• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6245-6248
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• 2013

Aims: To study effects of down-regulation of pituitary tumor-transforming gene (PTTG) on proliferation and metastasis ability of the SCL-1 cutaneous squamous cell carcinoma (CSCC) cell line and explore related mechanisms. Methods: SCL-1 cells were divided into 3 groups (untreated, siRNA control and PTTG siRNA). Cell proliferation assays were performed using a CCK-8 kit and proliferation and metastasis ability were analyzed using Boyden chambers. In addition, expression of MMP-2 and MMP-9 was detected by r-time qPCR and Western blotting. Results: Down-regulation of PTTG could markedly inhibit cell proliferation in SCL-1 cells, compared to untreated and control siRNA groups (P < 0.05). Real-time qPCR demonstrated that expression levels of PTTG, MMP-2 and MMP-9 in the PTTG siRNA group were 0.8%, 23.2% and 21.3% of untreated levels. Western blotting revealed that expression of PTTG, MMP-2 and MMP-9 proteins in the PTTG siRNA group was obviously down-regulated. The numbers of migrating cells ($51.38{\pm}4.71$) in the PTTG siRNA group was obviously lower than that in untreated group ($131.33{\pm}6.12$) and the control siRNA group ($127.72{\pm}5.20$) (P < 0.05), suggesting that decrease of proliferation and metastasis ability mediated by PTTG knock-down may be closely correlated with down-regulation of MMP-2 and MMP-9 expression. Conclusion: Inhibition of PTTG expression may be a new target for therapy of CSCC.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3507-3511
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• 2012

Objective: The aim of this study was to investigate possible mechanisms of LOX gene effects on invasion and metastasis of breast cancer cells by RNA interference. Methods: LOX-RNAi-LV was designed, synthesized, and then transfected into a breast cancer cell line (MDA-MB-231). Expression of LOX, MMP-2 and MMP-9 was determined by real-time PCR, and protein expression of LOX by Western blotting. Cell migration and invasiveness were assessed with Transwell chambers. A total of 111 cases of breast cancer tissues, cancer-adjacent normal breast tissues, and 20 cases of benign lesion tissues were assessed by immunohistochemistry. Results: Expression of LOX mRNA and protein was suppressed, and the expression of MMP-2 and MMP-9 was significantly lower in the RNAi group than the control group (P<0.05), after LOX-RNAi-LV was transfection into MDA-MB-231 cells. Migration and invasion abilities were obviously inhibited. The expression of LOX protein in breast cancer, cancer-adjacent normal breast tissues and benign breast tumor were 48.6% (54/111), 26.1% (29/111), 20.0% (4/20), respectively, associations being noted with clinical stage, lymph node metastasis, tumor size and ER, PR, HER2, but not age. LOX protein was positively correlated with MMP-2 and MMP-9. Conclusion: LOX displayed an important role in invasion and metastasis of breast cancer by regulating MMP-2 and MMP-9 expression which probably exerted synergistic effects on the extracellular matrix (ECM).

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.231-235
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• 2005

Matrix metalloproteinases (MMPs) are a family of structurally and functionally related zinc-dependent enzymes responsible for proteolytic degradation of extracellular matrix components such as base membrane or interstitial stroma. MMPs play an important role in a variety of physiological and pathological tissue remodeling processes, including wound healing, embryo implantation, tumor invasion and metastasis. Since MMP-9 (gelatinase B) has unique ability to cleave type IV collagen, gene expression of MMP-9 has been focused on as a pharmacological target. Flavonoids are a class of compounds that are widely spread in plants. In the coures of screening for the suppressors of MMP-9 gene expression from natural products, Metasequoia glyptostroboides was selected. Six flavonoids, sciadopitysin, isoginkgetin, bilobetin, 2,3-dihydrohinokiflavone, luteolin and apigenin were purified as suppressors of MMP-9 gene expression from M. glyptostroboides. The suppressing activity of the isolated flavinoids on the MMP-9 gene expression was measured by gelatin zymography and Nothern blot analysis.

• Toxicological Research
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• v.14 no.4
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• pp.569-575
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• 1998

One of the most frequent dejects in human cancer is the uncontrolled activation of the ms-signaling pathways. Significant evidence has accumulated to directly implicate members of the matrix metalloproteinases (MMPs) in tumor invasion and metastasis formation. We have previously shown that MMP-9 expression was significantly enhanced in the ras-tranfected HT1080 human fibrosarcoma cells at the mRNA level. In the present study, we investigated the roles of MMP-2 and -9 on the H-ras-induced invasive phenotypes of MCF 10A human breast epithelial cells and HT 1080 human fibrosarcoma cells. We show that H-ras is able to induce or enhance a signaling pathway leading to the enhancement of an invasive phenotype in both MCF10A and HT1080 cells as determined by matrigel invasion assay. We then examined the effect of H-ras activation on the expression of MMP-2 and -9 by measuring enzymatic activities and mRNA levels. Our data clearly demonstrated that H-ras prominently induces expression of MMP-2 in MCF10A cells, while it efficiently up regulates MMP-9 in HT1080 cells. Taken together, these findings suggest that the correlation between ras-mediated invasiveness and enhanced expression of MMPs may be cell type-specific: MMP-9 is closely associated with the invasive phenotype induced by ras activation in fibrosarcoma cells, whereas MMP-2 is more likely associated with it in epithelial cells.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.297-310
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• 2007

치조골 흡수는 파골세포와 matrix metalloproteinases (MMPs)에 의한 골의 무기질과 유기질의 파괴로 일어나는 과정이다. 세균성 산물, 주로 내독소는 치은조직 내에서 염증세포의 유주, 사이토카인 생산, 조직파괴 효소 분비 및 파골세포 활성 등의 국소 면역반응을 유도한다. A. actinomycetemcomitans는 급진성 치주염의 원인 균주중 하나로 그 내독소는 치조골의 흡수와 관련된다. MMP-13은 세균성 산물이나 염증성 사이토카인의 자극에 의해 분비되며, 최근의 연구 결과들은 MMP-13이 치주질환의 진행과 골 흡수 과정에서 일정한 역할을 담당하는 것으로 보고하고 있으나, A. actinomycetemcomitans 내독소와 MMP-13과의 관련성에 대한 연구는 미미하다. 이에 이번 연구에서는 A. actinomycetemcomitans 내독소에 의한 MMP-13의 발현과 파골세포 형성을 세포배양을 통하여 관찰하였고, 백서 구개부 치은에 A. actinomycetemcomitans 내독소를 주입하여 흡수가 진행되고 있는 치조골에서 파골세포의 분화와 MMP-13의 발현을 TRAP 염색, 면역조직화학적 방법 등을 통해 관찰하여 다음과 같은 결과를 얻었다. MMP-13 mPNA의 발현은 A. actinomycetemcomitans 내독소 (1ug/ml)로 24시간 자극한 마우스 치주인대 섬유모세포에서 생리식염수로 자극한 세포에 비하여 약 2.6배 증가하였으며 마우스 대식세포에서는 TRAP 양성 세포가 대조군보다 더 많이 나타났다. A. actinomycetemcomitans 내독소를 주입한 백서 치주조직에서는 대조군보다 더 심한 골소실을 보였다. TRAP-양성 다핵 파골세포 유사세포는 치주염군과 대조군 모두 치조골에서 관찰되었다. TRAP-양성 다핵 파골세포 유사세포는 치주염군에서 대조군보다 유의하게 많은 숫자가 관찰되었으며, 치주염군에서 대조군보다 유의하게 많은 숫자가 관찰되었다. MMP-13 면역양성 반응은 치주염군에서 거친 골연을 갖는 치조골상에 배열된 조골세포와 그 인접한 치주인대에서 관찰되었으며 대조군에서는 MMP-13 면역 양성 반응이 치조골 표면에서만 일부 관찰되었다. 이상의 결과는 A. actinomycetemcomitans 내독속가 MMP-13의 발현을 증가시키며 파골세포의 활성을 통하여 치조골의 흡수를 유도함을 시사한다. 또한 A. actinomycetemcomitans 내독소 투여에 의한 실험적 모델은 백서에서 중등도의 골 소실을 동반한 만성 치주염 모델로 향후 치주질환 치료제의 효과를 평가하는데 유용하게 사용될 수 있으리라 기대된다.

• Biomedical Science Letters
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• v.14 no.3
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• pp.179-186
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• 2008

Matrix metalloproteinases (MMPs), also designated matrixins, hydrolyze components of the extracellular matrix. These proteinases playa central role in many biological processes, such as embryogenesis, normal tissue remodeling, wound healing, and angiogenesis, and in diseases such as atheroma, arthritis, cancer, and tissue ulceration. In previous data, disruption of the actin cytoskeleton by cytochalasin D (CD) inhibited NO-induced apoptosis, dedifferentiation, cyclooxygenase (COX)-2 expression, and prostaglandin $E_2$ production in chondrocytes cultured on plastic or during cartilage explants culture. In this study, we investigated the effects of the actin cytoskeleton architecture on MMP-2 expression and dedifferentiation by CD in rabbit articular chondrocytes. Rabbit articular chondrocytes were prepared from cartilage slices of 2-weeks-old New Zealand white rabbits by enzymatic digestion. CD was used as a disruptor of actin cytoskeleton. In this experiments measuring CD dose response, primary chondrocytes were treated with various concentrations of CD for 24h. The actin disruption was determined by immunostaining. MMP-2 expression levels were determined by immunoblot analysis and Reverse transcriptase-Polymerase chain reaction (RT-PCR) and MMP-2 activity was determined by gelatin zymography. We found that cell morphological change and up-regulation of MMP-2 expression by CD as determined via immunostaining, gelatin zymography and immunoblotting. Moreover, CD induced MMP-2 transcription was detected by RT-PCR. Also, CD-induced type II collagen expression was inhibited by MMP-2 inhibitor I treatment. Our results indicate that CD up-regulated MMP-2 activation causes dedifferentiation of articular chondrocyte.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.335-347
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• 2000

치주질환의 진행에 따른 치주조직파괴에 있어 치주조직내 다양한 세포외기질성분을 분해하는 matrix metalloproteinase-3(MMP-3)는 염증반응에 관여하는 세포들로부터 분비된 interleukin-$1{\beta}$(IL-$1{\beta}$)에 의해 유도될 수 있다. 이전 연구에서 치주인대세포에서의 MMP-3 생성이 tetracycline 및 tetracycline 유도체에 의하여 억제될 수 있음이 보고되었다. 이 연구의 목적은 metronidazole 및 doxycycline-HCl을 적용한 후 치은섬유아세포에 IL $1{\beta}$를 적용하여 MMP-3의 생성을 유도한 후 이들 약물들이 치은섬유아세포의 MMP-3 생성에 미치는 영향을 조사하기 위한 것이다. 건강한 성인으로부터 치주질환이 이환되지않은 상악 제2대구치 후방의 건강한 치은결합 조직을 절취하여 치은섬유아세포를 배양한 후 다양한 농도의 metronidazole (10-$200{\mu}g/m{\ell}$) 및 doxycyline-HCl(10-$200{\mu}g/m{\ell}$)을 각각 적용하여 1시간 배양하고 proMMP-3의 활성화를 유도하기 위하여 25ng/ml의 IL-$1{\beta}$을 투여한 후 24시간 배양하여 배양된 세포의 상층 배양액을 추출하고 proMMP-3 ELISA kit를 이용하여 비색정량하였다. 비색정량을 통하여 얻어진 자료들은 독립 t-test와 일원분산분석(ANOVA) 및 사후검정으로 Duncan test를 시행하여 다음과 같은 결과를 얻었다. 1. Metronidazole 경우 10-$200{\mu}g/ml$의 모든 농도군에서 proMMP-3의 활성도가 억제되었다(p<0.05). 2. Doxycycline-HCl의 경우 $100{\mu}g/ml$ 이하의 농도군에서는 proMMP-3의 활성도가 억제되었으나(p<0.05), $200{\mu}g/ml$ 농도에서는 proMMP-3의 활성도가 상승되었다(p<0.05). 3. Metronidazole과 doxycycline-HCl의 대조군에 대한 각 실험농도군의 proMMP-3 생성의 감소비율 비교시 모든 농도군에서 metronidazole이 doxycycline-HCl보다 더 높은 감소율을 보였다. 이상과 같은 결과는 metronidazole(10-$200{\mu}g/ml$)이 doxycycline-HCl($100{\mu}g/ml$ 이하) 보다 더 광범위한 혈중농도에서 IL-$1{\beta}$의한 인체치은섬유아세포내 MMP-3의 활성도를 효과적으로 억제할 수 있음을 시사하였다.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.85-96
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• 2006

Matrix metalloproteinases (MMPs)는 치주조직내에 존재하는 세포외기질의 유지와 분해에 중요한 역할을 담당하고 있으며 이중 MMP-13은 치주질환의 진행과 깊은 관계가 있다고 알려져 있다. 이번 연구는 치주질환의 진행에 있어서 MMP-13의 활성에 대한 mitogen activated protein(MAP) Kinase의 역할을 구명하기 위해 시행되었다. 백서 치주인대세포에서의 MMP-13 mRNA의 발현은 RT-PCR에 의하여, 그리고 MAP Kinase의 발현은 Western blot에 의하여 측정하였다. $Interleukin-1{\beta}$(IL $-1{\beta}$), Tumor necrosis $factora(TNF-{\alpha})$와 parathyroid hormon(PTH)는 MMP- 13 mRNA 발현을 각각 320%, 180%, 380% 증가시켰으나 bone morphogenetic protein-7(BMP-7)은 MMP-13 mRNA의 발현을 증가시키지 않았다. p38 MAP Kinase 억제제인 SB203580은 IL $-1{\beta}$ 유도 MMP-13의 발현을 약 40% 정도 억제시켰으나, PTH-유도 MMP-13 mRNA의 발현은 억제하지 못했다. IL $-1{\beta}$는 MMP- 13 mRNA의 반감기를 약 2시간 정도로 증가시켰으나, p38 MAP Kinase 억제제로 전처치한 경우에는 반감기가 60분으로 줄어들었다. $IL-1{\beta}$는 p38 MAP kinase와 JNK의 인산화 활성을 증가시켰으나 PTH, $TNF-{\alpha}$와 BMP-7은 p38, JNK, ERK의 활성을 증가시키지 못했다. 이상의 연구결과는 p38 MAP Kinase가 백서 치주인대세포에서의 MMP-13 mRNA 발현을 조절하는데 중요한 역할을 담당함을 시사하였다.

• Genomics & Informatics
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• v.21 no.1
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• pp.9.1-9.13
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• 2023

Matrix metalloproteinase-9 (MMP-9) is a zinc and calcium-dependent proteolytic enzyme involved in extracellular matrix degradation. Overexpression of MMP-9 has been confirmed in several disorders, including cancers, Alzheimer's disease, autoimmune diseases, cardiovascular diseases, and dental caries. Therefore, MMP-9 inhibition is recommended as a therapeutic strategy for combating various diseases. Cinnamic acid derivatives have shown therapeutic effects in different cancers, Alzheimer's disease, cardiovascular diseases, and dental caries. A computational drug discovery approach was performed to evaluate the binding affinity of selected cinnamic acid derivatives to the MMP-9 active site. The stability of docked poses for top-ranked compounds was also examined. Twelve herbal cinnamic acid derivatives were tested for possible MMP-9 inhibition using the AutoDock 4.0 tool. The stability of the docked poses for the most potent MMP-9 inhibitors was assessed by molecular dynamics (MD) in 10 nanosecond simulations. Interactions between the best MMP-9 inhibitors in this study and residues incorporated in the MMP-9 active site were studied before and after MD simulations. Cynarin, chlorogenic acid, and rosmarinic acid revealed a considerable binding affinity to the MMP-9 catalytic domain (ΔG_binding < -10 kcal/ mol). The inhibition constant value for cynarin and chlorogenic acid were calculated at the picomolar scale and assigned as the most potent MMP-9 inhibitor from the cinnamic acid derivatives. The root-mean-square deviations for cynarin and chlorogenic acid were below 2 Å in the 10 ns simulation. Cynarin, chlorogenic acid, and rosmarinic acid might be considered drug candidates for MMP-9 inhibition.

• Korean Journal of Pharmacognosy
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• v.46 no.2
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• pp.154-159
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• 2015

This study was to evaluate the effect of isopropyl alcohol fraction of ultrasonication processed ginseng flower buds(GFB-IF) on the collagen synthesis activity and inhibition of MMP-1 suppression in UV-irradiated human dermal fibroblasts. The higher contents of ginsenoside Rg2(8.234%), Rh1(5.749%), F4(3.881%) in isopropyl alcohol fraction of ginseng flower buds obtained by ultrasonication process at 600W(100℃) for 16 hours. GFB-IF had collagen synthesis effect. GFB-IF induced a significant dose-dependent decrease in the expression for MMP-1 protein. These results suggest that GFB-IF is a potential candidate for the prevention and treatment of wrinkle improving.