EMMPRIN基因增强CT26小鼠结肠癌细胞的转移潜能

背景与目的:结肠癌的侵袭转移是临床上绝大多数结肠癌患者的致死因素,研究表明,细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,EMMPRIN)基因参与了肿瘤的转移过程.为研究EMMPRIN基因在结肠癌侵袭转移过程中的意义,本研究将初步探讨EMMPRIN基因对CT26小鼠结肠癌细胞转移潜能的影响.方法:构建包含EMMPRIN基因编码框的真核表达载体pCMY-HA2-EMMPRIN,通过质脂体转染CT26细胞,经G418筛选,建立稳定表达EMMPRIN基因的CT26细胞株.通过体外侵袭实验、迁移实验、粘附实验分析EMMPRIN在CT26中过表达及CT26细胞在侵袭、迁移、粘附等转移能力上的改变.结果:当EMMPRIN基因在CT26细胞稳定过表达后,侵袭实验的结果为EMMPRIN组:103.33±8.49个细胞,对照组:48.67±5.31个细胞(P<0.01);迁移实验的结果为EMMPRIN组:40.67±2.49个细胞,对照组:18.33±2.05个细胞(P<0.01);粘附实验的CCK-8检测结果为EMMPRIN组:2.10±0.22,对照组:3.33±0.17(P<0.01).结论:EMMPRIN基因可明显促进小鼠结肠癌细胞CT26的侵袭与迁移,并抑制其粘附,提示EMMPRIN基因对CT26细胞的转移具有促进作用.     

Increased EMMPRIN (CD 147) expression during oral carcinogenesis

Gene expression profiling of oral premalignant (OPM) cells and normal oral epithelial (NOR) cells showed that EMMPRIN expression was markedly upregulated in OPM cells compared to NOR cells. We used an oral squamous cell carcinoma (OSCC) progression model composed of cell lines, organotypic cultures and tissue specimens to characterize EMMPRIN expression patterns by microarray analysis, qRT-PCR, Western blotting and immunohistochemistry. EMMPRIN levels are elevated in OPM and primary and metastatic OSCC cells as compared to NOR. EMMPRIN was detected as high and low glycosylated forms in the OPM and OSCC cellular extracts and was released in the media by OSCC cells but not by OPM cells. EMMPRIN expression in an organotypic culture model of normal and OPM mucosae mirrored the expression patterns in the respective tissues in vivo. EMMPRIN expression was limited to basal cells of normal, benign hyperkeratotic and inflammatory (lichen planus) oral mucosa. EMMPRIN expression is increased in dysplastic leukoplakias spreading to more superficial layers, and its expression levels correlated significantly with the degree of dysplasia. Primary and metastatic OSCC showed strong cell surface expression of EMMPRIN. These results suggest that EMMPRIN overexpression occurs at a very early stage of oral carcinogenesis and plays a contributing role in OSCC tumorigenesis.     

A soluble factor (EMMPRIN) in exudate influences knee motion after total arthroplasty

Few studies have been conducted to investigate biological factors that affect postoperative knee motion after total knee arthroplasty (TKA). The purpose of this study is to test the hypothesis that range of knee motion (ROM) at 4 weeks after TKA is correlated with the concentration of extracellular matrix metalloproteinase inducer (EMMPRIN) and transforming growth factor (TGF)-beta1 in the exudative fluid harvested from the joint after surgery. A prospective measurement study was conducted with 20 osteoarthritis patients who underwent TKA. At 48 h after surgery, the exudate was harvested from a closed drainage system. Enzyme-linked immunosorbent assay was performed to measure the concentration of TGF-beta1, EMMPRIN, MMP-1, 2, 9, tissue inhibitor of metalloproteinase-1, and Hyalunonan. Knee flexion angle was measured before and at 4 weeks after surgery. There was a significant correlation between the EMMPRIN levels and knee flexion angle (r = 0.557, p = 0.0148). Western blot analysis of the exudate showed a prominent band for EMMPRIN at 27 kDa. On the other hand, there was no correlation between the TGF-beta1 levels and the knee flexion angle. This study showed that EMMPRIN levels after TKA affect the postoperative ROM. As to clinical relevance, EMMPRIN in the exudate after TKA is a promising biological indicator to predict difficulty in restoring postoperative ROM.     

CD147与类风湿关节炎成纤维样滑膜细胞MMPs合成的关系

目的观察类风湿性关节炎(rheumatoid arthritis, RA)患者成纤维样滑膜细胞(fibroblast-like synoviocytes, FLS)与高表达CD147的单核细胞株(THP-1)共培养后,基质金属蛋白酶(matrix metalloproteinases,MMPs)合成的变化.方法从手术切除的RA患者滑膜组织中,分离FLS,经流式细胞术测定CD14,CD68及免疫组化测定波形纤维蛋白(vimentin)证实后,传代培养.将FLS与高表达 CD147的THP-1共培养,观察THP-1细胞对FLS MMPs合成的影响.用明胶酶谱法测FLS MMP-2,MMP-9含量.同时观察CD147拮抗肽(AP9)对MMPs表达的影响.结果高表达CD147,低表达MMP-2、 MMP-9的THP-1细胞与CD147表达水平低的RA FLS混合培养后,FLS合成MMP-2、 MMP-9水平显著升高,随着THP-1细胞数的增加,MMP-2、 MMP-9的合成量也增加.THP-1对FLS合成MMP-2、 MMP-9的刺激作用可被CD147拮抗肽AP-9所抑制.结论高表达CD147的THP-1细胞对共培养的RA FLS合成MMP-2、MMP-9有刺激作用,这种刺激作用可被CD147拮抗肽AP-9所抑制,提示THP-1表面的CD147对FLS合成MMPs有促进作用.     

MMP-14与EMMPRIN在口腔鳞状细胞癌中的表达及临床意义

目的：探讨基质金属蛋白酶-14(matrix metalloproteinase 14,MMP-14)和细胞外基质金属蛋白酶诱导因子（extracellular matrix metalloproteinase inducer,EMMPRIN）在口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）中的临床意义,并分析二者的相关性。方法选取2012年1月—2014年6月于我院口腔颌面外科诊治的69例OSCC患者作为OSCC组,同时选取43例口腔正常新鲜组织标本作为对照组。采用逆转录-聚合酶链反应技术检测组织中的MMP-14及EMMPRIN mRNA的表达含量。结果MMP-14 mRNA、EMMPRIN mRNA在OSCC组中的相对表达量明显高于对照组,差异具有统计学意义（t=8.198、9.624,P＜0.01）;OSCC组患者在年龄、性别因素中的MMP-14、EMMPRIN mRNA的表达差异不明显（P＞0.05）,在伴有淋巴结转移的患者中均明显升高（t=2.909、2.156,P＜0.05）;且MMP-14、EMMPRIN mRNA在Ⅲ～Ⅳ期、中低分化的OSCC中表达均明显升高,差异具有统计学意义（t=3.376、2.667、3.027、2.399,P＜0.05）;MMP-14与EMMPRIN mRNA表达含量间相关系数为r=0.821,二者呈正相关性（P＜0.01）。结论OSCC组织中MMP-14和EMMPRIN mRNA的高表达状态均与肿瘤的病理分期、分化程度及淋巴结转移因素关系密切,二者的表达具有正相关性,提示MMP-14和EMMPRIN均参与了OSCC的浸润及转移过程,通过检测EMMPRIN及MMP-14 mRNA表达含量可能有助于OSCC的病变程度及预后转归判断。     

Hepatocytes in liver injury: Victim,bystander, or accomplice in progressive fibrosis?

Chronic liver disease causes significant morbidity and mortality through progressive fibrosis, cirrhosis, and liver cancer. The classical theory of fibrogenesis has hepatic stellate cells (HSCs) as the principal and only significant source of abnormal extracellular matrix (ECM). Further, HSCs have the major role in abnormal ECM turnover. It is the death of hepatocytes, as the initial target of injury, that initiates a sequence of events including the recruitment of inflammatory cells and activation of HSCs. Following this initial response, the ongoing insult to hepatocytes is regarded as perpetuating injury, but otherwise, hepatocytes are regarded as “victims” and “bystanders” in progressive fibrosis. Recent developments, however, challenge this view and suggest the concept of the hepatocyte being an active participant in liver injury. It is clear now that hepatocytes undergo phenotypic changes, adapt to injury, and react to the altered microenvironment. In this review, we describe studies showing that hepatocytes contribute to progressive fibrosis by direct manipulation of the surrounding ECM and through signaling to effector cells, particularly HSCs and intrahepatic immune cells. Together, these findings suggest an active “accomplice” role for the hepatocyte in progressive liver fibrosis and highlight novel pathways that could be targeted for development of future anti‐fibrotic therapies.     

A novel role of EMMPRIN/CD147 in transformation of quiescent fibroblasts to cancer-associated fibroblasts by breast cancer cells

We tested the novel hypothesis that EMMPRIN/CD147, a transmembrane glycoprotein overexpressed in breast cancer cells, has a previously unknown role in transforming fibroblasts to cancer-associated fibroblasts, and that cancer-associated fibroblasts in turn induce epithelial-to-mesenchymal transition of breast cancer cells. Co-culture of fibroblasts with breast cancer cells or treatment of fibroblasts with breast cancer cell conditioned culture medium or recombinant EMMPRIN/CD147 induced expression of α-SMA in the fibroblasts in an EMMPRIN/CD147-dependent manner and promoted epithelial-to-mesenchymal transition of breast cancer cells and enhanced cell migration potential. These findings support a novel role of EMMPRIN/CD147 in regulating the interaction between cancer and stroma.     

Iron deficiency activates pro-inflammatory signaling in macrophages and foam cells via the p38 MAPK-NF-κB pathway

Background/objectives

Major bleeding in patients with acute coronary syndrome (ACS) increases the risk of recurrent ACS and mortality. However, the mechanism involved is poorly understood. Bleeding induces iron deficiency. Iron deficiency enhances inflammation in other diseases. Thus, in this paper, the particular effect of iron deficiency on atherosclerotic plaque destabilization, especially the pro-inflammatory role of iron deficiency in atheroma and the mechanism involved were investigated.

Methods

Extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase-9 (MMP-9) mRNA levels were investigated by RT-PCR. EMMPRIN and MMP-9 protein levels, nuclear factor (NF)-κB-p65 protein levels, peroxisome proliferator-activated receptor γ (PPARγ) protein levels, and mitogen-activated protein kinase (MAPK) phosphorylation were determined by western blotting. MMP-9 enzymatic activity was assayed by gelatin zymography.

Results

Iron deficiency enhanced EMMPRIN, MMP-9 production, and MMP-9 enzymatic activity in THP-1 derived macrophages and foam cells. Iron deficiency elicited the activation of NF-κB and p38 MAPK. By using the p38 inhibitor and NF-κB inhibitor, the study established that EMMPRIN and MMP-9 inductions by iron deficiency required the consecutive upstream activation of p38 MAPK and NF-κB. This pro-inflammatory action was not prevented by PPARγ agonist. Meanwhile, iron deficiency did not modulate PPARγ expression. Retinoid X receptor agonist suppressed the effects of iron deficiency on EMMPRIN , MMP-9, and NF-κB, but not on MAPK activation.

Conclusions

Iron deficiency enhances atheroma inflammation through p38 MAPK-NF-κB-EMMPRIN/MMP-9 pathway. Our findings provide a potential mechanism for the association of major bleeding with recurrent ACS and mortality in patients with ACS.     

MMP-2、MMP-9及EMMPRIN在子宫内膜异位症中的表达及临床意义

目的研究基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）、细胞外基质金属蛋白酶诱导因子（EMMPRIN）在子宫内膜异位症（EMs）的表达和意义。方法应用免疫组化二步法检测EMs患者异位内膜42例、在位内膜42例及正常内膜20例中的MMP-2、MMP-9、EMMPRIN的表达情况,并对它们的EMMPRIN、MMP-2、MMP-9蛋白表达水平进行相关性分析。结果 MMP-2、MMP-9、EMMPRIN在异位内膜组中阳性表达率分别为95.24%、92.86%和90.48%,显著高于在位内膜组、正常内膜组（P〈0.05）;而在位内膜组和正常内膜组差异无统计学意义（P〉0.05）。异位内膜组中,EMMPRIN分别和MMP-2,MMP-9呈正相关性（P〈0.01）。结论 MMP-2、MMP-9、EMMPRIN共同参与了子宫内膜异位症的发生发展;EMMPRIN可能通过促进MMP-2和MMP-9的合成与分泌发挥其作用。     

细胞外基质金属蛋白酶诱导因子、基质金属蛋白酶9及P38丝裂原活化蛋白激酶在子宫内膜异位症中的表达及相关性

目的探讨细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,EMMPRIN)、基质金属蛋白酶9(matrix metalloproteinases-9,MMP-9)和P38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)在子宫内膜异位症(endometriosis,EMs)中的表达及相关性。方法收集2008年5-12月重庆医科大学附属第一医院收治的EMs患者44例(异位内膜44例,在位内膜38例),对照组为同期非EMs患者在位内膜34例。应用免疫组化SABC三步法检测各组织中EMMPRIN、MMP-9、P38及磷酸化P385(p-P38)蛋白的表达,并对各蛋白表达水平进行相关性分析。结果 EMMPRIN、MMP-9、P38、p-P38蛋白在EMs异位内膜表达均高于EMs在位内膜组及对照组在位内膜,差异有统计学意义(P<0.05);Spearman相关分析显示,在EMs异位内膜中各蛋白表达呈正相关(P<0.05)。在EMs异位内膜组中各蛋白表达Ⅲ～Ⅳ期高于Ⅰ～Ⅱ期,差异均有统计学意义(P<0....