可作为胶质瘤恶性表型的有用指标;EMMPRIN在胶质瘤中的阳性表达可能与患者的预后不良有关;EMMPRIN、MT1-MMP及MMP2三者协同作用,可能共同促进脑胶质瘤的侵袭性发展.     

MT1-MMP及MMP2在人脑胶质瘤中的表达及意义

目的探讨膜型基质金属蛋白酶-1在人脑胶质瘤中的表达及其与肿瘤生物学行为和预后的关系。方法用免疫组织化学S-P法检测48例人脑胶质瘤组织和10例正常人脑组织中MT1-MMP和MMP2的表达。结果高级别胶质瘤组织(Ⅲ~Ⅳ)中MT1-MMP和MMP2的阳性表达率显著高于低级别胶质瘤组织(Ⅰ~Ⅱ),且两者的表达呈显著正相关性。在单因素生存分析中,MT1-MMP的表达与患者预后不良有关。结论MT1-MMP在恶性胶质瘤组织中高表达,与胶质瘤的进展和侵袭密切相关,可作为胶质瘤恶性表型的有用指标;MT1-MMP的阳性表达与患者的预后不良有关。MT1-MMP可能与MMP2的活化密切相关,直接或间接的促进脑胶质瘤的侵袭。     

MT1-MMP和TIMP-2在非小细胞肺癌中的表达及与病理特征的相关性

目的：研究NSCLC中MT1-MMP及TIMP-2的表达及与病理类型分型、淋巴结转移和TNM分期关系。方法：采用免疫组化SP法检测MT1-MMP及TIMP-2在80例NSCLC患者和80例癌旁组织中的表达。结果：在NSCLC中MT1-MMP阳性表达率为73.8%,癌旁组织中为52.5%,（P〈0.05）。在NSCLC中TIMP-2的阳性表达率为42.5%,癌旁组织为72.5%,（P〈0.05）。MT1-MMP和TIMP-2的表达与肺癌的病理分类及肿瘤大小无关,而与TNM分期及淋巴结转移有关。MT1-MMP和TIMP-2的表达呈负相关（r=-0.635,P〈0.05）。结论：MT1-MMP和TIMP-2可能参与了NSCLC的发生发展过程,有望成为判断肺癌转移和预后的指标。     

MT1-MMP和TIMP-2在非小细胞肺癌中的表达及与病理特征的相关性

目的 研究NSCLC中MT1-MMP及TIMP-2的表达及与病理类型分型、淋巴结转移和TNM分期关系.方法 采用免疫组化SP法检测MT1-MMP及TIMP-2在80例NSCLC患者和80例癌旁组织中的表达.结果 在NSCLC中MT1-MMP阳性表达率为73.8%,癌旁组织中为52.5%,(P<0.05).在NSCLC中TIMP-2的阳性表达率为42.5%,癌旁组织为72.5%,(P<0.05).MT1-MMP和TIMP-2的表达与肺癌的病理分类及肿瘤大小无关,而与TNM分期及淋巴结转移有关.MT1-MMP和TIMP-2的表达呈负相关(r=-0.635,P<0.05).结论 MT1-MMP和TIMP-2可能参与了NSCLC的发生发展过程,有望成为判断肺癌转移和预后的指标.     

血管内皮生长因子与膜型基质金属蛋白酶-1在口腔鳞状细胞癌表达及意义

[目的]检测血管内皮生长因子（VEGF）和膜型基质金属蛋白酶-1（MT1-MMP）在口腔鳞状细胞癌（OSCC）中的表达,并探讨其意义。[方法]采用免疫组织化学SP法检测VEGF、MT1-MMP在40例OSCC和10例口腔正常黏膜组织中的表达。[结果]VEGF和MT1-MMP在OSCC组织中的阳性表达率分别为67．50％和75．00％,在正常口腔黏膜组织中均阴性表达．差异有显著性（P〈0．001）。VEGF表达与OSCC的临床分期（P〈0．05）及淋巴结转移显著相关（P〈0．011,与肿瘤的分化程度无关。MT1-MMP表达与OSCC的分化程度（P〈0．05）及淋巴结转移显著相关（P〈0．01）,但与肿瘤的临床分期无相关性。VEGF与MT1-MMP表达呈正相关（rs=0．650,P〈0．001）。[结论]VEGF与MT1-MMP在OSCC发生发展中起重要作用,共同促进OSCC的浸润转移。     

Galectin-3 induces cell migration via a calcium-sensitive MAPK/ERK1/2 pathway

The presence and level of circulating galectin-3 (Gal-3), a member of the galectin family, is associated with diverse diseases ranging from heart failure, immune disorders to cancer metastasis and serves as a biomarker of diagnosis and treatment response. However, the mechanisms by which exogenous Gal-3 affects pathobiology events remain elusive. In the current study, we found that exogenous Gal-3 slightly delays, while prolonging tyrosine phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in HeLa cells through a calcium-sensitive and PKC-dependent signaling pathway. The activation was dependent on the sugar-binding properties of Gal-3, since the antagonist lactose could inhibit it. The sugar-binding motif of Gal-3 was required for the activation of ERK1/2. The activation of ERK1/2 was necessary for the initiation and induction of cell migration associated with the phosphorylation of paxillin. All the results presented in this study suggest a novel calcium-sensitive and PKC-dependent pathway through which circulating Gal-3 promotes cell migration and activating the ERK1/2. Taken together, the data depicted here propose a biological function and a target for the diseases'' associated circulating Gal-3.     

Differential Effects of Inhibitory and Stimulatory Anti-HER2 Monoclonal Antibodies on AKT/ERK Signaling Pathways

Objective: Homo- and heterodimerization of the receptor tyrosine kinase HER2 hyperactivate several downstreamsignaling pathways, leading to uncontrolled growth and proliferation of tumor cells. Anti-HER2 monoclonal antibodies(mAbs) may induce different effects on HER2 dimerization and signaling. Methods: The effect of two inhibitory(2A8, 1T0) and one stimulatory (1H9) anti-HER2 mAbs either alone or in combination with trastuzumab was investigatedon AKT and ERK signaling pathways and HER2 degradation in a human breast cancer cell line (BT-474) by Westernblotting. Result: While 1H9 mAb had no significant effect on AKT and ERK signaling pathways, 1T0 and 2A8 mAbsinhibited phosphorylation of both pathways. Combination of 1T0 mAb with trastuzumab resulted in significant synergisticinhibition of both pathways and HER2 degradation, much more potently than the combination of trastuzumab andpertuzumab. Conclusion: Our data indicate that anti-HER2 mAbs may induce different signaling pathways dependingon their effect on tumor cell growth and proliferation. The significant inhibition of ERK and AKT phosphorylation by1T0 alone or particularly in combination with trastuzumab suggests its potential therapeutic application for targetedimmunotherapy of HER2 overexpressing malignancies.     

Dual blockade of EGFR and ERK1/2 phosphorylation potentiates growth inhibition of breast cancer cells

One of the major targets for breast cancer therapy is the epidermal growth factor receptor (EGFR) and related receptors, which signal via different signal transduction pathways including the mitogen-activated protein kinase (MAPK) pathway. This study determined whether there is a correlation between EGFR/HER2 status and MAPK (ERK1/2) phosphorylation in breast cancer cells, and how this affects the response to an inhibitor of the receptors. Expression of EGFR, HER2 and phosphorylated ERK1/2 were measured by immunoblotting in a panel of breast cancer cell lines. Several lines expressed high levels of pERK1/2, with no obvious correlation with the level of EGFR/HER2. The EGFR tyrosine kinase inhibitor PKI166 inhibited growth and induced apoptosis in some cells with high levels of growth factor receptors (MDA-MB-468, SUM149, SKBR3), but was less effective in cells that also had high basal ERK1/2 activity (MDA-MB-231). The combination of an inhibitor of MAPK signalling (U0126) and PKI166 produced significantly more inhibition and apoptosis than either agent alone. This suggests that constitutive activation of the MAPK pathway may bypass inhibition of EGFR/HER2 tyrosine kinases, and lead to insensitivity to agents targeting the receptors. However, inhibiting both EGFR/HER2 and MAPK signalling can result in significant growth inhibition and apoptosis of EGFR-expressing breast cancer cells.     

Integrin alpha(v)beta6-associated ERK2 mediates MMP-9 secretion in colon cancer cells

There is general consensus that matrix metalloproteinases are involved in tumour progression. We show herein that inhibition of integrin alpha(v)beta6 expression in colon cancer cells suppresses MMP-9 secretion. This integrin-mediated event is dependent upon direct binding between the beta6 integrin subunit and extracellular signal-regulated kinase 2. Targetting either beta6 or its interaction with extracellular signal-regulated kinase in order to inhibit matrix metalloproteinase activity may offer a useful therapeutic approach in preventing growth and spread of colon cancer.     

Oestrogen Receptor-Mediated Modulation of the EGFR/MAPK Pathway in Tamoxifen-Resistant MCF-7 Cells

Oestrogen receptor (ER) levels are usually maintained on acquisition of tamoxifen resistance in the clinic, however, tumour re-growth is associated with increased expression of epidermal growth factor receptor (EGFR) and activation of the mitogen activated protein kinase (MAPK) pathway. In the present study we have used the ER down-regulator fulvestrant ('Faslodex') to investigate the influence of the ER on growth of a tamoxifen-resistant (TAM-R) human breast cancer cell line. Expression levels of ER mRNA and protein were equivalent in parental wild-type MCF-7 (WT) and TAM-R cells. Fulvestrant eliminated ER protein expression and inhibited proliferation in both cell lines. The growth inhibitory effects of fulvestrant were associated with a decrease in basal EGFR, c-erbB2 and ERK1/2 activity in TAM-R but not WT cells. ER functionality as determined by oestrogen response element (ERE)-luciferase reporter activity and expression of PgR, pS2 and transforming growth factor alpha (TGF) was significantly reduced in TAM-R compared to WT cells and was further decreased by fulvestrant treatment in both cell lines. Epidermal growth factor (EGF) and TGF significantly increased EGFR/MAPK pathway activity in both cell lines. Ligand-induced EGFR/MAPK activation promoted TAM-R cell growth in both the absence and presence of fulvestrant, whereas no proliferative activity was observed under the same conditions in WT cells. These results suggest that the ER modulates EGFR/MAPK signalling efficiency in TAM-R cells possibly through the regulation of TGF availability. This effect may be overcome by the action of exogenous EGFR ligands, which strengthen EGFR/MAPK signalling activity to generate endocrine-insensitive cell growth.     

MicroRNA-21 suppresses PTEN and hSulf-1 expression and promotes hepatocellular carcinoma progression through AKT/ERK pathways

MicroRNAs (miRNAs) have been believed to associate with malignant progression including cancer cell proliferation, apoptosis, differentiation, angiogenesis, invasion and metastasis. However, the functions of miRNAs are intricate, one miRNA can directly or indirectly target multiple genes and function as oncogene or tumor suppressor gene. In this study, we found that miR-21 inhibits PTEN and human sulfatase-1 (hSulf-1) expression in hepatocellular carcinoma (HCC) cells. The hSulf-1 is a heparin-degrading endosulfatase, which can inhibit the heparin binding growth factor-mediated signaling transduction into cells. Therefore, miR-21-mediated suppression of both hSulf-1 and PTEN led to activation of AKT/ERK pathways and epithelial–mesenchymal transition (EMT) in HCC cells, and finally enhance the activity of HCC cell proliferation and movement and promote HCC xenograft tumor growth in mouse models. These findings may provide candidate targets for prevention and treatment of HCC.     

Cross-talk between extracellular S1P/S1P2 and P42/44 MAPK in bcr/abl positive chronic myeloid leukemia cells

Objective: BCR/ABL oncoprotein-expression is associated with uncontrolled cell growth. Sphingosine kinase 1 (SPK1) regulates the production of sphingosine 1-phosphate (S1P), a key lipid signal molecular in cell proliferation and survival. The objective of this study was to elucidate the roles of S1P and its receptors in bcr/abl positive chronic myeloid leukemia (CML) cells.Methods: The expressions of S1P receptors: S1P1, S1P2 and S1P3 in CML cells were detected by RT-PCR. SPK1 expression, activity and extracellular S1P were determined in ECV304 and HL-60 cells which were transfected with bcr/abl gene. To elucidate the relationship between the BCR/ABL, ERK/MAPK (extracellular signal-regulated kinase/mitogen-activated protein kinase), SPK/S1P and S1P/S1P2 signal pathways, bcr/abl positive CML cell line K562 was treated with STI571, PD98059, N,N-dimethyl sphingosine (DMS) and JTE-013.Results: Retrovirus-mediated overexpression of bcr/abl gene in ECV304 and HL-60 cells resulted in upregulation of the expression, activity of SPK1 and increase of the secretion of S1P, whereas treatment of STI571 and PD98059 decreased the BCR/ABL-induced S1P secretion. Treatment of DMS reduced S1P secretion and P42/44MAPK phosphorylation. S1P2-selective antagonist JTE-013 could also decrease P42/44MAPK phosphorylation. Conclusion: These results suggest that BCR/ABL up-regulates extracellular sphingosine 1-phosphate through sphingosine kinase 1 and there is cross-talk between SPK1/S1P/S1P2 and P42/44MAPK in bcr/abl positive CML cells.     

SIGNIFICANCE AND CORRELATION OF MAPK/ERK2 AND PI3-K IN HUMAN BREAST TUMORIGENESIS

Objective： MAPK （（Mitogen-actived Protein Kinase） and PI3-K （Phosphatidylinositol 3-kinase） pathways have been implicated in the mitogenic pathways regulating cell growth, proliferation, differentiation and transformation and thus involved in tumorigenesis. This study was designed to examined the protein expression, activity and mRNA levels of both ERK and PI3-K in a series of breast tumors and adjacent mammary glands, and to figure out the changes of ERK2 and PI3-K during the dynamic process of breast tumorigenesis. Methods： A series of breast tumors and adjacent mammary glands were collected at surgery, including 37 cases of breast cancer, 6 cases of atypical hyperplasia-breast carcinoma in situ and 15 cases of benign conditions. Western blot, kinase activity assay and RT-PCR were used to detect the protein expression, kinase activity and mRNA level, respectively. Results： The revels of protein, activity and mRNA of ERK2 were elevated during the stages of both initiation and progression. The increasing tendency in breast cancer was equal to atypical hyperplasia -in situ carcinoma, but higher than in benign lesion and adjacent normal mammary gland. PI3-K was activated during the stage of progression of breast cancer. An inverse correlation between the activity of PI3-K and ERK2 in breast cancer was found. Conclusion： Our findings indicate that ERK2 may perform its function during both the stages of breast cancer initiation and breast cancer progression, while PI3-K may exert its effect during the stage of breast cancer progression. Both PI3-k and ERK2 are involved in the tumorigenesis of breast cancer.