MMPs与乳腺癌侵袭转移研究的现状

目的:探讨基质金属蛋白酶(MMPs)在肿瘤侵袭和转移过程中的作用机制及其在乳腺癌临床诊断与治疗中的应用价值.方法:以乳腺癌、侵袭转移和MMPs为关键词,检索近几年PubMed全文.纳入标准:1)肿瘤转移机制的最新研究进展;2)MMPs结构及生物学特性的研究;3)与乳腺癌相关的MMPs的最新研究进展.根据纳入标准,最后纳入分析28篇文献.结果:MMPs不仅降解细胞外基质,还促进或抑制肿瘤血管的新生,以及诱导肿瘤细胞免疫耐受.MMPs在乳腺癌中的特异性高表达,为乳腺癌的早期诊断与治疗提供了一种新思路.结论:MMPs与乳腺癌侵袭转移关系尤为密切,将有望成为乳腺癌临床研究中重要的诊断及判断预后的指标.     

CD44基因变异体对人乳腺癌细胞株MCF-7的侵袭作用

目的 探讨一种新的CD44基因变异体(CD44v17)对人乳腺癌细胞株MCF-7侵袭能力的影响及其机制.方法 以人乳腺癌耐阿霉素细胞株(MCF-7/ADR)cDNA为模板,采用聚合酶链反应(PCR)扩增目的片段,将其T-A克隆后,进行测序;构建CD44v17质粒真核表达载体(pcDNA3.1-CD44v17);应用脂质体转染法将pcDNA3.1-CD44v17转染入MCF-7细胞中,采用逆转录聚合酶链反应(RT-PCR)及明胶酶谱法测定转染细胞基质金属蛋白酶(MMP)-2和MMP-9的表达;Transwell法检测CD44v17对MCF-7细胞的侵袭力;Western blot检测胞外信号调节蛋白激酶(ERK)及磷酸化蛋白激酶(p-ERK)变化.结果 限制性内切酶消化证实,重组载体已正确克隆;DNA序列分析显示,CD44v17包含CD44基因1～4号外显子、16～17号外显子、18号外显子1～205位碱基(GeneBank NO.FJ216964).MCF-7细胞转染peDNA3.1-CD44v17后,CD44 mRNA表达量和蛋白表达率分别为0.92±0.22和(70.0±2.5)%;透明质酸(HA)处理后,MMP-2 mRNA表达量和蛋白活性分别为0.72±0.22和1.14.4-0.12,MMP-9 mRNA表达量和蛋白活性分别为0.85±0.19和1.23±0.25,细胞侵袭能力明显增加,侵袭细胞数目为352±33个/视野,而CD44单抗可以阻断这种作用;CD44单抗和促分裂原活化蛋白激酶(MAPK)通路抑制剂预处理转染细胞后,显著阻断了P-ERK的表达.结论 在MCF-7/ADR细胞中发现CD44v17,并成功克隆和建立pcDNA3.1-CDd4v17;HA与CD44v17受体结合,通过CD44→ras→MAPK信号传导通路调节MMP-2和MMP-9的表达,从而增加MCF-7细胞的侵袭力.     

血清MMP-9在肺癌侵袭和转移中的意义

摘 要：［目的］ 研究基质金属蛋白酶-9(matrix metalloproteinase 9,MMP-9)在肺癌患者侵袭和转移中的意义。［方法］ 收集肺癌患者血清70例,健康体检者血清60例,通过酶联免疫吸附法(ELISA)检测血清样本中MMP-9的表达水平。［结果］ MMP-9在肺癌患者血清中的含量明显高于健康人群;且有淋巴结转移组血清MMP-9水平显著性高于无淋巴结转移组;在不同病理类型肺癌组间MMP-9水平差异无统计学意义。［结论］ MMP-9可以作为临床判断肺癌侵袭和转移的生物学指标。     

基质金属蛋白酶与食管癌侵袭转移

基质金属蛋白酶(MMP)在食管癌侵袭转移中起重要作用。MMP在食管癌的表达有明显增加，其表达水平与食管癌预后具相关性。MMP抑制剂(TIMP)可望成为食管癌临床用药。现综述近年来有关MMP与肿瘤血管生成和食管癌表达、侵袭转移方面的研究进展。     

MMP-9与乳腺癌淋巴结转移的关系

目的 探讨MMP-9与乳腺癌淋巴结转移的关系.方法 采用ELISA法检测血清中MMP-9的含量.结果 有淋巴结转移组的乳腺癌患者血清中MMP-9的含量为(1 073.72±93.33)mg/L,无淋巴结转移组为(734.53±75.36)mg/L,正常对照组为(418.53±77.31)mg/L.有淋巴结转移组的乳腺癌患者血清中MMP-9的含量显著高于无淋巴结转移组,差异具有统计学意义(P<0.05);乳腺癌患者血清中MMP-9的含量均显著高于正常对照组,差异具有统计学意义(P<0.05).结论 MMP-9与乳腺癌的淋巴结转移有关.     

乳腺癌组织中转化生长因子β1的表达及其介导乳腺癌侵袭和转移的机制

目的 探讨转化生长因子β1(TGF-β1)蛋白和mRNA在乳腺痛组织中的表达,及其与明胶酶和明胶酶抑制物的关系.方法 建它组织芯片平台,应用免疫组化SP法检测160例乳腺痛组织TGF-β1、基质金属蛋白酶(MMP)-2、MMP-9、组织金属蛋白酶抑制物(TIMP)-1和TIMP-2蛋白的表达;应用原位分子杂交方法检测乳腺癌组织中TGF-β1 mRNA的表达.结果 160例乳腺癌TGF-β1、MMP-2、MMP-9、TIMP-1和TIMP-2蛋白表达的阳性率分别为73.7%、96.9%、95.0%、87.5%和89.4%,TGF-β1 mRNA表达的阳性率为56.2%.TGF-β1的蛋白表达与腋窝淋巴结转移、TNM分期和c-erbB-2表达有关(P＜0.05,P＜0.05和P＜0.01),TGF-β1的mRNA表达与腋窝淋巴结转移有关(P＜0.05).TGF-β1蛋白表达阳性组中位总生存期(OS)为60个月,中位无复发生存期(RFS)为59个月;TGF-β1蛋白表达阴性组中位OS为61个月,中位RFS为61个月,两组生存率差异无统计学意义(P=0.090),无复发生存率有统计学意义(P=0.023).TGF-β1的蛋白表达与MMP-2和MMP-9的表达均呈正相关(r=0.170,P＜0.05;r=0.221,P＜0.01).结论 TGF-β1的表达与乳腺癌侵袭和转移密切相关,TGF-β1的蛋白产物通过调控MMP-2和MMP-9促进乳腺癌的侵袭和转移.     

乳腺癌组织中基质金属蛋白酶-2、9活性检测的意义

基质金属蛋白酶 -２、９(ｍａｔｒｉｘｍｅｔａｌｌｏｐｒｏｔｅｉｎ -ａｓｅ-２、９,ＭＭＰ-２、９)降解底物是基底膜的主要成分 -Ⅳ胶原 ,使基底膜发生缺损 ,以致肿瘤细胞向周围组织浸润和转移 ,影响肿瘤的预后。为此 ,本文利用酶谱 (ｚｙｍｏｇｒａｐｈｙ)和ＵＶＰ软件检测并分析了３５例乳腺癌中ＭＭＰ -２、９活性 ,旨在探讨其与乳腺癌侵袭、转移的关系。１材料与方法１.１标本３５例手术切除标本 ,均经病理学诊断为乳腺癌 ,其中转移组１１例 (远处淋巴结转移 ) ,浸润组１４例 (管状癌９例、浸润性小叶癌５例 ) ,非浸润组１０例 (导管…     

人参多糖注射液对人乳腺癌MCF-7细胞侵袭迁移能力的影响及机制研究

目的 探讨人参多糖注射液对人乳腺癌细胞株MCF-7侵袭迁移能力的影响。方法 以不同作用浓度(12.5μL/mL、25μL/mL、50μL/mL、100μL/mL)人参多糖注射液处理MCF-7细胞株,以PBS作为空白对照,分别采用Transwell小室侵袭实验、划痕实验及Western blot检测细胞侵袭能力、迁移能力及基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)表达。结果 人参多糖注射液对MCF-7细胞体外侵袭和迁移能力具有显著的抑制作用,并呈现剂量及时间依赖性。人参多糖注射液处理后,MCF-7细胞MMP-2蛋白表达及MMP-9蛋白表达比值显著降低,并呈现剂量及时间依赖性。结论 人参多糖注射液可显著抑制人乳腺癌细胞MCF-7侵袭能力和迁移能力,下调MMP-2及MMP-9蛋白表达是其可能的作用机制。     

大肠癌细胞侵袭转移的PKC调节机制研究

目的　探讨蛋白激酶C(PKC)对大肠癌细胞侵袭转移的调节机制。方法　采用羊膜侵袭培养系统和明胶酶谱分析的方法 ,研究PKC激活剂佛波酯PMA ,对人大肠癌细胞株HT 2 9体外侵袭作用的影响及PKC抑制剂staurosporine(SP)对PMA的拮抗作用 ,研究这种体外的侵袭作用与细胞分泌 72kD的基质金属蛋白酶MMP 2和 92kD的基质金属蛋白酶MMP 9的关系。结果　PMA可显著增强HT 2 9细胞的侵袭性 ,与对照组相比 ,有显著性差异 (P <0 .0 1) ,而SP则可拮抗PMA的这种诱导作用。PMA还可增加HT 2 9细胞分泌MMP 2和MMP 9,而SP则可拮抗PMA的这种诱导作用 ,抑制MMP 2和MMP 9的分泌。结论　PKC可调节大肠癌细胞侵袭转移 ,PKC的激活可诱导大肠癌细胞侵袭性增强和增加MMP 2和MMP 9的分泌 ,PKC的抑制可促进大肠癌细胞侵袭性降低和减少MMP 2和MMP 9的分泌。MMP 2和MMP 9的分泌与肿瘤细胞侵袭性有密切关系 ,PKC可能通过调节MMP 2、MMP 9的分泌来影响肿瘤细胞侵袭和转移特性的     

透明质酸酶与血管内皮生长因子在人乳腺癌侵袭与转移中的作用

目的研究透明质酸酶(H yase)与血管内皮生长因子(VEG F)在人乳腺癌侵袭与转移中的意义。方法应用组织匀浆、ELSA及免疫组化等方法测定H yase和VEG F在人乳腺癌中的表达并对其病理特征进行比较。结果乳腺癌组织学分级Ⅰ级、Ⅱ级、Ⅲ级组H yase的均值分别为(3.99±1.91)m U/g、(8.01±2.59)m U/g和(12.00±3.96)m U/g,Ⅲ级和Ⅱ级组H yase表达明显高于Ⅰ级组,Ⅲ级组明显高于Ⅱ级组,其差异具有显著的统计学意义(P<0.05)。乳腺癌无淋巴结转移组和淋巴结转移组H yase的均值分别为(5.25±2.84)m U/g和(8.21±3.26)m U/g,乳腺癌淋巴结转移组H yase表达明显高于无淋巴结转移组。VEG F阳性率在淋巴结转移组为82.6%,明显高于无淋巴结转移组33.3%(P<0.05)。VEG F阳性组和阴性组H yase为(8.39±3.26)m U/g和(5.16±2.58)m U/g,二者间具有显著性统计学差异(P<0.05)。结论H yase和VEG F一样,与人乳腺癌侵袭与转移相关。     

黄芩素与乳腺癌侵袭转移关系的研究新进展

乳腺癌（breast cancer,BC）近年来有发病年轻及发病率上升的趋势,转移是晚期患者的最终阶段和导致乳腺癌患者死亡的主要原因。由于传统术后放化疗普遍存在严重的不良反应,因此从天然物质中寻找毒副反应小、安全有效的抗乳腺癌药物成为近些年的研究热点。黄芩素是一个广泛使用的中草药,具有广泛的抗肿瘤活性。本文主要简述黄芩素与乳腺癌侵袭转移关系的研究,综合阐述其对乳腺癌转移作用机制的影响。     

Lipocalin 2 promotes inflammatory breast cancer tumorigenesis and skin invasion

Inflammatory breast cancer (IBC) is an aggressive form of primary breast cancer characterized by rapid onset and high risk of metastasis and poor clinical outcomes. The biological basis for the aggressiveness of IBC is still not well understood and no IBC‐specific targeted therapies exist. In this study, we report that lipocalin 2 (LCN2), a small secreted glycoprotein belonging to the lipocalin superfamily, is expressed at significantly higher levels in IBC vs non‐IBC tumors, independently of molecular subtype. LCN2 levels were also significantly higher in IBC cell lines and in their culture media than in non‐IBC cell lines. High expression was associated with poor‐prognosis features and shorter overall survival in IBC patients. Depletion of LCN2 in IBC cell lines reduced colony formation, migration, and cancer stem cell populations in vitro and inhibited tumor growth, skin invasion, and brain metastasis in mouse models of IBC. Analysis of our proteomics data showed reduced expression of proteins involved in cell cycle and DNA repair in LCN2‐silenced IBC cells. Our findings support that LCN2 promotes IBC tumor aggressiveness and offer a new potential therapeutic target for IBC.     

人脂联素重组体体外对乳腺癌细胞侵袭和转移的抑制

目的：研究人脂联素重组体对人乳腺癌MDA—MB-231细胞体外侵袭能力的影响。方法：将MDA—MB-231细胞分设对照组和脂联素重组体加药（2．5、5、0、10、20、30μg／ml）组，药物作用一定时间后，分别以MTT法、Transwell小室实验，Chamber小室实验，黏附实验检测脂联素重组体对乳腺癌细胞增殖、侵袭、迁移和黏附能力的影响，以明胶酶谱法检测其对肿瘤细胞分泌MMP-2和MMP-9的影响。结果：脂联素重组体质量浓度高于5．0μg／ml时，对MDA—MB-231细胞生长具有明显的抑制作用（P〈0．01）；能明显地降低肿瘤细胞体外侵袭能力（P〈0．01），侵袭抑制率随脂联素重组体质量浓度的升高而增加，介于22．64％～77．84％之间；脂联素重组体质量浓度高于2．5μg／ml时，明显地抑制肿瘤细胞的迁移能力（P〈0．01）；脂联素重组体对ECM和FN黏附能力影响的程度不同，对FN的敏感性大于ECM；脂联素重组体质量浓度大于2．5μg／ml时，明显抑制肿瘤细胞MMP-2和MMP-9的分泌（P〈0、05或P〈0．01），后者的分泌量随质量浓度的升高而下降。结论：脂联素重组体在大于2．5μg／ml时，可能通过保护基底膜不受破坏而抑制人乳腺癌细胞的侵袭能力。     

Inhibition of invasion and metastasis by glypican-3 in a syngeneic breast cancer model

Glypican-3 (GPC3), a proteoglycan bound to the cell membrane through a GPI anchor, is widely expressed in the embryo but down regulated in most adult tissues, with some exceptions as mammary cells. GPC3 is involved in the regulation of cell proliferation and survival in specific cell types. LM3, a murine mammary tumor cell line unable to express GPC3, was stably transfected with the rat GPC3 gene to analyze its role in tumor progression. Upon injection into syngeneic BALB/c mice LM3-GPC3 clones showed less local invasiveness and developed fewer spontaneous and experimental lung metastasis than controls. GPC3-expressing cells were more sensitive to apoptosis induced by serum depletion, exhibited a delay in the first steps of spreading and were less motile than controls. On the other hand, LM3-GPC3 cells were significantly more adherent to FN than control ones. We observed that GPC3 transfectants presented a higher expression of E-cadherin and -catenin, molecules whose down regulation has been associated with tumor progression. Exogenous TGF- increased MMP-9 activity in both control and GPC3-expressing cells, but did not modulate MMP-2. Contrarily, GPC3 expression prevented the increase of MMP-2 activity induced by IGF-II. Our results suggest that GPC3 has a protective role against mammary cancer progression.     

Integrin-dependent response to laminin-511 regulates breast tumor cell invasion and metastasis

The basement membrane protein, laminin (LM)-511, is a potent adhesive and migratory substrate for metastatic breast tumor cells in vitro. Its expression correlates with tumor grade and metastatic potential in vivo. These observations suggest that responsiveness to autocrine or paracrine-derived LM-511 may be an important property regulating breast cancer metastasis in vivo. To address this, we compared the metastatic potential of 4T1 mammary carcinoma cells to that of 4T1 variants isolated by repeated chemotactic migration toward LM-511 in vitro (4T1LMF4) followed by serial injection into the mammary gland and recovery of spontaneous metastases from bone (4T1BM2). Variant subpopulations exhibited a distinct morphology on LM-511 and increased expression of β1 and β4 integrins compared to parental 4T1 cells. Importantly, mice inoculated with 4T1LMF4 and 4T1BM2 variants showed a 2.5- to 4-fold increase in the incidence of spontaneous metastasis to bone compared to 4T1 tumor-bearing mice. Functionally, 4T1BM2 variants were more adherent and more invasive toward LM-511 than parental 4T1 cells. Treatment of 4T1BM2 cells with lebein-1, a disintegrin with selectivity toward LM-type integrin receptors, potently inhibited their migration and invasion toward LM-511. Similarly, α3β1 integrin-dependent migration and invasion of human MDA-MB-231 breast carcinoma cells toward LM-511 were significantly inhibited by lebein-1. Taken together, these results provide strong evidence that LM-511 contributes to the metastasis of breast tumors and suggest that targeting integrin-LM-511 interactions with lebein-1 or other inhibitors of LM-511 receptors may have therapeutic potential for patients with advanced breast cancer.     

S100A4 regulates cell motility and invasion in an in vitro model for breast cancer metastasis

Elevated levels of the calcium-binding protein S100A4 are associated with poor patient survival in breast cancer patients and induce metastasis in rodent models. To investigate the effects of S100A4 on different components of the metastatic process, epithelial cells lines have been isolated from nonmalignant tumours in neu transgenic mice and from malignant tumours in neu/S100A4 double transgenic mice. Additional cell lines expressing both Neu and S100A4 have also been derived by transfection of rat S100A4 cDNA into tumour cell lines cloned from neu single transgenic mice. Using these cells in transfilter migration assays, it has been shown that increases in either motility or invasive properties correlate with each other and with the level of S100A4 protein. Injection of three of the cell lines separately into the mammary fat pads of nude mice showed that elevated levels of S100A4 correlated with the degree of metastasis to the lungs. In contrast, changes in cell proliferation and cell-substrate adhesion did not correlate with S100A4 levels. Neither motility nor invasiveness correlated with proteolytic degradation of gelatin as measured by zymography. Thus, the results suggest that the main effect of increases in S100A4 levels in metastasis is to generate increased cell motility and invasion and that this latter change is not dependent upon an increased ability to degrade the intercellular matrix.     

High-density lipoprotein of patients with type 2 diabetes mellitus elevates the capability of promoting migration and invasion of breast cancer cells

Epidemiological studies suggested complicated associations between type 2 diabetes mellitus and breast cancer. There is a significant inverse association between high-density lipoprotein (HDL) and the risk and mortality of breast cancer. However, HDL could be modified in various ways in diabetes patients, and this may lead to the altered effects on many different types of cells. In our study, we found that glycation and oxidation levels are significantly higher in HDL from type 2 diabetes mellitus patients compared to that from healthy subjects. Diabetic HDL dramatically had a stronger capability to promote cell proliferation, migration and invasion of breast cancer (as examined both on hormone-independent cells and on hormone-dependent cells). In addition, glycated and oxidized HDL, which were produced in vitro, acted in similar way as diabetic HDL. Diabetic HDL, glycated HDL and oxidized HDL also induced higher synthesis and secretion of VEGF-C, MMP-2 and MMP-9 from malondialdehyde (MDA)-MB-231 cells. It was indicated that diabetic, glycated and oxidized HDL promote MDA-MB-231 cell migration and invasion through ERK and p38 MAPK pathways, and Akt pathway plays an important role as well in MDA-MB-231 cell invasion. The Akt, ERK and p38 MAPK pathways are also involved in VEGF-C and MMP-9 secretion induced by diabetic, glycated and oxidized HDL. Our study demonstrated that glycation and oxidation of HDL in diabetic patients could lead to abnormal actions on MDA-MB-231 cell proliferation, migration and invasion, thereby promoting the progression of breast cancer. This will largely draw the attention of HDL-based treatments in diabetic patients especially those with breast cancer.     

基质金属蛋白酶及其抑制剂与肺癌的侵袭和转移

基质金属蛋白酶(MMPs)家族是细胞外基质降解过程中的重要酶类,与多种病理过程尤其是肿瘤侵袭和转移有密切关系,因此成为肿瘤治疗的新靶点,本文就MMPs、TIMPs的生物学作用,与肺癌侵袭转移的关系及MMP合成抑制剂的开发研究情况等方面做一综述.     

MMPs与乳腺癌的研究进展

目的:研究基质金属蛋白酶(MMPs)在乳腺癌发病进展过程中所起的作用及其机制。方法:应用PubMed、Springer期刊检索系统,以"乳腺癌、基质金属蛋白酶、细胞外基质、侵袭和转移"等为关键词,检索1997-2009年的相关文献,共检索到英文文献100条。纳入标准:MMPs的分类,结构及一般特性;MMPs与乳腺癌发生发展的关系;MMPs在乳腺癌临床诊断及治疗中的应用。符合分析的文献39篇。结果:在肿瘤发生的早期,MMPs通过促进血管生成、活化生长因子来促进肿瘤的发生进展,而在后期通过降解细胞外基质来促进肿瘤的侵袭和转移。MMPs因其作用的复杂性,目前研制的MMPs抑制剂均效果不佳。结论:MMPs及其抑制物在肿瘤发生发展各期所起的作用不尽相同,研究其各自的作用,开发特异性的抑制物将为乳腺癌临床治疗提供新方向。     

Triptolide inhibits matrix metalloproteinase-9 expression and invasion of breast cancer cells through the inhibition of NF-κB and AP-1 signaling pathways

Triptolide is a diterpenoid epoxide that is endogenously produced by the thunder god vine, Tripterygium wilfordii Hook F. Triptolide has demonstrated a variety of biological activities, including anticancer activities, in previous studies. Invasion and metastasis are the leading causes of mortality for patients with breast cancer, and the increased expression of matrix metalloproteinase-9 (MMP-9) has been shown to be associated with breast cancer invasion. Therefore, the aim of the present study was to investigate the effect of triptolide on 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced cell invasion and MMP-9 expression in breast cancer cells. The expression of signal molecules was examined by western blotting, zymography and quantitative polymerase chain reaction; an electrophoretic mobility gel shift assay was also used, and cell invasiveness was measured by an in vitro Matrigel invasion assay. The MCF-7 human breast cancer cell line was treated with triptolide at the highest concentrations at which no marked cytotoxicity was evident. The results demonstrated that triptolide decreased the expression of MMP-9 through inhibition of the TPA-induced phosphorylation of extracellular signal-regulated kinase (ERK) and the downregulation of nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) activity. In addition, a Transwell assay revealed that triptolide reduced the ability of MCF-7 cells to invade Matrigel. These data demonstrate that the anti-invasive effect of triptolide is associated with the inhibition of ERK signaling and NF-κB and AP-1 activation, and suggest that triptolide may be a promising drug for breast cancer.