结直肠癌组织中Twist、MMP-2和MMP-9的表达及意义

目的 探讨Twist与MMP-2、MMP-9在结直肠癌中表达的相关性.方法 采用免疫组化SP法检测48例结直肠癌组织和24例癌旁5 cm以上正常组织中Twist、MMP-2、MMP-9的表达情况.结果 在结直肠癌组织中,Twist、MMP-2、MMP-9的阳性表达率分别为47.92%、54.17%,62.50%,均显著高于癌旁正常组织中的12.50%、20.83%,25.00%,(P＜0.05).Twist、MMP-2、MMP-9与肿瘤的分化程度、浸润深度、淋巴结转移有关(P＜0.05).Twist的表达与MMP-2、MMP-9的表达呈正相关.MMP-2的表达与MMP-9的表达呈正相关.结论 Twist、MMP-2、MMP-9在肿瘤细胞发生浸润转移中起了协同作用,可作为评估结直肠癌转移及预后的参考指标.     

MMP-9在胃腺癌中的表达意义

目的 探讨MMP-9在胃腺癌中的表达意义.方法 采用免疫组化S-P法检测10例正常胃黏膜上皮、20例癌旁组织、60例胃腺癌组织中MMP-9的表达,并分析其表达意义及与胃癌临床病理特征的关系.结果 在正常胃黏膜上皮、癌旁组织、胃腺癌组织中,MMP-9的表达阳性率在胃腺癌组织中显著高于正常胃黏膜上皮及癌旁组织中(P＜0.01),在癌旁组织中显著较高于正常组织中(P＜0.05).MMP-9的表达阳性率在低分化癌组显著高于中分化癌组(P＜0.05),在浸润肌层及浸润浆膜层组显著高于浸润黏膜下层组(P＜0.05),在淋巴结转移组显著高于无淋巴结转移组(P＜0.05).MMP-9的表达阳性率与患者性别、年龄、肿瘤体积等无关(P>0.05).结论 MMP-9的表达增强可能与胃癌的转移侵袭有关;MMP-9可能促进了胃癌的发生发展.     

MMP-1和TIMP-1在人胃癌组织中的表达及临床意义

目的 探讨基质金属蛋白酶1(MMP-1)和金属蛋白酶1组织抑制因子(TIMP-1)基因与胃癌临床生物学行为的关系.方法 采用逆转录-聚合酶链反应(RT-PCR)检测60例胃癌组织及其相应癌旁正常胃黏膜组织中MMP-1和TIMP-1 mRNA的表达.结果 60例胃癌组织中MMP-1表达水平明显高于癌旁正常组织,TIMP-1表达水平明显低于癌旁正常组织(P<0.05).胃癌组织中,MMP-1 mRNA的相对表达强度、未分化癌明显高于高、中分化癌,Ⅲ～Ⅳ期明显高于Ⅰ～Ⅱ期,伴淋巴结转移者明显高于无淋巴结转移者(P<0.05);TIMP-1 mRNA的相对表达强度低、未分化癌则明显低于高、中分化癌,Ⅲ～Ⅳ期明显低于Ⅰ～Ⅱ期,伴淋巴结转移者明显低于无淋巴结转移者(P<0.05).但MMP-1、TIMP-1表达与患者年龄、性别、肿瘤病理类型及肿瘤大小和部位等临床参数无关(P>0.05).MMP-I与TIMP-1表达呈负相关(r=-0.513,P<0.05).结论 胃癌中MMP-1、TIMP-1表达与胃癌分化程度、TNM分期及淋巴结转移有关.     

MMP-13在甲状腺癌中的表达及其临床意义

目的探讨MMP-13在甲状腺癌组织中的表达及其与疾病分期的关系。方法采用免疫组化法检测62例甲状腺癌患者的癌组织以及癌旁正常组织中MMP-13蛋白的表达水平,分析MMP-13表达与甲状腺癌临床分期的关系。结果甲状腺癌组织中MMP-13蛋白表达的阳性率为77.4%(48/62),显著高于癌旁正常组织的8.1%(5/62)(P<0.05);晚期(Ⅲ～Ⅳ期)甲状腺癌患者癌组织中MMP-13蛋白表达的阳性率为94.7%(18/19),显著高于早期(Ⅰ～Ⅱ期)甲状腺癌患者的77.4%(48/62)(P<0.05)。结论甲状腺癌组织中MMP-13蛋白的表达水平升高,MMP-13的高表达与甲状腺癌的疾病进展密切相关。     

MMP-9 TIMP-1在肝细胞癌中的表达及意义

目的探讨基质金属蛋白酶-9（MMP-9）及其金属蛋白酶组织抑制剂-1（TIMP—1）在肝细胞癌（HCC）中表达及其临床病理意义。方法通过免疫组化SABC法检测42例肝细胞癌中MMP-9和TIMP—1的表达情况。结果MMP-9、TIMP—1在HCC与癌旁组织的阳性表达率相比,有显著差异（P〈0、05）。MMP-9在有癌栓组、包膜完整组的阳性率与无癌栓组、包膜不完整组阳性率相比,具有显著性差异（P〈0．05）。MMP-9和TIMP—1的在肝细胞癌（HCC）中阳性率表达呈正相关。结论MMP-9促进肝细胞癌浸袭转移。体内TIMP—1的增高程度不足以抑制MMP-9的活性。     

结肠癌组织AQP1与MMP-2、MMP-9、TIMP-1表达的关系及意义

目的 检测水通道蛋白1(AQP1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制因子-1(TIMP-1)在结肠癌组织中的表达,并分析这些蛋白质之间的关系,探讨其意义.方法 对54例结肠癌组织和40例癌旁组织石蜡标本进行免疫组化染色,检测AQP1与ICAM-1、MMP-2、MMP-9、TIMP-1在两种组织中的表达情况.结果 结肠癌组织中AQP1与ICAM-1、MMP-2、MMP-9、TIMP-1阳性表达率分别为75.93%、64.81%、77.78%、46.30%,而癌旁组织中阳性率则分别为22.50%、37.50%、50.00%、70.00%.AQP1、MMP-2、MMP-9在肿瘤组织中表达强度高于癌旁组织,而TIMP-1在结肠癌组织中表达则低于癌旁组织(P<0.05).肿瘤组织中AQP1与MMP-2、MMP-9表达呈正相关性(r值分别为0.4030,0.3874,P<0.05),MMP-2、MMP-9与TIMP-1表达呈负相关(r值分别为-0.3176、-0.4728,P<0.05).结论 结肠癌组织中AQP1表达增强,AQP1可能通过调节MMP-2、MMP-9表达而促进结肠癌的侵袭转移.     

胃癌组织中CD54与MMP-9的表达及其相关性研究

目的 探讨CD54、MMP-9在胃癌原发灶中的表达情况及2者与肿瘤浸润转移、肿瘤组织学类型之间的相关性.方法 用免疫组织化学染色方法 分别检测CD54和M胆9在46例胃癌原发灶、癌旁3.0 cm、癌旁10.0 cm及正常组织(外伤或病理证实为良性溃疡)内的表达情况.结果 CD54在胃癌原发灶内的阳性表达率为67.39%(31/46),癌旁3.0 cm、癌旁10.0 am和正常组织中的阳性表达率分别为54.35%(25/46)、8.70%(4/46)、0.00%(0/12),MMP-9在胃癌组织中阳性表达率为82.61%(38/46),癌旁3.0 cm、癌旁10.0 cm和正常组织阳性表达率分别为65.22%(30/46)、10.87%(5/46)和0.00%(0/12).CD54与MMP-9的表达在胃癌组织中明显升高,呈正相关(P<0.05).结论 CD54和MMP-9在胃癌原发灶组织中均呈高表达,并两者正相关,并可能对肿瘤进展、恶性程度的判断以及发生机制具有重要意义,但两者在共同作用促进肿瘤进展的具体机制还有结于进一步研究.     

PCNA和MMP-9在食管鳞癌中的表达及其意义

目的分析PCNA,MMP-9在食管鳞癌中的表达,探讨其在食管鳞癌发生发展中的意义。方法应用免疫组化方法检测PCNA,MMP-9在52例食管鳞癌和30例癌旁组织中的表达情况,并结合临床病理资料进行分析。结果PCNA,MMP-9蛋白在食管鳞癌组织中的表达率高于癌旁组织,差异具有统计学意义(P<0.05)。PCNA的表达与食管鳞癌的分化程度,淋巴结转移有关;MMP-9的表达与食管鳞癌分化程度,肿瘤浸润程度,淋巴结转移及临床分期相关;PCNA和MMP-9表达呈正相关(P<0.05)。结论食管鳞癌组织中PCNA,MMP-9的高表达参与了肿瘤的发生发展,联合检测有助于食管鳞癌恶性生物学行为的判断和病程的评估。     

MMP-2和PTEN蛋白在肝癌组织中的表达及临床意义

目的 探讨肝癌组织中MMP-2和PTEN的表达及两者与肝癌侵袭性的关系.方法 用免疫组化SP法检测61例肝癌组织和17例正常肝组织中MMP-2和PTEN蛋白质的表达,并分析两者与肝癌临床病理特征,侵袭性的关系.结果 HCC中PTEN阳性表达率明显弱于正常肝组织,而MMP-2阳性表达率明显高于正常肝组织,且两者呈负相关;PTEN蛋白阳性表达率与HCC组织分化程度、侵袭性有关(P＜0.05),而与性别、术前AFP水平无关(P＞0.05);MMP-2阳性表达率与HCC组织侵袭性有关(P＞0.05),而与HCC组织分化程度、性别、术前AFP水平无关(P＜0.05).结论 PTEN、MMP-2的表达一定程度上反映HCC侵袭性强弱;PTEN缺失可引起MMP-2表达增加,在HCC侵袭、转移中发挥作用.     

膀胱癌中MMP-2及TIMP-2的表达与病理分期和预后的相关性

目的探讨金属蛋白酶2(MMP-2)及其抑制因子(TIMP-2)在膀胱癌组织中的表达与病理学分级、分期和患者预后的相关性。方法采用免疫组化SP法,检测MMP-2及TIMP-2在40例膀胱癌和癌旁组织中的表达;术后随访2年。结果膀胱移行细胞癌(BTCC)组织中MMP-2表达阳性率为42.5%(17/40);TIMP-2表达阳性率为32.5%(13/40)。癌旁组织中MMP-2表达阳性率为25%(10/40),TIMP-2表达阳性率为20%(8/40)。对32例膀胱部分切除术患者随访2年,结果 12例膀胱癌复发。复发12例中,BTCC组织MMP-2表达阳率为91.6%(11/12),TIMP-2表达阳性率为33.3%(4/12);癌旁组织MMP-2表达阳率为66.7%(8/12),TIMP-2表达阳性率为16.7%(2/12)。结论 MMP-2和TIMP-2的表达阳性率BTCC显著高于癌旁组织(P<0.01)。BTCC组织中MMP-2的阳性表达与临床分级呈正相关(P<0.01);TIMP-2的阳性表达与临床分级相关不显著(P>0.05)。膀胱部分切除术患者高MMP-2阳性表达率和/或低TIMP-2阳性表达率以及癌旁组织MMP-2阳性表达与术后膀胱癌复发密切相关。高MMP-2阳性表达率和/或低TIMP-2阳性表达率表示膀胱癌浸润及转移能力增强;癌旁组织MMP-2阳性表达的膀胱癌具有恶性程度高、易复发的特点;提示手术时尽可能保证足够的切除范围。     

MMP-1、MMP-13在大鼠骨关节炎软骨中的表达

目的观察MMP-1和MMP-13在大鼠前交叉韧带切断加内侧半月板切除的骨关节炎模型中的表达,探讨MMP-1和MMP-13表达与骨关节炎(OA)进程的关系。方法将25只SD大鼠右膝行前交叉韧带切断加内侧半月板切除术(ACLT+MMx),对侧作为空白对照,分别于术后1周、2周、4周、6周、10周处死,每次随机处死5只,另取5只SD大鼠仅行右膝关节切开术作为假手术组,于术后10周处死,对膝关节软骨标本进行大体和组织学切片观察及评分,用ELISA法检测MMP-1、MMP-13在关节软骨中的表达。结果 ACLT+MMx组Mankin评分由1周的1.0分到4周的6.2分发展到10周的13.6分,与假手术组及空白对照组相比有显著差异(P<0.01)。ACLT+MMx组MMP-13的表达在OA早期增高,中重度期后出现明显下降趋势(P<0.01);MMP-1的表达则随着病程持续升高,与病变严重程度呈正相关性(P<0.01)。假手术组及空白对照组软骨细胞几乎不表达MMP-1及MMP-13。结论 MMPs在OA的发病机制中有重要作用,MMPs各亚型在OA进程中的表达各不相同。     

Arylsulphonamide hydroxamic acids as potent inhibitors of MMP-13

Pfizer has disclosed a series of phenoxyphenyl sulphonamide hydroxamic acids, containing Cα gem-disubstitution and a novel N-ethylcarboxylate moiety, which are potent inhibitors of matrix metalloproteinase-13 (MMP-13), an enzyme which has been implicated in such disease states as cancer and arthritis. The compounds are significantly selective (300-1000 fold) for MMP-13 versus MMP-1, the inhibition of which is believed to be associated with clinical side effects with previous broad spectrum MMP inhibitors. The Pfizer compounds are equally or more selective than several current clinical candidates and may have favourable pharmacodynamic profiles.     

Design strategies for the identification of MMP-13 and Tace inhibitors

Inhibitors of matrix metalloprotease (MMP)-13 and tumor necrosis factor-alpha converting enzyme (TACE) have been highly sought as potential therapeutic agents for the treatment of osteoarthritis and rheumatoid arthritis, respectively. This review focuses on the published literature on these inhibitors from 2001 to mid-2003. Significant advances have been reported in the design and synthesis of potent and selective inhibitors of MMP-13 using hydroxamic acid and non-hydroxamate zinc chelators on a variety of scaffolds. TACE inhibitors based on variations of known MMP inhibitors scaffolds and novel designs have been reported. Selectivity profiles for these inhibitors range from broad-spectrum to TACE-specific. Future clinical studies on these and other inhibitors will determine which MMP, or set of MMPs, must be inhibited for efficacy and long-term safety.     

Joint diseases and matrix metalloproteinases: a role for MMP-13

The role of matrix metalloproteinases in disease has been investigated over the last two decades. A focus on this family of proteases is particularly emphasized in two major arthritides in humans, osteoarthritis and rheumatoid arthritis. Early work described the presence of multiple MMP family members in the joint of the disease state and recent advances in the development of new knockout mice and disease models have allowed investigators to directly test the role of the MMP proteases in arthritis. MMP-13 is expressed by chondrocytes and synovial cells in human OA and RA and is thought to play a critical role in cartilage destruction. The recent development of an MMP-13 knockout mouse has documented the important role for this enzyme in cartilage formation and further studies under disease conditions promise to reveal the function of this enzyme in disease pathology. This review describes a body of research that supports the development of novel selective MMP-13 inhibitors with the hope of developing these compounds in clinical trials for the treatment of arthritis.     

N-O-Isopropyl sulfonamido-based hydroxamates: Kinetic characterisation of a series of MMP-12/MMP-13 dual target inhibitors

Matrix metalloproteinases (MMPs) are a family of zinc dependent endopeptidases known to play key roles in extracellular matrix (ECM) breakdown disorders, such as the two main forms of arthritis, rheumatoid arthritis (RA) and osteoarthritis (OA). MMP-13 (collagenase 3) is the leading MMP involved in cartilage degradation through its particular ability to cleave type-II collagen and as such plays a pivotal role in the pathogenesis of these diseases. Here we report the kinetic characterisation of N-O-isopropyl sulfonamido-based hydroxamates, potent inhibitors of MMP-13 and MMP-12, bearing different P1 and P1' substituents. One of these compounds proved to be a potent (4≤K(i)≤5nM) slow-binding inhibitor towards MMP-13 and MMP-12, with very favourable low association (10(4)M(-1)s(-1)) and dissociation constants (10(-4)s(-1)). Moreover, this compound exhibited a good selectivity for MMP-13 and MMP-12 over MMP-1, MMP-3, MMP-7, MMP-8 and, even to a minor extent, MMP-2. A molecular-docking study carried out using the experimentally-derived X-ray crystal structure of MMP-12 (PDB ID: 3F17) revealed critical hydrogen bonding of the hydroxamate and the sulfonamide moieties with key active site residues. Since also MMP-12 is involved in RA, this MMP-13/MMP-12 dual target inhibitor could be a valid candidate for the treatment of this pathology.     

Hydrogen sulfide suppresses the expression of MMP-8, MMP-13, and TIMP-1 in left ventricles of rats with cardiac volume overload

Aim:

To study the effects of hydrogen sulfide (H₂S) on the left ventricular expression of MMP-8, MMP-13, and TIMP-1 in a rat model of congenital heart disease.

Methods:

Male SD rats underwent abdominal aorta-inferior vena cava shunt operation. H₂S donor NaHS (56 μmol·kg⁻¹·d⁻¹, ip) was injected from the next day for 8 weeks. At 8 weeks, the hemodynamic parameters, including the left ventricular systolic pressure (LVSP), the left ventricular peak rate of contraction and relaxation (LV±dp/dt_max) and the left ventricular end diastolic pressure (LVEDP) were measured. The left ventricular tissues were dissected out, and hydroxyproline and collagen I contents were detected with ELISA. The expression of MMP-8, MMP-13, and a tissue inhibitor of metalloproteinase-1 (TIMP-1) in the tissues was measured using real-time PCR, Western blotting, and immunohistochemistry, respectively.

Results:

The shunt operation markedly reduced LVSP and LV±dp/dt_max, increased LVEDP, hydroxyproline and collagen I contents, as well as the mRNA and protein levels of MMP-8, MMP-13, and TIMP-1 in the left ventricles. Chronic treatment of the shunt operation rats with NaHS effectively prevented the abnormalities in the hemodynamic parameters, hydroxyproline and collagen I contents, and the mRNA and protein levels of MMP-13 and TIMP-1 in the left ventricles. NaHS also prevented the increase of MMP-8 protein expression, but did not affect the increase of mRNA level of MMP-8 in the shunt operation rats.

Conclusion:

H₂S suppresses protein and mRNA expression of MMP-8, MMP-13, and TIMP-1 in rats with cardiac volume overload, which may be contributed to the amelioration of ventricular structural remodeling and cardiac function.     

New hope for the treatment of osteoarthritis through selective inhibition of MMP-13

Osteoarthritis (OA) is the leading cause of joint pain and disability in middle-aged and elderly patients, and is characterized by progressive loss of articular cartilage that eventually leads to a complex process involving degradation of various components of the cartilage matrix, chief among them are the cartilage-specific type II collagen (CII) and aggrecan. While the loss of aggrecan is thought to be an early and reversible process, degradation of CII is considered to be irreversible and a key step in the loss of structural and functional integrity of cartilage. Among the various matrix metalloproteinases (MMPs), MMP-13 is specifically expressed in the cartilage of human OA patients and is not present in normal adult cartilage. It is the major collagenase in OA cartilage and has the highest activity against CII. However, the clinical utility of broad-spectrum MMP inhibitors developed for treatment of OA has been restricted by dose- and duration-dependent musculoskeletal side effects in humans. Consequently, selectively inhibiting the MMP-13 would seem to be an attractive therapeutic objective. This review mainly focuses on selective MMP-13 inhibitors development in terms of OA since the late 90s, in terms of synthetic compounds of low molecular mass incorporating specific zinc-binding groups, non-zinc-binding groups. In addition, dual inhibitors of MMP-13 and aggrecanase are also reviewed. Special emphasis is placed on logistic concerns for lead compound search as well as the structure-activity relationship (SAR) in this field. Through these methods, new hope is emerging for the treatment of OA through selective inhibition of MMP-13.     

Bisphosphonates inhibit stromelysin-1 (MMP-3), matrix metalloelastase (MMP-12), collagenase-3 (MMP-13) and enamelysin (MMP-20), but not urokinase-type plasminogen activator, and diminish invasion and migration of human malignant and endothelial cell lines

Bisphosphonates (clodronate, alendronate, pamidronate and zoledronate) at therapeutically attainable non-cytotoxic concentrations inhibited MMP-3, -12, -13 and -20 as well as MMP-1, -2, -8 and -9, but not urokinase-type plasminogen activator (uPA), a serine proteinase and a pro-MMP activator. Dose-dependent inhibition was shown by three independent MMP assays. The inhibition was reduced in the presence of an increased concentration of Ca(2+) when compared to physiologic Ca(2+) concentration. Alendronate inhibited the in vitro invasion (Matrigel) of human HT1080 fibrosarcoma and C8161 melanoma cells, and the random migration of these malignant and endothelial cell lines capable of expressing MMPs and uPA. The concentration of alendronate required to inhibit 50% of the activity (IC(50)=40-70 microM) of MMPs corresponded to the IC(50) of down-regulation of in vitro invasion and migration. The ability of bisphosphonates to down-regulate the in vitro invasion and random migration was comparable or slightly better in relation to the selective gelatinase inhibitor CTTHWGFTLC peptide. Alendronate but not CTTHWGFTLC peptide promoted the adhesion of HT1080 fibrosarcoma and C8161 melanoma cell lines on fibronectin. Bisphosphonates are broad-spectrum MMP inhibitors and this inhibition involves cation chelation. Bisphosphonates further exert antimetastatic, anti-invasive and cell adhesion-promoting properties, which may prevent metastases not only into hard tissues but also to soft tissues.     

Discovery and evaluation of a non-Zn chelating, selective matrix metalloproteinase 13 (MMP-13) inhibitor for potential intra-articular treatment of osteoarthritis

Osteoarthritis (OA) is a nonsystemic disease for which no oral or parenteral disease-modifying osteoarthritic drug (DMOAD) is currently available. Matrix metalloproteinase 13 (MMP-13) has attracted attention as a target with disease-modifying potential because of its major role in tissue destruction associated with OA. Being localized to one or a few joints, OA is amenable to intra-articular (IA) therapy, which has distinct advantages over oral therapies in terms of increasing therapeutic index, by maximizing drug delivery to cartilage and minimizing systemic exposure. Here we report on the synthesis and biological evaluation of a non-zinc binding MMP-13 selective inhibitor, 4-methyl-1-(S)-({5-[(3-oxo-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethyl)carbamoyl]pyrazolo[1,5-a]pyrimidine-7-carbonyl}amino)indan-5-carboxylic acid (1), that is uniquely suited as a potential IA-DMOAD: it has long durability in the joint, penetrates cartilage effectively, exhibits nearly no detectable systemic exposure, and has remarkable efficacy.     

咀嚼压力增强对大鼠剩余牙槽嵴中MMP-13表达的影响

目的观察咀嚼压力增强对大鼠剩余牙槽嵴中MMP-13表达的变化。方法建立大鼠剩余牙槽嵴创伤性吸收的动物实验模型,于3d、7d、14d、21d、56d后取材,采用SP免疫组织化学方法进行染色,观察MMP-13表达的变化。结果实验组牙槽骨吸收,MMP-13呈强阳性表达,且这种变化随着观察时间的变化而变化,在7d、14d时最明显,与对照组比较有统计学意义（p〈0．05）;而在28d、56d时二者差别不大。结论咀嚼压力增强时,剩余牙槽嵴中MMP-13表达的量增多,且其含量的变化与压力大小有关,力值越大,表达越强。