• 생명과학회지
• /
• 제29권3호
• /
• pp.287-294
• /
• 2019

갯메꽃은 해안 사구에 분포하며 식물로 높은 환경 적응력을 가지고 있으며 갯메꽃은 항산화, 해열, 살균, 이뇨작용 등에 효과가 있는 것으로 알려져 있다. 본 연구에서는 암전이 과정에서 중요한 역할을 한다고 알려진 matrix metalloproteinases (MMPs)의 일종인 MMP-2와 MMP-9의 활성에 대한 갯메꽃의 억제효과를 methylene chloride (MC) 및 methanol (MeOH) 추출물과 조추출물, 유기용매 분획물을 시료로 하여 인체 섬유육종 HT-1080 세포를 이용하여 ELISA (Enzyme-linked immunosorbent), gelatin zymography, Wound healing assay, RT-PCR, western blot 실험을 통해 조사하였다. 갯메꽃의 MC 및 MeOH 추출물, 조추출물과 4가지 유기용매 분획물이 HT-1080 세포에서 MMP-2와 MMP-9 생성 억제 활성을 나타냄을 확인하였다. 그 중에서도 n-hexane, 85% aq.MeOH 분획물이 전반적으로 MMP-2와 MMP-9에 대한 높은 억제활성을 보였다. 이들 결과로부터 85% aq.MeOH과 n-hexane 분획물에 MMP 억제 활성이 높은 물질이 존재할 것으로 추측되어지며, 갯메꽃 추출물 및 분획물이 암침윤 및 전이를 억제하는 효과적인 항암소재로서의 가능성이 있음을 제시한다. 이에 본 연구팀은 85% aq.MeOH과 n-hexane 분획물에 있는 활성물질을 분리하기 위한 연구를 추가적으로 수행할 계획이다.

• Nutrition Research and Practice
• /
• 제15권4호
• /
• pp.444-455
• /
• 2021

BACKGROUND/OBJECTIVES: Adipocytes undergo angiogenesis to receive nutrients and oxygen needed for adipocyte' growth and differentiation. No study relating quercetin with angiogenesis in adipocytes exists. Therefore, this study investigated the role of quercetin on adipogenesis in 3T3-L1 cells, acting through matrix metalloproteinases (MMPs). MATERIALS/METHODS: After proliferating preadipocytes into adipocytes, various quercetin concentrations were added to adipocytes, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to evaluate cell proliferation. Glycerol-3-phosphate dehydrogenase (GPDH) activity was investigated as an indicator of fat accumulation. The mRNA expressions of transcription factors related to adipocyte differentiation, CCAAT/enhancer-binding proteins (C/EBPs), peroxisomal proliferatoractivated receptors (PPAR)-γ, and adipocyte protein 2 (aP2), were investigated. The mRNA expressions of proteins related to angiogenesis, vascular endothelial growth factor (VEGF)-α, vascular endothelial growth factor receptor (VEGFR)-2, MMP-2, and MMP-9, were investigated. Enzyme activities and concentrations of MMP-2 and MMP-9 were also measured. RESULTS: Quercetin treatment suppressed fat accumulation and the expressions of adipocyte differentiation-related genes (C/EBPα, C/EBPβ, PPAR-γ, and aP2) in a concentration-dependent manner in 3T3-L1 cells. Quercetin treatments reduced the mRNA expressions of VEGF-α, VEGFR-2, MMP-2, and MMP-9 in 3T3-L1 cells. The activities and concentrations of MMP-2 and MMP-9 were also decreased significantly as the concentration of quercetin increased. CONCLUSIONS: The results confirm that quercetin inhibits adipose tissue differentiation and fat accumulation in 3T3-L1 cells, which could occur through inhibition of the angiogenesis process related to MMPs.

• 대한의생명과학회지
• /
• 제10권2호
• /
• pp.143-151
• /
• 2004

Matrix metalloproteinases (MMPs) are capable of degrading extracellular matrix, a process that is necessary for angiogenesis, tumor invasion and metastasis. Platelet-activating factor (PAP) increases angiogenesis, tumor growth and metastasis through nuclear factor (NF)-$\kappa$B activation. Based on these facts, the involvement of MMPs in PAF-induced pulmonary metastasis was investigated in murine sarcoma cells, MMSV-BALB/3T3. Messenger RNA expression and enzymatic activity of MMP-9 were assessed by RT-PCR and zymography, and cell migration and metastasis were done for the detection of MMP-9 functional activity. PAP induced mRNA expression and enzymatic activity of MMP-9, and its effects were either inhibited by the PAP antagonist, WEB 2170 or by the NF-$\kappa$B inhibitor, parthenolide, or p65 antisense oligonucleotide in a dose-dependent manner. In addition, PAF induced promoter activity of MMP-9, which was inhibited by WEB 2170, phenanthroline, NAC, PDTC. These results indicate that PAF induces mRNA expression and enzymatic activity of MMP-9 in NF-$\kappa$B dependent manner. Cell migration assay showed that PAF induced MMSV-BALB/3T3 migration, and its effect was significantly inhibited by treatment with phenanthroline. PAF enhanced pulmonary metastasis of murine sarcoma cells, MMSV-BALB/3T3 was also reduced by phenanthroline. These results suggest that PAF-enhanced cell migration and pulmonary metastasis is mediated through the expression of MMP. In conclusion, It is suggested that PAF enhances pulmonary metastasis by inducing MMP-9 expression via the activation of NF-$\kappa$B.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권7호
• /
• pp.4107-4113
• /
• 2013

Objective: Matrix metalloproteinase-9(MMP-9) plays an important role in tumor cell invasion. Although it has been studied frequently in ovarian cancer, its prognostic impact is still equivocal. The aim of this study was to more precisely estimate its prognostic significance. Method:We searched Pubmed, Embase, OVID, Sciencedirect and CBM databases to identify eligible studies. Hazard ratios (HRs) or odds ratios (ORs) with 95% confidence intervals (95% CIs) were pooled across studies using fixed-effects or random-effects models. We also performed subgroup analysis. Results: 30 studies (n=2552 patients) focusing on prognosis or expression of MM-9 were included. Increased expression of MMP-9 was associated with poor prognosis in ovarian cancer patients (HR=1.68, 95%CI 1.09-2.59, p=0.02). Besides, MMP-9 expression in ovarian cancer was significantly higher than non-malignant tumors (OR=11.46, 95%CI 8.47-15.50, P<0.00001). Moreover, increased expression of MMP-9 was significantly associated with FIGO stage (OR=4.85, 95%CI 2.60-9.04, P<0.00001), grade of differentiation (OR=3.34, 95%CI 2.46-4.54, P<0.00001), lymph node metastasis (OR=5.75, 95%CI 3.71-8.92, P<0.00001) and there was no association with histological type of ovarian cancer. Conclusions: Increased expression of MMP-9 was associated with poor prognosis in ovarian cancer patients. Down-regulation of MMP-9 is an attractive therapeutic approach which might improve outcome of ovarian cancer.

• Journal of Korean Neurosurgical Society
• /
• 제61권5호
• /
• pp.548-558
• /
• 2018

Objective : Diagnosing acute cerebral infarction is crucial in determining prognosis of stroke patients. Although many serologic tests for prompt diagnosis are available, the clinical application of serologic tests is currently limited. We investigated whether $S100{\beta}$, matrix metalloproteinase-9 (MMP-9), D-dimer, and heat shock protein 70 (HSP70) can be used as biomarkers for acute cerebral infarction. Methods : Focal cerebral ischemia was induced using the modified intraluminal filament technique. Mice were randomly assigned to 30-minute occlusion (n=10), 60-minute occlusion (n=10), or sham (n=5) groups. Four hours later, neurological deficits were evaluated and blood samples were obtained. Infarction volumes were calculated and plasma $S100{\beta}$, MMP-9, D-dimer, and HSP70 levels were measured using enzyme-linked immunosorbent assay. Results : The average infarction volume was $12.32{\pm}2.31mm^3$ and $46.9{\pm}7.43mm^3$ in the 30- and 60-minute groups, respectively. The mean neurological score in the two ischemic groups was $1.6{\pm}0.55$ and $3.2{\pm}0.70$, respectively. $S100{\beta}$, MMP-9, and HSP70 expressions significantly increased after 4 hours of ischemia (p=0.001). Furthermore, $S100{\beta}$ and MMP-9 expressions correlated with infarction volumes (p<0.001) and neurological deficits (p<0.001). There was no significant difference in D-dimer expression between groups (p=0.843). The area under the receiver operating characteristic curve (AUC) showed high sensitivity and specificity for MMP-9, HSP70 (AUC=1), and $S100{\beta}$ (AUC=0.98). Conclusion : $S100{\beta}$, MMP-9, and HSP70 can complement current diagnostic tools to assess cerebral infarction, suggesting their use as potential biomarkers for acute cerebral infarction.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권9호
• /
• pp.4545-4548
• /
• 2012

Background: MMP-3 is a proteolytic enzyme of the matrix metalloproteinase family. Protein degradation which is their fundamental action regulates different activities of tumor cell such as their growth, differentiation, apoptosis, migration, invasion, angiogenesis as well as their resistance to the immune system. Aim: The aim of this study was to determine MMP-3 serum levels in patients with OSCC and investigate if they correlate with clinicopathological features. Method and materials: Using an ELISA kit, we assessed and compared the circulating levels of MMP-3 in blood serum of 45 oral squamous cell carcinoma patients with 45 healthy control samples. Results: The serum MMP-3 level in OSCC patients was significantly higher ($9.45{\pm}4.6$ ng/ml) than healthy controls ($5.9{\pm}3.6$ ng/ml, p<0.001), especially in females and in older patients. However, there was no apparent correlation in serum MMP-3 concentration with the clinico-pathological features such as tumor location, stage, tumor size, nodal status, distant metastasis, histological grade and smoking. Discussion: This result suggests that the measurement of serum MMP-3 concentration might be helpful to diagnose OSCC but not to predict prognosis.

• The Korean Journal of Physiology and Pharmacology
• /
• 제15권4호
• /
• pp.211-216
• /
• 2011

Glioblastoma multiforme is one of the most common and aggressive tumors in central nervous system. It often possesses characteristic necrotic lesions with hemorrhages, which increase the chances of exposure to thrombin. Thrombin has been known as a regulator of MMP-9 expression and cancer cell migration. However, the effects of thrombin on glioma cells have not been clearly understood. In the present study, influences of thrombin on glioma cell migration were examined using Boyden chamber migration assay and thrombin-induced changes in MMP-9 expression were measured using zymography, semi-quantitative RT-PCR, and Western blotting. Furthermore, underlying signaling pathways by which thrombin induces MMP-9 expression were examined. Thrombin-induced migration and MMP-9 expression were significantly potentiated in the presence of wortmannin, a PI3K inhibitor, whereas MAPK inhibitors suppressed thrombin-induced migration and MMP-9 expression in C6 glioma cells. The present data strongly demonstrate that MAPK and PI3K pathways evidently regulate thrombin-induced migration and MMP-9 expression of C6 glioma cells. Therefore, the control of these pathways might be a beneficial therapeutic strategy for treatment of invasive glioblastoma multiforme.

• KSBB Journal
• /
• 제24권5호
• /
• pp.425-431
• /
• 2009

본 연구에서는 U-251-MG 세포를 이용하여 증식, 이동, 침윤 및 단백질분해효소의 분비에 미치는 PSA와 HGF의 효과를 확인하였다. 그 결과, PSA siRNA에 의해 U-251-MG 세포의 증식은 약 37% 억제되었으나, HGF 처리 (10 ng/mL)에 의해 증식이 약 1.4배 증가되었다. PSA siRNA에 의한 U-251-MG 세포의 이동은 약 60%가 억제되었으나, HGF 처리에 의해 이동이 약 1.3배 증가되었다. 또한, PSA siRNA에 의해 U-251-MG 세포의 침윤이 약 67% 억제되었으나 HGF 처리에 의해 세포의 침윤이 약 4.3배 증가되었다. PSA siRNA처리에 의해 MMP-2의 분비는 약 25%, MMP-9의 분비는 약 20% 억제되었으며, HGF에 의해 PSA siRNA에 의해 억제된 MMP-2 및 MMP-9의 분비가 약 2.8배 및 3.5배 증가되었다. HGF 처리에 의해 플라스민의 분비량은 약 14배 증가되었고, PSA를 억제한 U-251-MG 세포에 HGF를 처리한 결과, 플라스민의 분비가 약 1.6배 증가하였다. 또한, PSA siRNA에 의해 MMP-2의 발현은 유의할만한 변화가 없었으나, MMP-9의 발현은 약 85% 억제되었다. PSA를 억제시킨 U-251-MG 세포에 HGF를 처리한 결과, MMP-2의 발현은 약 5.7배, MMP-9의 발현은 약 6.3배 증가되었다. 한편, MMPs의 광범위 억제제인 BB-94 처리에 의해 U-251-MG 세포의 증식, 이동 및 침윤이 유의할만하게 억제 된 것은 MMP-2 및 MMP-9이 U-251-MG 세포의 증식, 이동 및 침윤에 관여할 것임을 시사해준다.

• 생명과학회지
• /
• 제15권2호
• /
• pp.231-235
• /
• 2005

기질 금속단백분해효소(MMP)는 기저막이나 간질성 조직 등에 있는 세포외기질 성분을 분해하여 상처 치유, 태아의 발생, 종양세포의 침윤과 전이 등을 포함하여 조직이 재구성되는 과정에서 생리학적 및 병리학적인 과정 양쪽 모두에 매우 중요한 역할을 한다. 본 연구에서는 MMP-9 유전자의 발현을 억제하는 물질을 천연물에서 탐색하였고, 탐색된 시료 중에서 높은 역가를 가진 낙우송과의 Metasequoia glyptostroboides가 선택되었다. 여러 가지 flavonoid 성분을 가지고 있는 Metasequoia glyptostroboides를 이용하여 4개의 biflavonoid와 2개의 monoflavonoid 구조의 물질을 분리하였고, 이 flavonoid들은 sciadopitysin, isoginkgetin, bilobetin, 2, 3-dihydrohino-kiflavone, luteolin, apigenin으로 정제하여 구조를 밝히고 zymography, WST-1을 이용한 세포독성시험, northern blot 등의 실험을 통하여 각 화합물의 구조적인 특성과 함께 MMP-9 유전자 발현을 억제하는 효과가 있는 것을 확인하였다.

• Journal of Applied Biological Chemistry
• /
• 제61권1호
• /
• pp.9-15
• /
• 2018

우리는 황련에서 분리된 베르베린의 화장품소재로의 활성을 평가하기 위해 항산화와 항주름 효능을 분석하였다. 시료의 독성을 평가하기 위해 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay를 수행하였으며 항산화와 항주름 활성평가를 위해 ROS 생성능, hyaluronic acid 생성능, MMP-2, -9, TIMP-1,-2, $TNF-{\alpha}$의 발현을 각각 측정하였다. 베르베린에 대한 자외선의 세포 독성은 CCD-986sk 섬유아세포를 이용하여 MTT 분석으로 측정되었고, 세포 독성이 $25{\mu}g/mL$의 농도 미만에서 관찰되지 않았다. 또한 베르베린이 ROS 생산을 농도 의존적으로 감소시키고 히알루론산의 합성을 촉진한다는 것을 발견했다. 베르베린은 주름 형성과 관련된 MMP-2 및 MMP-9의 protein 단계 및 mRNA 발현을 감소시키고 TIMP-1 및 TIMP-2의 발현을 증가시켰다. 프로 pro-inflammatory cytokine 으로 알려진 $TNF-{\alpha}$에 대한 베르베린의 억제효능을 실험한 결과, 농도의존적으로 $TNF-{\alpha}$ 유전자를 억제하는 효과를 나타내었다. 이러한 결과를 통해 베르베린은 활성산소종의 억제와 TIMP-1과 -2의 발현 조절에 의한 MMP-2,-9의 감소로 항산화 및 항주름에 효과가 있는 화장품 소재로서의 가치를 가진다고 사료된다.