MMT and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide

Yang, Won-Kyung,Lee, Woo Cheol,Kim, Mi-Ri,Son, Ho-Hyun 大韓齒科保存學會 2005 Restorative Dentistry & Endodontics Vol.30 No.5

이 연구에서는 Prevotella nigrescens (P. nigrescens)의 lipoplysaccharide (LPS)로 자극한 치주인대 섬유아세포에서 matrix metalloproteinase (MMP)와 tissue inhibitor of metalloproteinase (TIMP)의 생성 양상과, LPS를 수산화칼슘으로 처리했을 때의 영향을 평가하였다. P. nigrescens에서 추출, 정제한 여러 농도의 LPS의 수산화칼슘으로 처리한 LPS로 치주인대 섬유아세포를 자극하여, Immunoprecipitation법으로 MMP-1, -2, TIMP-1의 단백질 생성 양상을, real-time polymerase chain reaction법으로 MMP-1의 mRNA 발현 양상을 분석하였다. 이 연구의 결과는 아래와 같다. 1. MMP-1은 단백질과 유전자 수준 모두 자극 시간과 비례하여 증가하여 48시간에 최대값을 보였다. 2. MMP-2 단백질 생성은 1, 10㎎/㎖에서 자극 시간과 비례하여 증가하였다. 3. TIMP-1 단백질 생성은 24시간까지 증가하다가 48시간에 감소하였고, 0.1과 1 ㎍/㎖에서 증가하였으나 10㎍/㎖에서 억제되었다. 4. P.nigrescens의 LPS를 수산화칼슘으로 처리시 MMP-1의 mRNA 발현은 현저하게 감소하였다. The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periedontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purifled from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ㎍/㎖) or LPS (10 ㎍/㎖) preetreated with 12.5 ㎎/㎖ of Ca (OH)_(2) for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were pelformed for protein level analysis of MMP-1, MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitativre polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The production of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner. and showed maximum value at 48 h in both prootein and mRNA level. But there was no dose-depen-dent increase. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ㎍/㎖ LPS, but there was no dose-dependent increase. 3. TIMP-1 production increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1㎍/㎖ LPS, but suppressed at 10 ㎍/㎖. 4. P. nigrescens LPS pretreated with Ca (OH)_(2) markedly downregulated MMP-1 gene expression.

Type I Collagen-induced Pro-MMP-2 Activation is Differentially Regulated by H-Ras and N-Ras in Human Breast Epithelial Cells

Kim, In-Young,Jeong, Seo-Jin,Kim, Eun-Sook,Kim, Seung-Hee,Moon, A-Ree Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.5

Tumor cell invasion and metastasis are often associated with matrix metalloproteinases (MMPs), among which MMP-2 and MMP-9 are of central importance. We previously showed that H-Ras, but not N-Ras, induced invasion of MCF10A human breast epithelial cells in which the enhanced expression of MMP-2 was involved. MMP-2 is produced as a latent pro-MMP-2 (72 kDa) to be activated resulting the 62 kDa active MMP-2. The present study investigated if H-Ras and/or N-Ras induces pro-MMP-2 activation of MCF10A cells when cultured in two-dimensional gel of type I collagen. Type I collagen induced activation of pro-MMP-2 only in H-Ras MCF10A cells but not in N-Ras MCF10A cells. Induction of active MMP-2 by type I collagen was suppressed by blocking integrin ${\alpha}2$, indicating the involvement of integrin signaling in pro-MMP-2 activation. Membrane-type (MT)1-MMP and tissue inhibitor of metalloproteinase (TIMP)-2 were up-regulated by H-Ras but not by N-Ras in the type I collagen-coated gel, suggesting that H-Ras-specific up-regulation of MT1-MMP and TIMP-2 may lead to the activation of pro-MMP-2. Since acquisition of pro-MMP-2 activation can be associated with increased malignant progression, these results may help understanding the mechanisms for the cell surface matrix-degrading potential which will be crucial to the prognosis and therapy of breast cancer metastasis.

MMP-2 and MMP-9 Inhibitory Effects of Different Solvent Fractions from Corydalis heterocarpa

Ga Hyun Yu(유가현),Fatih Karadeniz(카라데니즈 파티),Chang-Suk Kong(공창숙) 한국생명과학회 2021 생명과학회지 Vol.31 No.11

염주괴불주머니(Corydalis heterocarpa)는 두해살이 풀로 노란 꽃이 피고 염주처럼 잘록한 열매를 맺는 것이 특징이다. 또한 우리나라 갯벌에서 서식하는 염생식물로 내건성, 내염성과 같은 독특한 생리적 기전을 가지고 있으며 민간에서 진통 경련 완화 치료제로 사용하고 있다. 본 연구에서는 항산화, 항염증, 항노화 등의 효과가 있는 것으로 보고되어 있는 염주괴불주머니(Corydalis heterocarpa)을 이용하여 용매분획물(H₂O, n-BuOH, 85% aq. MeOH, n-hexane)을 제조하였으며 이를 이용하여 PMA로 유도된 인간 섬유육종세포 HT-1080 세포에서 MMP-2, MMP-9의 발현 조절에 미치는 영향을 확인하였다. 염주괴불주머니 분획물 처리시 TIMP-1 및 TIMP-2를 증가시키면서 MMP-2 및 MMP-9의 mRNA 및 단백질 발현을 감소시키는 것으로 나타났다. 또한 염주괴불주머니 용매분획물 처리시 MAPKs 신호 전달 경로인 p38, JNK, ERK의 인산화를 억제하여 MMPs 발현을 감소시키는 것을 확인할 수 있었다. 특히 염주괴불주머니의 85% aq. MeOH와 n-hexane 용매분획물에서 MMP-2 및 MMP-9 발현이 효과적으로 억제되어 MMP 억제활성이 높은 물질이 들어있을 것으로 사료되며, 이를 통해 염주괴불주머니의 암 전이 억제 소재 개발을 위한 기초자료로 활용될 수 있을 것으로 기대된다. Natural products have always been an attractive source in terms of novel anti-metastatic compounds which can hinder MMP expression and activity. Corydalis heterocarpa is a salt marsh plant found in the seashores throughout Korea. Its yellow flowers and spikes have been an ingredient in folk medicine to treat spasm and contractions. The present study assessed the potential of different solvent-based fractions from the crude extract of Corydalis heterocarpa (CHE), a halophyte with reported bioactivities, to suppress the PMA-induced MMP expression in human fibrosarcoma HT-1080 cells. The solvent fractions which were named after the solvent used for fractionation (n-hexane, 85% aqueous (aq.) methanol (MeOH), n-butanol (BuOH), and H₂O were shown to inhibit the both elevated mRNA and protein expression levels of MMP-2 and MMP-9 and simultaneously relieved the suppression on the expression of the endogenous MMP inhibitors TIMP-1 and TIMP-2. Results indicated that the CHE fractions might intervene with the PMA-induced activation of the MAPK signaling which is the upstream activator of MMP overexpression. Among tested samples, 85% aq. MeOH and n-hexane fractions of CHE was determined to be the most active and future studies to isolate the bioactive substances responsible for the regulation of the MMP expression are, therefore, urged. In conclusion, C. heterocarpa was shown to be a potential source of anti-metastatic compounds and n-Hexane and MeOH fractions might yield lead molecules to develop novel MMP inhibitors.

MMP-9로 자른 피브로넥틴에 의한 뇌종양세포 U87MG Cells와 단핵구 U937 Cells간의 부착증가 효과

김연향 한국산업기술융합학회 2024 산업기술연구논문지 (JITR) Vol.29 No.2

본 논문에서는 MMP-9에 의해 분해된 피브로넥틴이 종양세포에 대한 면역세포 단핵구의 부착을 촉진하는 주요역할을 한다는 것을 규명한 것이다. MMP-9가 종양세포의 세포외 기질 피브로넥틴에 변화를 일으키고 단핵구의 부착을 증가시키는지 여부를 확인하기 위해, MMP-9를 발현하는 인간 뇌종양 세포인 U87MG 세포주를 transfection 방법으로 구축하고 MMP-9 발현을 면역블로팅과 자이모그램방법으로 확인하였다. MMP-9를 발현하는 U87MG 세포에는 인간 단핵구 U937 세포가 유의하게 부착되었지만, MMP-9를 발현하지 않는 U87MG 세포에는 부착되지 않았다. MMP-9에 의해 분해된 피브로넥틴이 단핵구 U937세포의 부착을 증가시키는 역할을 직접 입증하기 위해, MMP-9로 처리된 피브로넥틴에 단핵구 U937 세포가 부착함을 확인하였다. 이러한 결과는 MMP-9에 의해 잘려진 피브로넥틴이 면역세포와 종양세포 사이의 부착을 중재하는 데 중요한 역할을 할 수 있다는 것을 보여준 것이다. This study investigated the pivotal role of fibronectin cleaved by MMP-9 in enhancing the attachment of monocytes to tumor cells. To determine whether MMP-9 induces changes in the extracellular matrix fibronectin of tumor cells and increases monocyte adhesion, human glioblastoma U87MG cells expressing MMP-9 were established by transfection. The human monocyte U937 cells adhered significantly to U87MG cells expressing MMP-9, however, they did not adhere to U87MG cells that did not express MMP-9. To directly demonstrate the role of MMP-9-cleaved fibronectin in increasing monocyte U937 cell adhesion, the attachment of monocyte U937 cells to fibronectin treated with MMP-9 was confirmed. These findings suggest that MMP-9 alters the extracellular matrix, specifically fibronectin, and plays a crucial role in mediating the adhesion between immune and tumor cells.

Prediction of recurrence in locally advanced squamous cell carcinoma of the cervix by expression of MMP-2 and MT1-MMP

( Nan Hee Jeong ),( Hye Min Yeo ),( Jae Won Kim ),( Kyu Wan Lee ),( Young Sik Kim ),( Nak Woo Lee ) 대한산부인과학회 2008 Journal of Womens Medicine Vol.1 No.1

Objective: The aim of the study was to evaluate the role of matrix metalloproteinase (MMP)-2, tissue inhibitor of metalloproteinase (TIMP)-2, and membrane type metalloproteinase (MT1-MMP) as predictors of recurrence in patients with locally advanced squamous cell carcinoma of the cervix. Methods: We analyzed 48 samples of uterine cervical squamous cell carcinoma (31 cases without recurrence and 17 cases with recurrence) and 20 control samples for the presence of MMP-2, TIMP-2 and MT1-MMP using in situ hybridization. Expression of MMP-2 protein was evaluated using immunohistochemistry. Results: The staining results for MMP-2 and MT1-MMP in patients with recurrence were significantly higher than in cases without recurrence (p=0.006 for MMP-2; p=0.024 for MT1-MMP). The control samples had no intense signal identified for MMP-2, TIMP-2, and MT1-MMP. Tumor size in patients with recurrence was larger than the tumors in cases without recurrence (mean 3.83 cm vs. 1.3 cm; p<0.05). Conclusions: The results of this study suggest that the presence of an intense signal for MMP-2 and MT1-MMP in tumor cells may be used to predict recurrence in stages Ib and IIa squamous cell carcinoma of the cervix.

The comparison of inflammatory mediator expression in gingival tissues from human chronic periodontitis patients with and without type 2 diabetes mellitus

주상돈,이재목,Joo, Sang-Don,Lee, Jae-Mok The Korean Academy of Periodontoloy 2007 Journal of Periodontal & Implant Science Vol.37 No.2

치주질환의 병원균은 세포벽의 항원에 의하여 조직내 존재하는 mononuclear phagocytes가 활성화되어 cytokine들이 생성됨으로써 치주 결체조직의 파괴를 진행시킨다. 이런 관련된 cytokine들은 순차적으로 상주하는 치은세포 및 대식세포가 Matrix metalloproteinase 합성을 하도록 유도하여 조직파괴를 시작한다. 당뇨병은 치주질환의 위험요소중 하나로 당뇨 환자에서는 치주질환의 유병율이 일반인에 비해 높고 치주질환의 중증도도 더 심하여 진행도 빠르다고 알려져 있다. 그 병리 기전 중 하나로 당뇨 환자에서는 치은 열구액 내 중성구 유래의 Matrix metalloproteinase의 활성 증가 및 TIMP-1 발현의 변화가 추정되고 있으나 아직 치은조직내에서 이에 대한 연구는 미흡한 실정으로 이에, 본 실험에서는 제2형 당뇨병 환자와 비당뇨 환자들에서 만성 치주염 부위의 치은 및 건강한 치은에서 염증 매개체 중 하나인 $IL-1{\beta}$, MMP-13 및 TIMP-1의 발현에 대해 상호 비교 분석함으로서 염증, 혈당이 미치는 영향을 밝히고 제 2형 당뇨병 환자에서 심한 치주조직 파괴의 기전을 연구하고자 하였다. 경북대학교병원 치주과 내원환자 중 제2형 당뇨병 환자와 비당뇨 환자들 및 치주질환이 없는 건강인 대조군을 대상으로 여러 가지 환자요소, 임상 치주상태를 기록하고, 전신적으로 건강한 환자의 건강한 부위(n=8, Group 1), 전신적으로 건강한 환자의 만성 치주염 부위(n=8, Group 2), 제2형 당뇨병 환자의 만성 치주염 부위 (n=8, Group 3)에서 각각 변연치은을 채득하고 액화질소에 급속 동결하였다. Western blotting을 이용하여 각 조직 내 $IL-1{\beta}$, MMP-13 및 TIMP-1의 발현을 관찰, densitometer를 이용하여 상대적 발현을 정량, 각 조직의 ${\beta}-actin$을 이용하여 표준화하여 실험군과 대조군들의 평균치를 비교하였다. 비당뇨 환자들의 만성 치주염 부위 및 제2형 당뇨병 환자의 만성 치주염 부위에서 모두 건강 대조군에 비해, MMP-13와 TIMP-1의 발현이 증가되었으며, 또한 $IL-1{\beta}$와 MMP-13는 2형 당뇨 환자의 만성 치주염 부위가 비당뇨 환자의 만성 치주염 부위보다 증가된 발현양상을 보였다. 2형 당뇨 환자의 만성 치주염 부위에서 $IL-1{\beta}$가 증가함에 따라 MMP-13이 증가하였으며, $IL-1{\beta}$ 및 MMP-13이 증가하였지만 TIMP-1 수치는 비당뇨 환자의 만성 치주염환자와 유사한 양상을 보였다. 결론적으로 MMP-13과 TIMP-1이 염증조직에서 증가하는 양상을 보였으며 $IL-1{\beta}$, MMP-13이 2형 당뇨환자의 만성치주염 진행과정에 부분적인 기여 인자로써 역할을 하는 것으로 생각된다. Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was divided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflamed gingiva from patients with chronic periodontitis. Group 3 (n=8) is inflamed gingiva from patients with chronic periodontitis associated with type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantification of $IL-1{\beta}$, MMP-13 and TIMP-1 were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. 1. The expressions of MMP-13 and TIMP-1 showed increasing tendency in group 2 & 3 compared to group 1. 2. The expressions of $IL-1{\beta}$ & MMP-13 were showed increasing tendency in group 3 compared to group 2. 3. As $IL-1{\beta}$ levels were increasing, MMP-13 showed increasing tendency in group 3, and although $IL-1{\beta}$ , MMP-13 levels were increasing, TIMP-1 levels were similar expressed comparing to group 2. In conclusion, this study demonstrated that the expression levels of MMP-13 and TIMP-1 had increasing tendency in inflamed tissue. It can be assumed that $IL-1{\beta}$ and MMP-13 may be partly involved in the progression of periodontal inflammation associated to type 2 DM.

알러젠 제거 옻나무 추출물이 종양 전이 억제에 미치는 영향

박재현,문구,Park, Jae-Hyun,Moon, Goo 대한암한의학회 2010 大韓癌韓醫學會誌 Vol.15 No.1

Objective : The aim of this present study is to evaluate the inhibitory effect of allergen removed Rhus verniciflua (ARV) on Matrix Metalloproteinase-9 (MMP-9), Matrix Metalloproteinase-2 (MMP-2) which is considered to have a clinically important role in tumor metastasis. Methods : The inhibitory effects of standardized extract of ARV on the MMP-2, MMP-9 were investigated by spectrofluorometer while the inhibitory effects on the active MMP-2, pro MMP-2, pro MMP-9 were investigated by zymography. Antimetastatic effect of standardized extract of ARV was investigated in vitro on human fibrosarcoma cell (HT1080)'s invasion through Matrigel. Results : The standardized extract of ARV showed inhibitory effects on the active MMP-2 (IC50, $1.01{\mu}g$/ml), active MMP-9 (IC50, $2.5{\mu}g$/ml) depending on concentrations which was determined by spectrofluorometer. The standardized extract of ARV showed inhibitory effects on the active MMP-2, pro MMP-2, pro MMP-9 depending on concentrations which was determined by zymography. However its inhibitory effect on pro MMP-9 was relatively weaker rather than active MMP-2, pro MMP-2. The standardized extract of ARV showed inhibitory effects in vitro on human fibrosarcoma cell (HT1080)'s invasion through Matrigel according to concentration. Conclusions : These results indicate that standardized extract of ARV has antimetastatic effect through inhibit again MMP-2, MMP-9. Also its inhibitory effect is more powerful on active MMP-2, pro MMP-2 than on active MMP-9, pro MMP-9. It is necessary to conduct further studies on other MMP families, TIMP, and each component of standardized extract of ARV.

한국의 준연동형 비례대표 선거제도개혁과 집합적 전략투표 동원

강우진 서강대학교 현대정치연구소 2020 현대정치연구 Vol.13 No.2

This study analyzes the parties’ mobilizing strategy of the collective strategic voting in the previous three cases (Albania, Lesotho, and Venezuela) that have experienced the MMP system. In doing so, this study aims to critically review the electoral reform induced before the 21st National Election. The electoral reform of the quasi-MMP electoral system in Korea was urgently introduced ahead of the 21st National Assembly's election by a party coalition pursuing power maximization. Accordingly, it was done only with the focus of introducing the MMP system itself to increase proportionality without forming consensus among voters. The experience of the three countries analyzed in this article confirmed that the MMP system is vulnerable to parties' mobilization of the collective strategic voting of voters. In the end, in the three cases reviewed by this study, the MMP system failed to maintain its original form at the time of introduction. Albania abandoned MMP and adopted a proportional representation system, and Venezuela converted to MMM. Lesotho maintained the MMP but changed the election system from 2 votes per person to 1 vote per person to prevent strategic misuse. To make the expected effects of MMP electoral reform realize, it is crucial to forming a consensus among voters, the sovereign, to introduce the MMP election system. When voters actively support the introduction of the MMP election system, the possibility of collective strategic voting will significantly reduce. In this case, the political party's political incentive to circumvent the MMP system by creating a satellite political party will decrease. 이 글의 목적은 MMP 제도를 경험했던 앞선 세 사례(알바니아, 레소토, 베네주엘라)에서 나타난 집합적 전략투표의 동원 전략에 초점을 맞추어 한국의 준연동형 선거제도 개혁을 검토 하는 것이다. 한국의 준연동형 선거제도 개혁은 권력극대화를 추구하는 정당연합에 의해서 제21대 국회의원 선거를 앞두고 급박하게 이루어졌다. 이에 따라서 선거제도 개혁에 대한 사 회적 합의를 형성하지 못한 채 비례성의 확대만을 위해서 제도 도입 자체에만 초점을 맞추어 이루어졌다. 세 나라의 경험은 MMP 제도가 유권자들의 집합적인 전략투표를 유도하는 정당 의 동원 전략에 취약하다는 것을 확인해 주었다. 결국 세 사례에서 MMP 제도는 도입당시 원형을 유지하지 못했다. 알바니아는 MMP를 포기하고 비례대표제를 채택하였으며 베네수엘 라는 MMM으로 전환하였다. 레소토는 MMP를 유지하였지만 전략적 오용을 방지하기 위해 서 1인 2표에서 1인 1표로 선거제도를 변경하였다. MMP 선거제도가 기대했던 효과를 만들어내기 위해서는 MMP 제도를 포함한 앞선 선거 제도 개혁의 경험에 대한 엄밀한 논의가 필요하다. 또한, MMP 선거제도에 대한 주권자인 유권자의 공감대 형성이 매우 중요하다.

멜라닌세포성 모반과 악성 흑색종의 MMP-2와 MMP-9 metalloproteinase 발현에 관한 면역조직화학적 연구

신봉석 ( Sin Bong Seog ),박만규 ( Park Man Gyu ),최규철 ( Choe Gyu Cheol ),정병수 ( Jeong Byeong Su ) 대한피부과학회 2003 대한피부과학회지 Vol.41 No.8

N/A Background : One of the most important steps in neoplastic progression is represented by invasion of surrounding normal tissues by neoplastic cells. Enzymes such as the metalloproteinases(MMPs) are thought to be involved in the process of destruction of basement membranes and stromal invasion by tumor cells. Objective : We investigated the expression patterns of MMP-2 and MMP-9 in acquired and congenital melanocytic nevi, and malignant melanoma by immunohistochemical technique. Method : Formalin-fixed and paraffin-embedded tissues from 4 junctional nevi, 4 compound nevi, 5 intradermal nevi, 6 congenital melanocytic nevi, and 6 malignant melanomas were immunolabelled with monoclonal antibodies directed against MMP-2 and MMP-9. Result : The benign melanocytic nevi showed negative or low expression for MMP-2 and MMP-9 with the exception of positive staining in involuting neuroid intradermal nevus, and the expression of MMP-9 was detected in 3 cases of congenital melanocytic nevi. The malignant melanoma exhibited high expression of MMP-2 with variable intensity of reactivity in different areas of the tumors and MMP-9 was found to be focally expressed by the tumor cells in intraepidermal and dermoepidermal junction. These findings suggest that the expression of MMP-2 and MMP-9 may be related to tumorigenesis of melanocytic tumors and MMP-9 may be involved in the early stage of tumor progression. (Korean J Dermatol 2003;41(8) : 991~996)

난소암 세포주들의 전이능력과 Type Ⅳ collagenase 효소 활성화의 비교

이상형(SH Lee),배석년(SN Bae),김재동(JD Kim),박종섭(JS Park),김승조(SJ Kim) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.5

Our purpose in this study was to detect the secretory pattern and the degree of enzymatic activity of MMP-2 and to evaluate the invasive activty in vitro in each cancer cell line, using four kinds of human ovarian cancer cell lines (CAOV-4, SW626, NIH: OVCAR-3, and SK-OV-3) as well as MCF-7MMP-2 cell line which secretes recombinant 72-kDa MMP-2 as an inactive form. The results were as follows; 1. An inactive form of MMP-2 was secreted by all ovarian cancer cell lines, but the activated form of MMP-2 was also secreted by NIH: OVCAR-3, SK-OV-3, and SW626. 2. In the Boyden chamber chemoinvasion assay, the invasive activity was shown in SK-OV-3, NIH: OVCAR-3, SW626, and CAOV-4 cell lines in order. This finding corresponds with the secretory pattern of 62 kDa MMP-2, activated form. 3. In the outgrowth morphology on Matrigel, the dendritic processes of NIH: OVCAR-3 and SK-OV-3 cell lines were seen to emerge more actively than those of SW626 and CAOV-4 cell lines. NIH: OVCAR-3 and SK-OV-3 cell lines also had a clustered formation. 4. An inactive form of MMP-2 secreted by CAOV-4 was activated by the treatment with Concanavalin A, MMP-2 activator. 5. The productions and secretions of MMP-2 in SK-OV-3, NIH: OVCAR-3, and SW626 cell lines were inhibited by the treatment with cycloheximide, MMP-2 inhibitor. From the above results, we showed that the invasion and metastasis of human ovarian cancr cell lines have directly relations with MMP-2. It was identified that the secretory patterns of activated MMP-2 were closely associated with the invasiul activity of human ovarian cancer cell lines in vitro and that the MMP-2 secreted by human ovarian cancer cell lines might been activated or inhibited by enzyme regulators. In the future, we wish this study applied to the metastatic prevention and treatment of human ovarian cancer by using enzyme regulators.