快速老化小鼠海马中细胞外基质金属蛋白酶诱导因子的年龄相关性变化

目的:观察细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,CD147)在快速老化小鼠(senescence-accelerated-prone mouse 8,SAMP8)海马中随年龄变化的趋势。方法:应用免疫组织化学和Western blot的方法,观察CD147在SAMP8小鼠海马中的年龄相关性变化。结果:免疫组织化学结果显示SAMP8小鼠海马区CD147免疫反应呈阳性,且随年龄增加无明显的细胞分布变化。Western blot分析数据表明随年龄增加CD147蛋白水平逐渐下降;相对于3月龄小鼠,12月龄小鼠的CD147蛋白水平下降了约53%(P<0.05)。结论:随年龄的增加,海马区CD147蛋白水平逐渐下降,推测其与阿尔茨海默病(Alzheimer’s disease,AD)相关。     

Extracellular matrix metalloproteinase inducer in interstitial pneumonias

Extracellular matrix metalloproteinase inducer (EMMPRIN), a glycosylated transmembrane protein that induces matrix metalloproteinases (MMPs), is minimally expressed in the normal adult lung. We previously reported that it is up-regulated in murine bleomycin-induced lung injury. In this study, we determined the expression of EMMPRIN and its association with MMP-2, MMP-7, and MMP-9 in interstitial pneumonias (IPs). We performed immunohistochemistry for EMMPRIN and MMPs on lung tissue from 22 subjects with various IPs. We did bronchoalveolar lavage (BAL) on 9 of these subjects and 13 others with IPs to measure the soluble EMMPRIN in BAL fluid. For comparison, immunohistochemistry or BAL was done on 14 subjects without IPs. The staining intensity for each protein was scored from 0 to 3 in various epithelial cell types. Soluble EMMPRIN in BAL fluid was measured by an enzyme-linked immunosorbent assay. Extracellular matrix metalloproteinase inducer was prominent in abnormal epithelial cells. It was more prominent in hyperplastic type II cells, compared with epithelium in alveolar bronchiolization. It was also elevated in BAL fluid from the subjects with IPs. Matrix metalloproteinases were expressed in cells expressing EMMPRIN, although the profile of MMPs varied among the different abnormal epithelial cell types with MMP-2 and MMP-7 in hyperplastic type II cells and MMP-7 and MMP-9 in cells showing squamous metaplasia and cells comprising bronchiolization. These results suggest a role of EMMPRIN in reepithelialization in IPs.     

PPARs配体对巨噬细胞、泡沫细胞细胞外基质金属蛋白酶诱导因子表达的影响

目的：观察PPAR α、γ配体对巨噬细胞、泡沫细胞细胞外基质金属蛋白酶诱导因子(EMMPRIN)表达的影响。方法：体外诱导THP-1单核细胞转化为巨噬细胞、泡沫细胞，分别加入PPAR α配体氯贝特（clofibrate）、PPARγ配体吡格列酮(pioglitazone)共同培养，应用Real-time RT-PCR和Western blotting测定巨噬细胞、泡沫细胞中EMMPRIN基因和蛋白表达，ELISA测定细胞培养上清液MMP-9浓度，Zymgraphy法测定MMP-9活性。结果：氯贝特和吡格列酮均能显著抑制巨噬细胞和泡沫细胞EMMPRIN的表达，此抑制作用与PPAR α、γ配体抑制MMP-9分泌及活性的趋势一致。结论：PPAR α、γ配体均可抑制巨噬细胞、泡沫细胞EMMPRIN的表达，下调EMMPRIN可能是PPARs配体抑制粥样斑块局部MMPs产生的机制之一。     

成纤维细胞与结肠癌细胞相互作用促进细胞外基质金属蛋白酶诱导因子的表达

目的研究成纤维细胞与结肠癌细胞相互作用对二者表达细胞外基质金属蛋白酶诱导因子（EMMPRIN）的影响,初步探讨肿瘤-基质相互作用在结肠癌侵袭转移中的作用。方法结肠癌SW480细胞与HELF成纤维细胞以RPMI1640培养液分别共培养0、12、24、48h,通过RT-PCR和免疫细胞化学方法检测SW480和HELF细胞中EMMPRIN的表达。结果共培养组SW480细胞EMMPRINmRNA和蛋白的表达均明显升高,HELF细胞本来不表达EMMPRIN,与SW480细胞共培养12、24、48h后检测到EMMPRIN表达,且随时间延长而表达增加。结论成纤维细胞与结肠癌细胞相互作用上调SW480细胞EMMPRIN的表达,并诱导HELF细胞表达EMMPRIN,有可能在结肠癌侵袭转移中发挥重要作用。     

Progesterone receptor modulator CDB-2914 induces extracellular matrix metalloproteinase inducer in cultured human uterine leiomyoma cells

Effects of progesterone receptor modulator CDB-2914 on the expression of the extracellular matrix (ECM) components were examined in cultured human uterine leiomyoma and myometrial cells. ECM metalloproteinase inducer (EMMPRIN), matrix metalloproteinases (MMPs), tissue inhibitors of MMP (TIMPs) and collagen levels were assessed by Western blot analysis, MMP activity assay and real-time RT-PCR. RNA interference (RNAi) of EMMPRIN was performed using small interfering mRNA. In cultured leiomyoma cells, CDB-2914 treatment at concentrations greater than or equal to 10(-8) M significantly increased EMMPRIN, MMP-1 and MMP-8 protein contents and MMP-1, MMP-2, MMP-3 and MMP-9 mRNA levels, and activity of MMP-1, MMP-2, MMP-3 and MMP-9 in the medium. TIMP-1 and TIMP-2 were significantly decreased at mRNA and protein levels by CDB-2914 treatment at concentrations > or =10(-7) M in these cells. CDB-2914 treatment decreased types I and III collagen protein contents. However, CDB-2914 treatment did not affect the ECM component expression in cultured myometrial cells. RNAi of EMMPRIN abrogated CDB-2914-mediated both induction of MMPs and reduction of TIMPs and collagens in cultured leiomyoma cells. These results suggest that CDB-2914 modulates the expression of EMMPRIN, MMPs, TIMPs and collagens in cultured leiomyoma cells without comparable effects on myometrial cells.     

Differential expression of extracellular matrix metalloproteinase inducer (CD147) in normal and ulcerated corneas: role in epithelio-stromal interactions and matrix metalloproteinase induction

Extracellular matrix metalloproteinase inducer (EMMPRIN) was originally identified on the tumor cell surface as an inducer of matrix metalloproteinase (MMP) production in neighboring fibroblasts. Here we demonstrate a role for EMMPRIN in MMP induction during corneal wound healing. MMP and EMMPRIN expression was analyzed in normal and ulcerated human corneas, as well as in corneal epithelial and stromal cells in culture using confocal microscopy, zymography, immunoblots, and real-time polymerase chain reaction. In normal cornea EMMPRIN was predominantly expressed in the epithelium but was markedly induced in the anterior stroma of ulcerated corneas. This coincided with MMP-2 induction that co-localized with EMMPRIN at the epithelio-stromal boundary. The role of epithelial-stromal interaction in MMP induction was investigated in an in vitro co-culture system and demonstrated an induction and co-localization of EMMPRIN and MMP-2 in the fibroblasts at the interface with epithelial cells. Direct contact of fibroblasts with EMMPRIN-containing purified epithelial cell membranes also induced MMP-1, MMP-2, and EMMPRIN and this was inhibited by a blocking anti-EMMPRIN antibody, suggesting that EMMPRIN was primarily responsible for this induction. These findings, and the up-regulation of EMMPRIN by epidermal growth factor and transforming growth factor-beta, demonstrate a role for EMMPRIN in wound healing and suggest that sustained local up-regulation of EMMPRIN and MMPs in chronic situations in which healing is delayed may lead to excessive matrix degradation and corneal melts.     

Differential expression of extracellular matrix metalloproteinase inducer (EMMPRIN/CD147) in avian tibial dyschondroplasia

Tibial dyschondroplasia (TD) is an avian bone disorder of different aetiologies that may be associated with lameness. The disorder is characterized by focal disruption of endochondral bone formation, with a lack of matrix proteolysis and an accumulation of non-mineralized avascular cartilage. The aim of this study was to determine the expression of extracellular matrix metalloproteinase inducer (EMMPRIN/CD147) in normal, thiram-induced TD lesions and in the process of recovery from TD in broiler chickens. An extracellular matrix (ECM) degrading enzyme, matrix metalloproteinase-9 (MMP-9), was selected to investigate the effects of CD147 in the degradation of ECM. Gene expression was analysed by quantitative real-time polymerase chain reaction and protein levels by immunohistochemistry and western blotting. The birds were divided into three groups: thiram fed; recovery; and controls. Genes encoding CD147 and MMP-9 were down-regulated during the development of the disease, and were up-regulated during recovery. Western blotting also showed lower protein levels of CD147 in TD, which increased during the recovery phase associated with ECM degradation and growth plate repair. The findings of this study suggest that ECM has a crucial role in the occurrence of TD and that CD147 appears to play a pivotal role in matrix proteolysis in the chicken, similar to that in other species.     

人参皂甙Rh_2对耐药乳腺癌细胞P-糖蛋白和基质金属蛋白酶及其诱导物表达的影响

目的观察人参皂甙Rh2对人乳腺癌MCF7/Adr细胞P-糖蛋白(P-glycoprotein,P-gp)、基质金属蛋白酶诱导物(extracel-lular matrix metalloproteinase inducer,EMMPRIN)和基质金属蛋白酶1(matrix metalloproteinases1,MMP1)、MMP2、MMP9表达的影响。方法采用MTT比色法检测药物对MCF7/Adr细胞的毒性作用;流式细胞仪测定细胞内阿霉素(adriamycin,ADM)的荧光强度;Real time RT-PCR和Western blot检测细胞P-gp、EMMPRIN、MMP1、MMP2和MMP9mRNA和蛋白水平的变化。结果 ADM组与对照组相比,ADM能上调细胞P-gp、EMMPRIN、MMP1、MMP2和MMP9蛋白表达;人参皂甙Rh2能抑制ADM对细胞P-gp、EMMPRIN、MMP1、MMP2和MMP9蛋白和mRNA水平的上调作用。结论人参皂甙Rh2可以有效逆转乳腺癌细胞耐药细胞系MCF7/ADM的耐药性。     

EMMPRIN (extracellular matrix metalloproteinase inducer) is a novel marker of poor outcome in serous ovarian carcinoma

EMMPRIN is a member of the immunoglobulin superfamily of adhesion molecules and has a role in the activation of several matrix metalloproteinases (MMP). The objective of this study was to investigate the expression of EMMPRIN in effusions, primary and metastatic tumors of serous ovarian carcinoma patients, as well as to evaluate its association with clinicopathologic parameters and with MMP and integrin expression. Eighty effusions and eighty-three solid lesions were evaluated for expression of EMMPRIN mRNA using in situ hybridization (ISH). Protein expression was studied in 75 effusions and 55 biopsies using immunohistochemistry (IHC). EMMPRIN mRNA and protein were detected in carcinoma cells in 63/80 (79%) and 64/75 (85%) effusions, respectively. Expression was similar in peritoneal and pleural effusions. EMMPRIN was co-expressed with MMP-1 (P<0.001), MMP-9 (P=0.006) and the αv (P=0.013) and β1 (P=0.029) integrin subunits. In solid lesions, EMMPRIN localized most often to tumor cells (51/83 using ISH, 51/55 using IHC), but was also expressed in stromal and endothelial cells in approximately one third of the cases. EMMPRIN mRNA expression in tumor cells was most frequent in peritoneal metastases (P=0.03). EMMPRIN expression in carcinoma cells of solid tumors showed an association with that of MMP-9 (P=0.018), while labeling of stromal cells showed co-localization with the β1 integrin subunit (P=0.043). In survival analysis, EMMPRIN protein expression in stromal cells of primary tumors (P=0.012) and in endothelial cells of all solid tumors (P=0.023) correlated with poor survival. In conclusion, EMMPRIN is a novel prognostic marker in ovarian carcinoma, and is co-expressed with other metastasis-associated molecules in this malignancy. The identical phenotype of carcinoma cells in pleural and peritoneal effusions provides further evidence to our theory that cells at these sites share similar genotypic and phenotypic profiles. This revised version was published online in July 2006 with corrections to the Cover Date.     

高糖和茶多酚对人脐静脉内皮细胞基质金属蛋白酶2及其诱导因子表达的影响

目的 探讨高糖对内皮细胞基质金属蛋白酶2(MMP-2)及其诱导因子(EMMPRIN)表达的影响,以及茶多酚对高糖作用的干预情况.方法 培养人脐静脉内皮细胞,分为正常对照组、高糖组、茶多酚对照组和茶多酚干预组.培养24 h后,用免疫细胞化学、RT-PCR和Western blot法测定细胞内MMP-2及EMMPRIN的变化.结果 高糖下调内皮细胞MMP-2及EMMPRIN表达,其变化随时间延长更为明显;而茶多酚干预组MMP-2活性明显上调(P<0.05);茶多酚干预组EMMPRINmRNA及蛋白表达均较高糖组上调(P<0.05),茶多酚干预可拮抗高糖对内皮细胞的影响.结论 高糖影响内皮细胞的细胞外基质降解,茶多酚可以抑制这种作用.     

Cytokines regulate matrix metalloproteinases in human uterine endometrial fibroblast cells through a mechanism that does not involve increases in extracellular matrix metalloproteinase inducer

PROBLEM: Endometriosis is the presence of ectopic uterine endometrial tissue in the peritoneal cavity. Peritoneal fluid samples of women with endometriosis show elevated interleukin-1 (IL-1)beta, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta(1) levels, indicating that an altered immune system may play an important role in the pathogenesis of endometriosis. The invasion of ectopic endometrium into peritoneal mesothelium requires matrix metalloproteinases (MMPs) for tissue remodeling. Several MMPs are differentially expressed in human uterine endometrium with menstrual endometrium showing the highest level of expression. MMPs are stimulated by cytokines and also by the protein Extracellular Matrix Metalloproteinase Inducer (EMMPRIN). METHOD OF STUDY: To determine the role of cytokines in ectopic endometrial invasion, we investigated whether cytokines could regulate MMP production by endometrial fibroblast cells and whether this stimulation occurred through an effect on EMMPRIN expression. Human uterine fibroblasts (HUF) were treated with IL-1beta, TGF-beta(1) and TNF-alpha in a dose dependent and time dependent manner (C, 0.1, 1, 10 ng/mL IL-1beta or TGF-beta(1); C, 2, 10, 50 ng/mL TNF-alpha) for 0, 6, 12, and 24 hr. Cell conditioned medium samples were collected and concentrated at each timepoint for immunoblot analysis. Cellular RNA was collected for real time PCR analysis of MMPs-1, -2, -3 and EMMPRIN mRNA levels. RESULTS: Our results showed that IL-1beta stimulated MMP-1 protein secretion and mRNA levels in a time dependent manner (P < 0.05), MMP-2 mRNA in a time dependent manner and MMP-3 in a time and dose dependent manner. TNF-alpha stimulated MMP-1 and -3 protein secretion in a time dependent manner and stimulated MMP-1, -2 and -3 mRNA levels in a time dependent manner (P < 0.05). Neither IL-1beta nor TNF-alpha treatment affected MMP-2 protein secretion. TGF-beta(1) inhibited MMP-1 and MMP-2 mRNAs at the highest treatment dose after 24 hr but there was no effect on protein secretion. TGF-beta(1) exerted no effect on MMP-3 mRNA or protein secretion (P < 0.05). Neither of the cytokines affected EMMPRIN protein or mRNA levels but the 10 ng/mL TGF-beta(1) treatment did cause a reduction in EMMPRIN mRNA levels. CONCLUSIONS: These data show that elevated cytokines may play a role in the establishment of ectopic endometrium in the peritoneal cavity by stimulating MMPs to remodel the mesothelial lining of the peritoneum thus allowing for tissue invasion. The stimulation of MMPs by cytokines occurred without any change in EMMPRIN expression whereas the inhibitory effect of TGF-beta(1) involved a reduction in EMMPRIN mRNA levels.     

High extracellular matrix metalloproteinase inducer/CD147 expression is strongly and independently associated with poor prognosis in colorectal cancer

As in most solid tumors, colorectal cancer prognosis strongly depends on the extent of local invasion and lymph node and distant metastases. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a transmembrane glycoprotein that activates matrix metalloproteinases, a group of enzymes that play an important role in tumor invasion and metastasis formation. This study investigates the EMMPRIN expression in a large cohort of patients with colorectal cancer. Immunohistochemical analysis of tissue microarrays from 285 patients shows that increased EMMPRIN protein expression does not correlate with clinicopathologic parameters and is an independent prognostic factor of poor survival, with mean survival times of 103 months in EMMPRIN negative/low versus 57 months in EMMPRIN intermediate/high patients (P < .001). This pronounced association of increased EMMPRIN levels and--on average--a 45% reduction in overall survival could help improve the risk stratification in patients with colorectal cancer; moreover, the lack of correlations with classical measures of cancer invasion/spreading may suggest the relevance of alternative EMMPRIN pathways beyond matrix metalloproteinase activation.     

Selective progesterone receptor modulator asoprisnil down-regulates collagen synthesis in cultured human uterine leiomyoma cells through up-regulating extracellular matrix metalloproteinase inducer

BACKGROUND: A recent clinical trial demonstrated that selective progesterone receptor modulator asoprisnil is effective in reducing uterine leiomyoma volume. We investigated the effects of asoprisnil in vitro on the expression of the extracellular matrix (ECM)-remodeling enzymes and collagens in cultured leiomyoma and matching normal myometrial cells. METHODS: The expression of extracellular matrix metalloproteinase inducer (EMMPRIN), matrix metalloproteinases (MMPs), tissue inhibitors of MMP (TIMPs) and collagens were assessed by western blot analysis. RESULTS: Untreated cultured leiomyoma cells had significantly lower EMMPRIN (P < 0.05), MMP-1 (P < 0.05) and membrane type 1-MMP (MT1-MMP) (P < 0.01) protein contents, but significantly higher TIMP-1 (P < 0.05), TIMP-2 (P < 0.01), type I (P < 0.05) and type III (P < 0.01) collagen protein contents compared with untreated cultured myometrial cells. Treatment with asoprisnil at concentrations > or =10(-7) M for 48 h significantly (P < 0.05) increased EMMPRIN, MMP-1 and MT1-MMP protein contents, and decreased TIMP-1 (P < 0.05), TIMP-2 (P < 0.01), type I (P < 0.01) and type III (P < 0.05 at 10(-7) M; P < 0.01 at 10(-6) M) collagen protein contents in cultured leiomyoma cells compared with control cultures. However, asoprisnil treatment did not affect the protein contents of ECM-remodeling enzymes and collagens in cultured myometrial cells. CONCLUSIONS: These results suggest that asoprisnil may reduce collagen deposit in the ECM of cultured leiomyoma cells through decreasing collagen synthesis and enhancing the expression of EMMPRIN, MMPs and TIMPs without comparable effects on cultured myometrial cells.     

细胞外基质金属蛋白酶诱导因子和内皮素转化酶对非小细胞肺癌患者术后淋巴结转移和预后的影响

目的： 研究细胞外基质金属蛋白酶诱导因子(EMMPRIN)和内皮素转化酶(ECE)在非小细胞肺癌（NSCLC）组织中表达的临床意义。 方法： 用免疫组织化学方法检测77例NSCLC组织中EMMPRIN和ECE的表达情况，分析其与肿瘤大小、癌的组织学类型、组织分化程度、淋巴结转移和预后的关系。 结果： 77例NSCLC组织中EMMPRIN和ECE的阳性表达率分别为66%和45%。EMMPRIN和ECE的表达均与淋巴结转移呈正相关（均P＜0.01，r=0.371和0.467），两者表达阳性组术后生存期均明显短于阴性组（均P＜0.01）。NSCLC组织中EMMPRIN和ECE的表达之间呈正相关（P＜0.05，r=0.243）。EMMPRIN和ECE的表达与肿瘤大小、癌组织学类型和组织分化程度均无相关性（P>0.05）。 结论： EMMPRIN和ECE的表达与NSCLC的淋巴结转移和预后密切相关，它们的高表达提示非小细胞肺癌患者预后不良。     

非小细胞肺癌中EMMPRIN和HGF的表达及其对淋巴结转移和预后的影响

目的： 研究细胞外基质蛋白酶诱导因子(EMMPRIN)和肝细胞生长因子(HGF)在非小细胞肺癌（NSCLC）组织中的表达及其临床意义。方法： 免疫组织化学方法检测77例NSCLC组织中EMMPRIN与HGF的表达，分析其与患者吸烟情况、肿瘤大小、组织学类型、组织分化程度、淋巴结转移和预后的关系。 结果：77例NSCLC组织中EMMPRIN与HGF的阳性表达率分别为68%和44%。EMMPRIN与HGF的表达均与淋巴结转移呈正相关（r=0.371和0.339，P＜0.01），与患者术后生存时间呈负相关（P＜0.01）。EMMPRIN与HGF的表达均与患者吸烟、肿瘤大小、组织学类型和组织分化程度无关（P＞0.05）。EMMPRIN与HGF的表达之间呈正相关（r=0.281,P＜0.01）。结论: EMMPRIN和HGF的表达与NSCLC的淋巴结转移和预后密切相关，它们的高表达提示NSCLC患者预后不良。     

Expression of gelatinase B and the extracellular matrix metalloproteinase inducer EMMPRIN in benign and malignant pigment cell lesions of the skin.

By the degradative effect on basement membrane collagen type IV, matrix metalloproteinases (MMPs) or gelatinases are important in the early invasion of malignant tumors. These enzymes may be released by the tumor cells themselves or may be derived from nearby fibroblasts that have been stimulated by the extracellular MMP inducer EMMPRIN. We studied the distribution of 92-kd gelatinase B (MMP-9) and of EMMPRIN in 33 benign and 41 malignant, paraffin-embedded pigment cell lesions using immunohistochemistry and monoclonal antibodies. In benign pigment cell lesions, EMMPRIN but not gelatinase B was expressed in cellular blue nevi whereas all other benign lesions, including common blue nevi, were negative. In malignant melanomas (MMs), both gelatinase B and EMMPRIN were variably expressed in the pure and invasive radial growth phase but not in the vertical growth phase. All lentigo maligna cases and all metastatic lesions were negative. Of MMs with thickness < 1.6 mm, 63% expressed gelatinase B and 70% expressed EMMPRIN, whereas in MMs with > 1.6 mm thickness, only 10% expressed gelatinase B and only 25% expressed EMMPRIN. We conclude that early invasion of MM is associated with de novo expression of gelatinase B and EMMPRIN by neoplastic melanocytes. Expression of EMMPRIN and MMP-9 may be partly responsible for the stromal changes observed in thin MM. Their absence in the vertical growth phase and in metastatic lesions suggests that other factors are involved in tissue degradation during later stages of tumor progression in MM. The lack of both gelatinase B and EMMPRIN in lentigo maligna may contribute to the indolent behavior of this type of pigment cell lesion.     

细胞外基质金属蛋白酶诱导因子降解途径的初步研究

目的:观察蛋白酶体抑制剂MG-132对细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,CD147)表达的影响,探讨CD147的降解途径。方法:在SY5Y细胞中分别加入0,1,2.5和5μmol/L不同浓度剂量的蛋白酶体抑制剂MG-132作用24 h后,通过Western Blot和In-cell Western的方法检测CD147的相对含量;免疫荧光双标检测SY5Y细胞中CD147与泛素(ubiquitin)的表达及共定位情况。结果:在一定时间内,随着MG-132浓度的增大,CD147的含量相对增多;与0μmol/L组比较,5μmol/L组的CD147含量在两种实验方法中分别增加了约151.81%±36.26%(P0.05)和76.43%±18.02%(P0.05);CD147与泛素在细胞胞体中存在共定位的特征。结论:蛋白酶体抑制剂MG-132可增加CD147的含量;CD147可被泛素化。提示CD147可能通过泛素-蛋白酶体途径进行降解。