Ursolic acid-induced down-regulation of MMP-9 gene is mediated through the nuclear translocation of glucocorticoid receptor in HT1080 human fibrosarcoma cells

Cha, Hee-Jae,Park, Moon-Taek,Chung, Hae-Young,Kim, Nam-Deuk,Sato, Hiroshi,Seiki, Motoharu,Kim, Kyu-Won 부산대학교 유전공학연구소 1998 분자생물학 연구보 Vol.14 No.-

We have previously reported that ursolic acid, a pentacyclic triterpene acid, inhibited the invasion of HTI1080 human fibrosacrcoma cells by reducing the expression of matrix metalloproteinase-9. Since the chemical structure of ursolic acid is very similar to that of dexamethasone, a synthetic glucocorticoid, we investigated whether ursolic acid acts through the glucocorticoid receptor. The expression of matrix metalloproteinase-9 is thought to be regulated similary with matrix metalloproteinase-1 and matrix metalloproteinase-3 as containing common 2-0-teradecanoylphorbol-acetate responsible region, where AP-1 proteins can bind. Dexamethasone has been studied to respress the 2-0-teradecanoylphorbol-acetate-induced expression of matrix metalloproteinase-1 and matrix metalloproteinase-3 through a glucocorticoid receptor-mediated manner. In Northern biot analysis, we found that ursolic acid reduced the expression of matrix metalloproteinase-1 and matrix metalloproteinase-3 induced by 2-0-teradecanoylphorbol-acetate. Similarly, ursolic acid down-regulated 2-0-teradecanoylphorbol-acetate-induction of matrix metalloproteinase-9 gene in the same manner of dexamethasone. RU486, a potent glucocorticoid receptor antagonist, was used for identifying that ursolic acid-induced down-regulation of matrix metalloproteinase-9 expression is mediated by its binding to glucocorticoid receptor. The effect of ursolic acid on the matrix metalloproteinase-9 expression was blocked by RU486, suggesting that ursolic acid acts via a glucocorticoid receptor in the regulation of matrix metalloproteinase-9. Western blot analysis and immunocytochemistry showed that urslic acid increased glucocorticoid receptor fraction in the muscleus, although it decreased the synthesis of glucocorticoid receptor mRNA. In addition, ursolic acid did not decrease the expression of c-jun and DNA-binding activity of AP-1 to its cognate sequences. Taken together, we suggest that ursolic acid may induce the repression of matrix metalloproteinase-9 by stimulating the muclear translocation of glucocorticoid receptor, and the translocated glucocorticoid receptor probably down-modulating the trans-activating function of AP-1 to 2-0-teradecanoylphorbol-acetate responsible element of matrix metalloproteinase-9 promoter region.

Expression of matrix metalloproteinase-2 and survivin in endometrioid and nonendometrioid endometrial cancers and clinicopathologic significance

Evren Yilmaz,Meral Koyuncuoglu,İlknur Bilkay Görken,Emre Okyay,Bahadir Saatli,Emine Cagnur Ulukus,Ugur Saygili 대한부인종양학회 2011 Journal of Gynecologic Oncology Vol.22 No.2

Objective: To determine matrix metalloproteinase-2 and survivin expressions in endometrial cancers, their relation to clinical and histologic parameters and to investigate any difference in the expression of these markers between endometrioid and nonendometrioid cancers. Methods: Ninety-five patients with endometrial cancer, were included. Matrix metalloproteinase-2 and survivin expressions were analyzed immunohistochemically from paraffin-embedded tissues by using specific monoclonal antibodies. Results: Survivin nuclear expression was higher in endometrioid cancer as compared to nonendometrioid cancer (p=0.040), but there was no difference for cytoplasmic survivin and matrix metalloproteinase-2 expressions between type I and type II carcinomas. Survivin cytoplasmic staining was significantly lower in patients with deep myometrial invasion (p=0.038). Nuclear expression of survivin is decreased in histologic grade 3 tumors compared to grade 1 and 2 tumors (p=0.013), but there is no difference between grade 1 and 2. We did not find any statistically significant difference between survivin or matrix metalloproteinase-2 expressions and survival. Conclusion: Survivin and matrix metalloproteinase-2 are present in endometrioid and nonendometrioid cancers. Grade 1 and 2 tumors and carcinomas having myometrial invasion less than 50% have higher survivin expression. These results supports that, survivin may play an important role in early stage tumors and early phases of tumor development. We did not find any association between matrix metalloproteinase-2 expression and classical prognostic factors in endometrial cancer and both proteins were not associated with survival. Objective: To determine matrix metalloproteinase-2 and survivin expressions in endometrial cancers, their relation to clinical and histologic parameters and to investigate any difference in the expression of these markers between endometrioid and nonendometrioid cancers. Methods: Ninety-five patients with endometrial cancer, were included. Matrix metalloproteinase-2 and survivin expressions were analyzed immunohistochemically from paraffin-embedded tissues by using specific monoclonal antibodies. Results: Survivin nuclear expression was higher in endometrioid cancer as compared to nonendometrioid cancer (p=0.040), but there was no difference for cytoplasmic survivin and matrix metalloproteinase-2 expressions between type I and type II carcinomas. Survivin cytoplasmic staining was significantly lower in patients with deep myometrial invasion (p=0.038). Nuclear expression of survivin is decreased in histologic grade 3 tumors compared to grade 1 and 2 tumors (p=0.013), but there is no difference between grade 1 and 2. We did not find any statistically significant difference between survivin or matrix metalloproteinase-2 expressions and survival. Conclusion: Survivin and matrix metalloproteinase-2 are present in endometrioid and nonendometrioid cancers. Grade 1 and 2 tumors and carcinomas having myometrial invasion less than 50% have higher survivin expression. These results supports that, survivin may play an important role in early stage tumors and early phases of tumor development. We did not find any association between matrix metalloproteinase-2 expression and classical prognostic factors in endometrial cancer and both proteins were not associated with survival.

성견에서 하악골 신장술 후 하악과두 연골의 조직학적 변화와 Matrix Metalloproteinase-2 (MMP-2)와 Tissue Inhibitor of Matrix Metalloproteinase-2 (TIMP-2)의 발현

변준호,박봉욱,조영철,성일용,손재희,김종렬,Byun, June-Ho,Park, Bong-Wook,Cho, Yeong-Cheol,Sung, Iel-Yong,Son, Jae-Hee,Kim, Jong-Ryoul 대한악안면성형재건외과학회 2006 Maxillofacial Plastic Reconstructive Surgery Vol.28 No.5

Purpose : This study was to clarify the changes in mandibular condyle after unilateral mandibular distraction osteogenesis throughout histological changes and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2). Materials & Methods : Intraoral distractors were placed via submandibular incision in 8 dogs. Two unoperated animals served as controls. Distraction was performed five days after osteotomy as a rate of 0.5 mm twice per day for 10 days. Two animals were sacrificed on 7, 14, 28, and 56 days after completion of distraction, respectively. Ipsilateral condyles were harvested and processed for histological and immunohistochemical examinations. Results : The condyle cartilage is separated into four layers: fibrous layer, proliferative layer, hypertrophic layer, and calcified layer. At 7 days and 14 days after distraction, the condylar cartilage showed the decreased thickness of the articular cartilage and reduced cellularity. At 28 days after distraction, there was an increase in cellularity of fibrous, proliferative, and hypertrophic layer. However, it demonstrated reduced cellularity compared to the control. At 56 days of after distraction, the articular cartilage was an almost normal histologic structure. Positive Safranin-O staining, indicative of sulfated proteoglycans, was examined in the condylar cartilge of nonloaded control. At 7 days and 14 days after distraction, the sulfated proteoglycans is almost completely depleted from the noncalcified part of the condylar cartilage. At 28 days after distraction, there was an increase in Safranin-O staining intensity. However, the staining intensity of the experimental condyle was weaker than that of the control. At 56 days of after distraction, the condylar cartilage showed almost normal Safranin-O staining pattern. In control condyle, MMP-2 immunostaining was seen in fibrous, proliferative, and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in fibrous and proliferative layer. At 7 days and 14 days after distraction, strong MMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, MMP-2 immunostaining was seen in the fibrous and hypertrophic layer of condylar cartilage, however, their immunoactivity was reduced. At 56 days after distraction, MMP-2 immunoreactivity showed almost normal immunostaining pattern. In control condyle, TIMP-2 immunostaining was primarily seen in fibrous and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in proliferative layer. At 7 days after distraction, very weak TIMP-2 immunoreactivity appeared in fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 14 days after distraction, weak TIMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, TIMP-2 immunoreactivity was increased in the fibrous and hypertrophic layer of condylar cartilage. At 56 days after completion of distraction, TIMP-2 immunoreactivity showed almost normal immunostaining pattern. Conclusions : The results show that short-term outcome of physiologic distraction osteogenesis may lead to degenerative changes in the condylar cartilage. These alterations in the condylar cartilage may be considered as a pressure-related degeneration of the cartilage tissue. However, the long-term results suggest that the condylar cartilage display repair activity after mandibular distraction osteogenesis.

다양한 표피 종양에서 Matrix Metalloproteinase-2와 Matrix Metalloproteinase-9 발현에 관한 면역조직화학적 연구

박홍표,최규철,정병수 조선대학교 부설 의학연구소 2002 The Medical Journal of Chosun University Vol.27 No.1

Background and Objectives: Degradation of basement membranes and extracelluar matrix is a essential step in tumor cell migration, invasion and metastasis formation. Two types of Matrix metalloproteinases (MMPs), MMP-2 and MMP-9 are active in the digestion of type Ⅳ collagen, the main constituent of the basement membrane. Based on this background, we investigated the expression patterns of Type Ⅳ collagenase (MMP-2 and MMP-9) in squamous cell carcinoma, adenoid (acantholytic) squamous cell carcimoma, and its precancerous conditions, actinic keratosis and Bowen's disease by immunohistochemical technique. Materials and Methods: Formalin-fixed and paraffin-embedded tissues from 5 actinic keratosis, 5 Bowen's diseases, 5 adenoid squamous cell carcinoma, and 5 squamous cell carcinoma were immunolabelled with monoclonal antibodies directed against MMP-2 and MMP-9. Result: For MMP-2, actinic keratosis, Bowen's disease, adenoid squamous cell carcinoma, and squamous cell carcinoma was expressed to relative low level in tumor cells. Bowen's disease showed a negative staining for MMP-9, but in microinvasive area of actinic keratosis and adenoid lining cells of adenoid squamous cell carcinoma was a positive staining or MMP-9. In Bowen's disease, squamous cell carcinoma, and adenoid squamous cell carcinoma, dyskeratotic cells and acantholytic cells were stained for MMP-2 but for MMP-9, only in adenoid squamous cell carcinoma. Conclusion: This finding suggested that MMP-9 may be related to tumor invasion to surrounding stroma and that MMP-2 and MMP-9 may be involves in the apoptotic process of the tumor cells.

방사선조사를 받은 흰쥐 소장 점막의 손상과 재생과정 중 금속단백효소 및 억제자의 발현

곽현주,이경자,이정식 대한방사선종양학회 2003 Radiation Oncology Journal Vol.21 No.1

Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the RatHyon Joo Kwag, M.D.*, Kyoung Ja Lee, M.D.†, and Chung Sik Rhee, M.D.‡

Matrix Metalloproteinase: Inhibitory Effect of Marine Substances on MMP-2 and MMP-9

Nguyen, Van-Tinh,Qian, Zhong-Ji,Jung, Won-Kyo The Basic Science Institute Chosun University 2011 조선자연과학논문집 Vol.4 No.4

Marine ecosystems are often characterized by a high biological diversity, and it corresponds to a high chemical diversity. Up to present, more than 20,000 new bioactive substances have been isolated from marine organisms, where considerable numbers of these naturally occurring derivatives are developed as potential candidates for pharmaceutical application. In this process, screening of natural products from marine organisms that could potentially inhibit the expression of metalloproteinases has gained a huge popularity. Cancer is considered as one of the deadliest diseases in the medical field. Matrix metalloproteinase (MMPs) can degrade extracellular matrix (ECM) components and play important roles in a variety of biological and pathological processes. Matrix metalloproteinase inhibitors (MMPIs) have been identified as potential therapeutic candidates for metastasis, arthritis, chronic inflammation and wrinkle formation.

토끼에서 Myocardial Infarction 후 Left Ventricular Remodeling에 대한 Matrix Metalloproteinase의 차단 효과

김수현,한승세,정태은,홍그루 대한흉부외과학회 2007 Journal of Chest Surgery (J Chest Surg) Vol.40 No.5

배경: Matrix metalloproteinase (MMP) 차단은 심근경색 후 좌심실 확장에 대한 가능한 치료 전략으로 대두되고 있다. 선택적 MMP 차단제의 투여가 심근경색 후 초기 단계에 MMP가 대량으로 분비되는 짧은 기간을 차단하는 것이 좋을 것인지, 초기 전체 기간 동안 차단하여야 할 것인지를 알아보고자 하였다. 대상 및 방법: 토끼를 이용하여 기관 삽관 하에 전신 마취를 하고 흉골 정중절개한 다음 좌전하행지 관상동맥을 결찰하여 심근경색을 만들었다. 실험군은 3군으로 나누었다. 심근경색 단독(MI only 군)군은 7예, MMP 차단제 5일 투여군(MMPI 5d 군)은 6예, MMP 차단제 9일 투여군(MMPI 9d 군)은 5예이었다. MMP 차단제로는 MMP-2와 MMP-9에 대한 선택적 차단제인 CG2300을 사용하였다. 각 군은 심장초음파도 검사를 4회 시행하였는데, 술 전, 술 후 1주, 2주 및 3주에 하였다. 검사는 2D 심초음파도를 사용하여 EDD, ESD 및 EF를 측정하였다. 술 후 4주에 희생한 토끼의 심장을 western blotting과 zymography를 하여 MMP-2와 MMP-9의 단백질과 활성의 변화를 조사하였고, 경색부위를 병리학적 조직검사를 하였다. 결과: 심초음파도 검사상, MI only군에서는 대체로 술 전에 비하여 술 후 EDD와 ESD가 증가한 추세로 좌심실이 확장하였음을 알 수 있었다. MMP 차단제 9일 투여군에서는 심근경색 단독군과 MMP 차단제 5일 투여군에 비해 좌심실의 확장이 감소한 경향을 보였다. EF 는 MMP 차단제 9일 투여군에서 술 후에 술 전과 큰 변동이 없었으며, 다른 군들보다 높은 경향이었다. MMP 단백질의 발현과 활성 변화를 보면, 심근경색 단독군, MMP 차단제 5일 및 9일 투여군 등 3군을 정상 심장군과 비교하였을 때 MMP-2 단백질 발현과 활성 변화는 일어나지 않았다. 그리고 MMP-9의 단백 발현 및 활성은 검출되지 않았다. 병리학적 조직 소견을 보면 심근경색 단독군에서 심한 교원질 침착이 있었다. MMP 차단제 5일 투여군과 9일 투여군에서는 교원질 축적이 감소된 경향을 보였다. MMP 9일 투여군에서는 모세혈관의 수가 증가한 것을 볼 수 있었다. 결론: 관상동맥을 결찰하여 심근경색을 유도하면 술 후 빠른 시간 내에 심실이 확장되며 MMP 차단제를 투여할 경우 심실의 확장이 완화됨을 알 수 있었다. MMP 차단제의 효과는 초기의 대부분 기간을 차단하는 것이 좋다고 생각된다. MMP 차단제가 혈관신생을 증가시켜 심실 재형성을 완화할 수 있는 것으로 분석된다.

오미자 유산균 발효물의 주름개선 효과

이정희 ( Jung Hee Lee ),김종임 ( Jong Im Kim ),최화정 ( Hwa Jung Choi ),이정현 ( Jung Hyun Lee ) 대한화장품학회 2014 대한화장품학회지 Vol.40 No.4

새로운 주름개선제 성분을 찾기 위해서 본 연구에서는 사람 피부 섬유아세포의 세포독성, 콜라겐 생합성, matrix metalloproteinase-I (MMP-1) 및 elastase 저해활성에 대한 Lactobacillus plantarum으로 발효된 오미자 발효액의 주름개선 효과를 평가하였다. 먼저, 오미자 추출물은 L. rhamnosus으로 37 ℃에서 1일 동안 발효하였다. 발효물의 세포독성은 cytopathic effect reduction 방법에 의해 평가하였다. 콜라겐 생합성에 대한 발효물의 영향은 procollagen type-IC peptide EIA kit에 의해 평가하였으며, matrix metalloproteinase-I (MMP-1)에 대한 발효물의 영향은 Matrix Metalloproteinase-1 Biotrack activity Assay Kit에 의해 평가하였다. Elastase inhibition assay는 기질로써 N-Suc-(Ala)3-nitroanilide을 사용하여 기질 반응에 의해 평가하였다. 결과로써 오미자 발효물은 사람 피부 섬유아 세포에 대해 100 μg/mL의 농도에서 세포독성을 나타내지 않았다. 또한 오미자 발효물은 콜라겐 생합성을 촉진시켰으며, MMP-1의 저해 효과를 나타내었다(p <0.05). Elastase inhibition assay에서 오미자 발효물의 IC50은 36.4 μg/mL이었다. 그러므로 본 연구에서 오미자 발효물은 주름개선 효과를 보유하고 있으며, 이것은 피부의 주름개선을 위해 사용가능하리라 사료된다. To identify new active anti-wrinkle ingredients, this study investigated the anti-wrinkle effects of Schizandra chinensis Baillon fermented with Lactobacillus plantarum (SCF) by assessment of cytotoxicity of human dermal fibroblast, collagen biosynthesis, matrix metalloproteinase-I (MMP-1) inhibition and elastase inhibition. S. chinensis was fermented with L. rhamnosus for 1 day at 37 ℃. The cytotoxicity of SCF was evaluated by a cytopathic effect reduction method. Effects on collagen biosynthesis and matrix metalloproteinase-I (MMP-1) of SCF were evaluated by previous reported method using procollagen type-IC peptide EIA kit and Matrix Metalloproteinase-1 Biotrack activity Assay Kit, respectively. Elastase inhibition assay was conducted by reaction of enzyme and substrate using N-Suc-(Ala)3-nitroanilide as the substrate. As the results, SCF didn’t show cytotoxicity against human dermal fibroblast at concentration of 100 μg/mL. Also, SCF was increased collagen synthesis and showed inhibitory effect of MMP-1 (p < 0.05). In the elastase inhibition assay, the IC50 of SCF was 36.4 μg/mL. Therefore, our results indicated that SCF possesses anti-wrinkle effects and can be used practically for anti-wrinkle care of skin.

포스터 발표 : 담관내 성장형과 비담관내 성장형 담관상피암종에서 matrix metalloproteinase-2 와 -9 및 tissue inhibitor of metalloproteinase-1 과 2 발현

채광조,라선영,오봉경,구자승,김영주,박찬일,박영년 대한간학회 2003 Clinical and Molecular Hepatology(대한간학회지) Vol.9 No.3(S)

Background/Aims: The intraductal growth (IG) type of cholangiocarcinoma has a better prognosis than the mass forming (MF) and periductal infiltrating (PI) types. Matrix metalloproteinase (MMP)-2 and MMP-9, regulated by tissue inhibitor of metalloproteinases (TIMP)-2 and TIMP-1, respectively, play important roles in the degradation of the basement membrane during tumor invasion. Alteration in MMPs and TIMPs may regulate the gross morphology of cholangiocarcinoma. Methods: MMP-2 and MMP-9 were analyzed in 22 cholangiocarcinomas with fresh tissue using zymography and real time quantitative RT-PCR, and expressions of MMP-2, MMP-9, TIMP-1 and TIMP-2 were evaluated in 76 cholangiocarcinomas by immunohistochemistry. Results: The total MMP-2, active 62 kDa MMP-2, and MMP-2 mRNA increased 5.6 and 8.0 times, 5.3 and 7.9 times, and 3.1 and 6.8 times in the MF and PI types, respectively, compared to the IG type. The MMP-9 was present in the proform without activation and revealed no significant difference in relation to the gross types. The balanced expressions of MMP-2/TIMP-2 and MMP-9/TIMP-1 were significantly decreased in the MF and PI types, compared to the IG type. In the non-neoplastic liver, the expressions of MMP-2 and MMP-9 were very low without active form. Conclusions: The activities of MMP-2 and loss of balanced expressions of MMP-2/TIMP-2 and MMP-9/TIMP-1 were increased in the MF and PI types with invasive growth compared to the IG type with a preserved basement membrane. MMP-2 and imbalance between MMPs and TIMPs are suggested to play important roles in invasive growth related to the gross type of cholangiocarcinoma

추간 판 조직에서 거대 세포 침윤과 Matrix metalloproteinases의 발현

전창훈,하승연,김한겸 대한척추외과학회 2002 대한척추외과학회지 Vol.9 No.1