ADAM9在胰腺癌发生发展中的作用研究

金属蛋白酶解离素9(A Disintegrin and Metalloprotease 9,ADAM9)是ADAM家族中的一员,研究表明ADAM9能提高肿瘤细胞的侵袭与转移能力,和肿瘤发生、侵袭、转移有关.本研究旨在明确ADAM9在胰腺癌细胞中的作用,以期为胰腺癌治疗寻找新的靶点.     

ADAM9在乳腺癌中的表达及意义

目的探讨去整合素金属蛋白酶9(ADAM9)在乳腺癌中的表达及意义。方法采用逆转录多聚酶链聚合反应(RT-PCR)法和免疫组织化学方法检测乳腺癌组织、距离癌旁≥5cm的正常乳腺组织及腋窝淋巴结组织中ADAM9的表达,并分析ADAM9表达与乳腺癌临床病理特征之间的关系。结果 ADAM9mRNA在乳腺癌组织中的表达较正常乳腺组织(未检测到ADAM9mRNA的表达)明显增强。ADAM9蛋白表达阳性率在乳腺癌组织中明显高于正常乳腺组织(P0.05),其在乳腺癌转移淋巴结中的表达明显高于非转移淋巴结及相应原发灶中的表达(P0.05).乳腺癌组织中ADAM9蛋白表达阳性率与其淋巴结转移和组织学分级有关(P0.05).结论ADAM9在乳腺癌中表达上调,可能参与了乳腺癌发生、发展过程。     

解聚素-金属蛋白酶17在结肠直肠癌中的表达及其意义

目的:研究解聚素-金属蛋白酶17(a disintegrin and metalloprotease17,ADAM17)在结肠直肠癌中的表达及其临床意义.方法:收集80例手术切除的结肠直肠癌及60例正常结肠直肠黏膜组织,采用RT-PCR和免疫组化SP法检测ADAM17在结肠直肠癌及正常结肠直肠黏膜组织中的表达情况:分析ADAM17在结肠直肠癌组织中的表达和病人临床病理特征之间的关系.结果:结肠直肠癌组织的ADAM17基因表达高于正常结肠直肠黏膜组织(0.66±O.14比0.26±0.12,P<0.01),ADAM17蛋白在结肠直肠癌组织中之表达阳性率为79%,在正常结肠直肠黏膜组织中之表达阳性率为25%,差异具有统计学意义(P<0.01).ADAM17高表达与结肠直肠癌浸润深度及远处转移相关(P<0.05);与病人的年龄、性别、肿瘤部位、大小、分化程度及淋巴结转移无关.结论:ADAM17在结肠直肠癌组织中呈高表达,此情况与结肠直肠癌浸润深度及远处转移相关,可能成为一种新的判断结肠直肠癌病人预后的标志物.     

基质金属蛋白酶-9在临床上的应用

肿瘤细胞的侵袭和转移过程是一个复杂多步骤过程，其中金属蛋白酶-9起着一个重要作用。本文从它的结构、基因表达、检测、功能和金属蛋白酶抑制剂（TIMPs）的关系及在各肿瘤中的表达作一综述。     

基质金属蛋白酶和组织金属蛋白酶抑制剂在大肠癌中的表达

目的　研究基质金属蛋白酶 (MMPs)及组织金属蛋白酶抑制剂 (TIMPs)在大肠癌中的表达特点 ,与肿瘤发生发展的关系 ,以及在肿瘤治疗中的应用前景。　方法　采用RT PCR方法测定 2 8例大肠癌患者肿瘤组织和周围正常粘膜的基质金属蛋白酶 2 (MMP 2 )、膜型 1 基质金属蛋白酶 (MT1 MMP)、基质溶素 (MMP 7)、组织金属蛋白酶抑制剂 2 (TIMP 2 )、组织金属蛋白酶抑制剂 3(TIMP 3)的mRNA表达状况 ,并将其结果与临床及病理学资料进行统计学分析。　结果　 (1) 2 7例患者肿瘤组织中MMP 7mRNA表达阳性 ,MMP 2、MT1 MMP、TIMP 2和TIMP 3在肿瘤组织和正常粘膜中均有高表达 ;(2 )肿瘤组织中MMP 7mRNA的表达水平与大肠癌患者的Dukes′分期相关 (P <0 0 1) ;(3)淋巴结阳性患者的肿瘤组织TIMP 2表达水平为 (1 2 5± 0 46 )明显高于淋巴结阴性患者的 (0 75± 0 41) ,差异有显著性意义 (P <0 0 1) ;(4)大肠癌患者癌周正常粘膜TIMP 3mRNA表达随患者Duke′s分期的进展和肿瘤浸润深度的增加而降低 (P <0 0 1) ;(5 )TIMPs与MMPs之间无明显相关关系 (P >0 1)。　结论　MMP 7可望成为诊断大肠癌的敏感指标 ;人工诱导TIMP 2、TIMP 3或阻断MMP 7、MMP 2、MT1 MMP的表达可能抑制肿瘤的浸润和转移 ,成为肿瘤治疗的新途径。     

MMP-9和TIMP-1在膀胱癌中的表达及临床意义

目的：研究基质金属蛋白酶9（MMP-9）和组织型金属基质蛋白酶抑制因子1（TIMP-1）在膀胱移行细胞癌中的表达及其与肿瘤侵袭转移的关系。方法：采用免疫组织化学SP法检测56例膀胱移行细胞癌标本中MMP-9和TIMP-1的表达,并与肿瘤的临床及病理参数相比较。结果：在56例膀胱移行细胞癌中,MMP-9和TIMP-1的表达率分别为6．1．29％和57．14％,MMP-9表达率随肿瘤的临床分期、病理分级的升高而增加,TIMP—1表达率也呈上升趋势,两者间的表达仔在轻度正相关。结论：MMP-9和TIMP—1的相互作用对于膀胱移行细胞癌的浸润转移发挥了重要作用,MMP-9可能成为早期无创诊断膀胱肿瘤的生物学指标。     

解整合素金属蛋白酶9在胆管癌细胞中的表达及临床意义

解整合素金属蛋白酶(ADAM)是依赖锌离子的基质金属蛋白酶(MMP)超家族的一员[1],是一类含有解整合素和金属蛋白酶结构域的Ⅰ型跨膜蛋白.近年来有研究证实ADAM9蛋白在肝癌、乳腺癌、结肠癌、甲状腺癌、前列腺癌等恶性肿瘤中均有表达[2].本研究旨在探讨ADAM9在胆管癌细胞中的表达及临床意义. 一、材料和方法 1.材料:一抗为兔抗人ADAM9多克隆抗体(购自北京博奥森生物技术有限公司),免疫组织化学二抗试剂盒以及二氨基联苯胺(DAB)显色试剂盒均购白武汉博士德生物工程公司.     

金属蛋白酶抑制基因RECK和基质金属蛋白酶2、9在膀胱癌组织中的表达及意义

我们采用RT—PCR方法检测金属蛋白酶抑制基因RECK及基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）基因在43例膀胱癌（TCC）组织标本中的表达，探讨以上基因与TCC侵袭、转移等生物学行为的关系。现报告如下。     

大肠癌组织中基质金属蛋白酶-9、基质金属蛋白酶抑制剂-1的表达及对微血管生成和肿瘤转移的影响

目的　探讨基质金属蛋白酶 -9(MMP -9)与基质金属蛋白酶抑制剂 -1(TIMP -1)在大肠癌中的表达及其与血管生成 ,侵袭转移之间的关系。方法　采用免疫组化S -P法 ,检测 46例大肠癌组织中MMP -9,TIMP -1的表达和微血管密度之间的关系。结果　MMP -9的表达与肿瘤的分化程度、浸润深度、Dukes分期及淋巴结转移明显相关 (P <0 .0 5或P <0 .0 1) ;TIMP -1的表达与Dukes分期、淋巴结转移明显相关 ( P <0 .0 5或 P <0 .0 1) ;MMP -9表达阳性的大肠癌组织微血管密度 (MVD)值明显高于表达阴性者 (P <0 .0 1)。结论　MMP -9、TIMP -1的表达和微血管生成与大肠癌侵袭转移密切相关     

MMP-2、MMP-9在膀胱移行细胞癌中的表达及临床意义

目的 探讨基质金属蛋白酶2(MMP2),基质金属蛋白酶9(MMP9)在膀胱移行细胞癌中的表达及临床意义.方法 采用免疫组化SP法和高清晰度彩色医学图文分析系统对35例膀胱移行细胞癌和10例正常膀胱黏膜中MMP-2、MMP-9的表达进行检测分析.结果 MMP-2、MMP-9在膀胱移行细胞癌中呈高表达,且随分期、分级的增高而增高(p＜0.01).结论 MMP-2、MMP-9参与膀胱癌侵袭转移,在膀胱移行细胞癌中的表达对其预后判断具有参考价值.     

Enhanced Allograft Survival and Modulation of T-Cell Alloreactivity Induced by Inhibition of MMP/ADAM Enzymatic Activity

Recent studies have shown significantly increased expression of matrix metalloproteinases (MMP) and disintegrin-type metalloproteinases (ADAM) during allograft rejection. In this regard, our previous studies have demonstrated contrasting roles for MMP-2 and MMP-9 during allograft rejection: MMP-2-deficiency enhanced allograft survival while MMP-9-deficiency decreased allograft survival. The aim of this study was to determine the effect of broad-spectrum MMP/ADAM inhibition on the pathogenesis of allograft rejection. Toward this, heterotopic BALB/c cardiac allografts were transplanted into C57BL/6 recipients treated with MMP/ADAM inhibitors, GM6001 or doxycycline. Systemic MMP/ADAM inhibition significantly enhanced allograft survival. Functioning allografts recovered from MMP/ADAM inhibitor-treated recipients showed lower cellular infiltration and tissue remodeling than rejected allografts recovered from control recipients. In addition, decreased chemotaxis of CD4+ and CD8+ T cells, B cells and macrophages was observed in vitro in the presence of MMP/ADAM inhibitors. Enhanced T-cell alloreactivity was also observed ex vivo in MMP/ADAM inhibitor-treated recipients and in vitro in the presence of MMP/ADAM inhibitors. These observations were associated with enhanced cytokine, chemokine and growth factor production. These results indicate that MMPs and ADAMs play a critical role in the pathogenesis of allograft rejection and may represent novel therapeutic targets for the treatment and/or prevention of this disease.     

ADAM8 enhances osteoclast precursor fusion and osteoclast formation in vitro and in vivo

ADAM8 expression is increased in the interface tissue around a loosened hip prosthesis and in the pannus and synovium of patients with rheumatoid arthritis, but its potential role in these processes is unclear. ADAM8 stimulates osteoclast (OCL) formation, but the effects of overexpression or loss of expression of ADAM8 in vivo and the mechanisms responsible for the effects of ADAM8 on osteoclastogenesis are unknown. Therefore, to determine the effects of modulating ADAM expression, we generated tartrate‐resistant acid phosphatase (TRAP)–ADAM8 transgenic mice that overexpress ADAM8 in the OCL lineage and ADAM8 knockout (ADAM8 KO) mice. TRAP‐ADAM8 mice developed osteopenia and had increased numbers of OCL precursors that formed hypermultinucleated OCLs with an increased bone‐resorbing capacity per OCL. They also had an enhanced differentiation capacity, increased TRAF6 expression, and increased NF‐κB, Erk, and Akt signaling compared with wild‐type (WT) littermates. This increased bone‐resorbing capacity per OCL was associated with increased levels of p‐Pyk2 and p‐Src activation. In contrast, ADAM8 KO mice did not display a bone phenotype in vivo, but unlike WT littermates, they did not increase RANKL production, OCL formation, or calvarial fibrosis in response to tumor necrosis factor α (TNF‐α) in vivo. Since loss of ADAM8 does not inhibit basal bone remodeling but only blocks the enhanced OCL formation in response to TNF‐α, these results suggest that ADAM8 may be an attractive therapeutic target for preventing bone destruction associated with inflammatory disease. © 2011 American Society for Bone and Mineral Research.     

Circular RNA circ_0114876 regulates osteoarthritis through upregulating ADAM10 via targeting miR-1227-3p

BackgroundOsteoarthritis (OA) was a chronic degenerative joint disease. The dysregulation of circular RNAs (circRNAs) has been identified in OA progression. However, the function and regulation mechanism of circ_0114876 in OA remains largely unknown.MethodFirstly, we used LPS-treated C28/I2 cells as a cellular model of OA. Quantificational real-time polymerase chain reaction (qRT-PCR) was used to determine the expression levels of circ_0114876, miRNA-1227-3p, and ADAM10 in OA chondrocytes. Cell Counting Kit-8 (CCK8), 5-ethynyl-20-deoxyuridine (EdU) incorporation assays, flow cytometry, Enzyme-linked immunosorbent assay (ELISA) kit, and western blot were applied to confirm cell proliferation, apoptosis, inflammation, and extracellular matrix.of circ_0114876 in vitro. The interaction between circ_0114876 and its downstream target (miR-1227-3p) and mRNA target ADAM metallopeptidase domain 10 (ADAM10), was evaluated by luciferase assay and RNA immunoprecipitation (RIP) assay.ResultCirc_0114876 and ADAM10 were upregulated and miR-1227-3p was decreased in OA tissues and LPS-treated chondrocytes. Low expression of circ_0114876 promoted proliferation and inhibited apoptosis, inflammation, and extracellular matrix of the LPS-treated chondrocytes. Mechanistically, circ_0114876 functioned in human chondrocytes through targeting miR-1227-3p and ADAM10. Furthermore, miRNA-1227-3p inhibitor reversed the effect of circ_0114876 knockdown on the OA chondrocytes, and ADAM10 overexpression reversed the effect of miR-1227-3p mimic on the OA chondrocytes.ConclusionCirc_0114876 was increased in OA tissues and cells. Circ_0114876 facilitated the progression in the LPS-induced OA cell model via regulating the miR-1227-3p/ADAM10 axis. This study would provide a potentially effective therapeutic strategy for OA progression.     

A novel marker ADAM17 for clear cell renal cell carcinomas: Implication for patients’ prognosis

IntroductionA disintegrin and metalloproteinase-17 (ADAM17) plays an important role in biological activity in different cancers. Its expression and prognostic value have not been studied in clear cell renal cell carcinoma (cRCC). The objective of this study was to explore the prognostic value of ADAM17 in patients with cRCC.Materials and methodsA total of 131 patients with cRCC were studied. There were 90 men and 41 women, with an average age of 67 years (range: 34–93 y). There were 110 patients with localized disease and 21 patients with metastatic disease. The expression of ADAM17 was evaluated by immunohistochemistry with a monoclonal antibody. The follow-up varied from 4.2 to 184 months, with a median of 72 months for patients with localized disease. Kaplan-Meier with a log-rank test was performed to compare the progression-free survival after surgery. The univariate and multivariate analyses were performed to examine the significance of ADAM17 expression for the patient’s progression-free survival.ResultsThe ADAM17 expression was found in 109/131 tumors. The ADAM17 expression was found in 20/21 metastatic tumors and in 89/110 localized tumors. Regarding patients with localized tumors, 31 patients experienced a recurrence or death during follow-up. The Kaplan-Meier analysis revealed that the high expression of ADAM17 was associated with a reduced progression-free survival (P = 0.005). The univariate logistic regression analysis indicated that the high expression of ADAM17 was associated with the disease progression (hazard ratio = 2.826; 95% CI: 1.324–6.034; P = 0.007). The high expression of ADAM17 remained a significant factor for decreased progression-free survival in multivariate analysis.ConclusionADAM17 was frequently expressed in cRCC. The high expression of ADAM17 was correlated with a worse outcome for patients with cRCC. ADAM17 is a new biomarker for the management of patients with cRCC.     

The ADAM17–amphiregulin–EGFR Axis in Mammary Development and Cancer

In order to fulfill its function of producing and delivering sufficient milk to newborn mammalian offspring, the mammary gland first has to form an extensive ductal network. As in all phases of mammary development, hormonal cues elicit local intra- and inter-cellular signaling cascades that regulate ductal growth and differentiation. Among other things, ductal development requires the epidermal growth factor receptor (EGFR), its ligand amphiregulin (AREG), and the transmembrane metalloproteinase ADAM17, which can cleave and release AREG from the cell surface so that it may interact with its receptor. Tissue recombination and transplantation studies demonstrate that EGFR phosphorylation and ductal development proceed only when ADAM17 and AREG are expressed on mammary epithelial cells and EGFR is present on stromal cells, and that local administration of soluble AREG can rescue the development of ADAM17-deficient transplants. Thus proper mammary morphogenesis requires the ADAM17-mediated release of AREG from ductal epithelial cells, the subsequent activation of EGFR on stromal cells, and EGFR-dependent stromal responses that in return elicit a new set of epithelial responses, all culminating in the formation of a fully functional ductal tree. This, however, raises new issues concerning what may act upstream, downstream or in parallel with the ADAM17-AREG-EGFR axis, how it may become hijacked or corrupted during the onset and evolution of cancer, and how such ill effects may be confronted.     

男性不育症患者精液ADAM基因表达的临床意义

目的探讨ADAM(去整合素和金属蛋白酶区家族)基因与不明原因不育症之间的关系。方法应用RT-PCR方法检测30例正常生育男性和30例不明原因不育症患者精液中ADAM1、ADAM2、ADAM3、ADAM32基因的 mRNA表达。结果正常组30例中4种基因 mRNA均有表达,不育组30例中有1例ADAM1、2、32呈阴性表达,1例ADAM2、32呈阴性表达,ADAM1和ADAM3呈阴性表达各1例,26例4种基因均呈阳性表达。结论ADAM1、2、3、32基因缺失或不表达可能是导致不育的候选基因之一。     

ADAM12-S stimulates bone growth in transgenic mice by modulating chondrocyte proliferation and maturation.

ADAM12-S transgenic mice exhibit a pronounced increase in the length of bones, such as femur, tibia, and vertebrae. The effect of ADAM12-S on longitudinal bone growth involves the modulation of chondrocyte proliferation and maturation, likely through proteolytic activities and altered cell-extracellular matrix interactions in the growth plate. INTRODUCTION: The disintegrin and metalloprotease ADAM12 is expressed in both osteoblasts and osteoclasts, suggesting a regulatory role of ADAM12 in bone. However, thus far, no in vivo function of ADAM12 in the skeleton has been reported. MATERIALS AND METHODS: Transgenic mice expressing the secreted form of human ADAM12, ADAM12-S, or a truncated metalloprotease-deficient form of ADAM12-S in the circulation were used to study the effects of ADAM12 on the skeleton. In addition, murine chondrocyte cultures were used to study the effect of ADAM12-S on cell-extracellular matrix interactions. RESULTS: ADAM12-S transgenic mice exhibit increased longitudinal bone growth. The increased bone length is progressive and age dependent, with a maximum increase of 17% seen in the femur from 6-month-old transgenic mice. The effect is gene dose dependent, being more pronounced in mice expressing higher levels of the transgene than in a lower-expressing line. Histological analysis revealed no alterations in the growth plate organization, but mean growth plate width was increased. Both the cellular incorporation of bromodeoxyuridine and the width of the collagen type X-positive hypertrophic zone were increased in the growth plate of ADAM12-S transgenic mice. Importantly, mice expressing a truncated form of ADAM12-S that lacked the pro- and metalloprotease domains showed no alterations in bone length, suggesting that protease activity is required for the ADAM12-S effect. In vitro studies showed that ADAM12-S inhibits chondrocyte adhesion to fibronectin and collagen type II. CONCLUSIONS: ADAM12-S stimulates bone growth in mice by modulating chondrocyte proliferation and maturation through mechanisms probably involving both metalloprotease and adhesion activities.     

ADAM gene expression and regulation during human osteoclast formation

In this study, we identified the expression and the regulation of ADAM members (a disintegrin and metalloprotease) at both gene and protein levels during human osteoclast differentiation and activity. Human peripheral blood monocytes (HPBMC) treated with M-CSF and RANKL were used as an in vitro fusion model. In parallel, we used human osteoclastoma (OCL) tumor as a source of mature osteoclasts, and human osteoblastic cells as a control representing nonfusing and non-resorbing bone cells. RT-PCR using ADAM-specific primers enabled us to identify the expression of ADAM 8, 9, 10, 15, 17, and 28 in both osteoclasts and osteoblasts. Using primers specific for each ADAM 12 isoform (L and S), we observed a strong signal for both forms (ADAM 12L and ADAM 12S) in osteoblastic cells, while only ADAM 12S was detectable in HPBMC-derived osteoclasts and osteoclastoma. Gene regulation was studied using real-time PCR analysis performed during HPBMC differentiation; this showed a progressive increase of ADAM 12 mRNA level from day 1 to 8 of the culture, while at around day 9, ADAM 12 mRNA level decreased 2-fold. We also showed that ADAM 8, ADAM 17, and ADAM 28 decreased according to the stage of HPBMC differentiation or fusion. ADAM 10 was unaltered during cell fusion. However, confocal immunolocalization showed that ADAM 10 protein re-localized from the nuclei and cytoplasm to the plasma membrane during culture and to the ruffled border in resorbing cells. The same re-localization process was observed using an ADAM 12S-specific antibody during HPBMC differentiation. Between days 12 and 14, ADAM 12 co-localized with the F-actin ring, and at day 15, a strong signal was also present in ruffled border or sealing zone area of osteoclasts. Our results describe the expression and regulation of various ADAMs in human bone cells and the selective expression of ADAM 12L in osteoblasts. Our gene regulation and protein localization studies suggest a function for ADAM 10 and ADAM 12S in the formation of osteoclasts from HPBMC and resorption activity.     

ADAM10在小鼠颅面部膜内成骨过程中的表达

目的观察解聚素样金属蛋白酶10(A disintegrin and metalloprotease 10,ADAM10)在小鼠颅面部骨骼膜发育过程中的表达变化。方法以野生型C57BL/6小鼠为实验对象,阿辛蓝-茜素红染色显示小鼠膜内成骨区域,结合免疫组织荧光,检测ADAM10在骨组织中的表达。三标免疫荧光观察ADAM10在MC3T3细胞系中亚细胞定位。蛋白免疫印迹检测ADAM10在小鼠出生前后的表达量变化。结果组织阿辛蓝茜素红染色显示小鼠前颅骨、鼻旁软骨、上颌骨、腭板和下颌骨成骨活跃,并结合免疫组织荧光检测,发现ADAM10在小鼠前颌骨、鼻旁软骨、上颌骨、腭板和下颌骨广泛表达,且主要表达在成骨活跃的区域。MC3T3细胞三标免疫荧光检测进一步定位ADAM10广泛分布在胞浆中,在质膜和细胞核附近表达高;其胞核附近的高表达信号与高尔基体共标,提示其可能在高尔基体中加工后至细胞膜发挥功能。同时,蛋白免疫印迹结果证实,ADAM10在小鼠出生前后表达高,成年后表达明显降低。结论 ADAM10广泛表达于小鼠早期颅面部膜内成骨活跃区域,提示其可能调控小鼠颅面部膜内成骨过程。     

ADAM19 expression in human nephrogenesis and renal disease: associations with clinical and structural deterioration

ADAM19, an enzyme from the ADAM (a disintegrin and metalloproteinase) family, is involved in various cell-cell and cell-matrix interactions. It can cleave epidermal growth factor (EGF)-like growth factors, such as heparin-binding (HB)-EGF and neuregulin (NRG), from the cell membrane. ADAM-mediated EGF receptor activation is crucial in the development of renal pathology. Based on these data, we studied ADAM19 in human nephrogenesis and renal disease. We collected 20 fetal kidneys and 56 biopsies from patients with various renal diseases. The unaffected part of kidneys from eight patients with renal cell carcinoma served as control. RNA in situ hybridization revealed widespread ADAM19 mRNA expression in the nephrogenic zone of human fetal kidneys. Normal human kidneys showed constitutive ADAM19 expression in distal tubules and endothelial cells, whereas proximal tubules were negative. In renal disease, ADAM19 was de novo expressed in proximal tubules and glomerular mesangium and upregulated in distal tubules and endothelial cells. ADAM19 colocalized with tubular and interstitial NRG, however, not with HB-EGF. Independent of renal disorder, mesangial ADAM19 expression was associated with glomerular damage as assessed by mesangial matrix expansion, focal glomerulosclerosis, and glomerular macrophage influx (all P<0.001). ADAM19 in proximal tubules and in peritubular capillaries was associated with interstitial fibrosis (P<0.05). Finally, increasing tubular ADAM19 was associated with declining renal function (P<0.05). The abundant ADAM19 expression during nephrogenesis points to a role in growth promotion and regulation. The high ADAM19 expression in renal disease suggests involvement in profibrotic and proinflammatory processes leading to renal deterioration.