• Journal of Life Science
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• v.14 no.2
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• pp.263-268
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• 2004

The sprouting of vascular endothelial cells is an initial step in angiogenesis. Matrix metalloproteinases can associate with integrin on the surface of endothelial cells, thereby promoting angiogenesis. The purpose of this study was to test if fibroblast growth factor-2 (FGF-2) can regulate the sprouting in porcine pulmonary artery endothelial cells. FGF-2 induced sprouting and secretion of MMP-2 and plasmin. FGF-2 also induced the expression of integrin Mac-1, which is inhibited IS20I. Addition of BB-94, a 2-antiplasmin and IS20I inhibited FGF-2-induced sprouting activity. Therefore, FGF-2-induced sprouting activity in PPAECs may be accomplished by secretion of proteinases such as MMP-2 and plasmin and integrin expression.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1229-1234
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• 2006

This study investigated whether matrix metalloproteinases (MMPs) can be modified in apoptotic smooth muscle cell (SMC) using the SMC that undergoes apoptotic death by expressing Fas-associated death domain containing protein (FADD) when they are grown without tetracycline in culture medium. In the absence of tetracycline, FADD-SMC lost adherence and showed the fragmentation of the nuclei. In proportion to duration of tetracycline removal, phosphorylated form of p38 MAPK and of ERK increased, whereas phosphorylation of protein kinase B (PKB) was not changed very much in response to tetracycline The levels of cyclin A and cyclin D were also decreased in a time dependent manner. Up-regulation of MMP-9 expression and activity was observed when the SMC were grown without tetracycline. Immunoreactivity of MMP-9 was detected from both attached and floating FADD-SMCs grown without tetracycline. An inhibitor of MAPK kinase, PD098059, and an inhibitor of p38 MAPK, SB203580, inhibited the up-regulation of MMP-9. Treatment of the SMC with a synthetic MMP inhibitor, BB94, attenuated death occurring in the absence of tetracycline. These results indicate that SMC undergoing death is able to up-regulate MMP-9 and that the enzyme can affect cell viability.

• Journal of Life Science
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• v.26 no.10
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• pp.1189-1195
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• 2016

Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) are potent angiogenic factors that have been used clinically to induce angiogenesis. To enable migration and invasion, cells must proliferate and secrete proteinases, which degrade the surrounding extracellular matrix. The goal of this study was to investigate the cell proliferation; matrix metalloproteinase-2 (MMP-2), MMP-9, and plasmin secretion; and migration and invasion of glioma-derived U-373-MG cells induced by VEGF and HGF treatment. An additional goal was to test the hypothesis that elevated secretion of MMP-2, MMP-9, and plasmin contributed directly or indirectly to the proliferation, migration, and invasion of U-373-MG cells. Cell proliferation, migration, and invasion and MMP-2, MMP-9, and plasmin secretion were significantly increased in the VEGF and HGF-treated U-373-MG cells. To elucidate the role of the increased secretion of MMP-2, MMP-9, and plasmin in cell proliferation, migration, and invasion of the U-373-MG cells, they were treated with MMPs inhibitor (BB-94) and plasmin inhibitor (α2AP) prior to VEGF or HGF stimulation. The BB-94 and α2AP treatment resulted in a significant reduction in the cell proliferation, migration, and invasion of the U-373-MG cells as compared with the VEGF- and HGF-treated groups. The results indicate that inhibition of MMPs and plasmin reduce the cell proliferation, migration, and invasion of U-373-MG cells.

• 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.

• Journal of Oral Medicine and Pain
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• v.26 no.2
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• pp.95-105
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• 2001

제 4형 콜라젠을 분해하는 MMP-9은 조직 재생에 중요한 역할을 하기 때문에 주목받아 왔는데, 이전의 문헌들에서 PMA에 의해 이 유전자 발현이 강하게 상승발현된다고 알려져 있다. 비록 MMP-9 발현을 조절하는 PMA의 기전이 잘 밝혀지지는 않았지만, 다른 유전자발현에서의 이 phorbol ester의 효과는 c-raf-1-ERK 신호전달통로의 활성에 관해 연구되어 오고 있다. 하지만 이번 연구에서 저자는 MMP-9 발현에서 PMA의 상승효과에는 대개는 스트레스성 자극과 관련된 JNK1 의존성 신호전달과정이 또한 필요하다는 다른 가능성을 조사하였다. 그 결과 JNK 억제재중 하나인 SP 600125가 UM-SCC-1세포주에서 PMA에 의해 유도된 MMP-9 상승발현을 용량의존적으로 억제하는 것으로 나타났고 72시간동안 전처치를 한 경우에 그의 억제효과가 최대이었다. phorbol ester로 처리한 세포에 GAL4 luciferase reporter와 vector를 주입해서 조사한 결과 PMA가 c-Jun transacting 활성을 상승시켰다. 그뿐 아니라 PMA에 의한 MMP-9 촉진자 활성에는 AP-1 motif가 필요하며 이 motif에 c-Jun이 결합하는 것을 EMSA를 통해 확인하였다. 결론적으로 UM-SCC-1 세포주에서 PMA에 의한 MMP9의 증가된 발현은 이미 밝혀진 ERK 경로뿐 아니라 JNK 경로를 통한 결과임이 밝혀져 이 경로를 차단하는 방법이 또하나의 암치료 방향을 제시해주고 있다.

• KSBB Journal
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• v.24 no.5
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• pp.425-431
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• 2009

Hepatocyte growth factor (HGF) and its receptor play an important role in the formation and progression of glioma. In this study, I investigated the ability of HGF to recover of the PSA siRNA-suppressed cell proliferation, migration and invasion in U-251-MG cells. PSA siRNA-transfected U-251-MG cells showed the reduction of the proliferation, migration and invasion with compared to control. Treatment of HGF on the PSA siRNA-transfected U-251-MG cells recovered the ability of proliferation, migration and invasion. These data suggest that PSA and HGF may use unique and parallel signaling cascade leading to the proliferative, migrative and invasive phenotype of U-251-MG cells. I also showed that PSA cooperated with HGF to a migrative and invasive phenotype via the increased secretion of matrix metalloproteinase-2 (MMP-2) and MMP-9.

• Journal of Life Science
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• v.28 no.3
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• pp.293-299
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• 2018

Viticis fructus (fruits of Vitex rotundifolia) is the dried fruit from Vitex rotundifolia; is a traditional medicine for treating inflammation, migraines, chronic bronchitis, headaches, eye pain, and gastrointestinal infections; and demonstrates various bioactivities, including anti-allergic, anti-cancer, and anti-inflammatory effects, which are partly due to its phenolic compound content. This study examines the inhibitory effects of viticis fructus (fruits of Vitex rotundifolia) on MMP-2 and MMP-9 expression using gelatin zymography and RT-PCR in phorbol-12-myristate-13-acetate (PMA)-induced HT-1080 fibro-sarcoma cells. Fruits of Vitex rotundifolia were extracted twice using dichloromethane ($CH_2Cl_2$) and methanol (MeOH). The combined crude extracts ($CH_2Cl_2$ and MeOH) significantly inhibited MMP-2 and MMP-9 activities in gelatin zymography. The combined extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq. MeOH), n-butanol, and water, successively according to polarity. Among all solvent-partitioned fractions, 85% aq. MeOH fractions showed the strongest inhibition on the activation of MMP-2 and MMP-9 in gelatin zymography. In PMA-stimulated HT-1080 cells, the expression levels of MMP-2 and MMP-9 mRNA were also greatly inhibited by the 85% aq. MeOH fraction. These results suggest that viticis fructus can be used as an excellent source for anti-invasive agents.

• Journal of Life Science
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• v.22 no.9
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• pp.1254-1260
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• 2012

The moringa (Moringa oleifera Lam.) plant is used both as food and an anti-allergic agent. In this study, we investigated skin protection effects of methanol extracts from the root, seed, fruit, and leaves of moringa in HaCaT keratinocyte cells. To investigate the pharmacological potential of various moringa extracts on TNF-${\alpha}$-induced collagen degradation in HaCaT cells, we measured the activity of matrix metallopeptidase-9,2 (MMP-9,2) by zymography analysis. Our results showed that all the moringa extracts inhibit the TNF-${\alpha}$-induced enzyme activity of MMP-9. In particular, moringa root extracts significantly suppressed MMP-9 and MMP-2 in a dose-dependent manner. Next, to investigate the anti-inflammation effect of the moringa extracts, we examined cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and interleukin-6 (IL-6) expression of the extracts. The results showed that both the root extracts and the seed extracts decreased the TNF-${\alpha}$-induced expression of COX-2. In addition, the root and leaf extracts reduced the expression of IL-6. However, none of the moringa extracts affected the expression of iNOS. The results suggest that moringa root extracts down-regulate MMP-9, COX-2, and IL-6 and that the root extracts offer superior skin protection effects compared with other extracts of moringa in HaCaT cells.

• Tuberculosis and Respiratory Diseases
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• v.56 no.3
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• pp.289-296
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• 2004

Background : Mucus hypersecretion in the patients with airway diseases represents poor prognosis as well as discomfort. However, there is no known therapy for its effective control. One important component of mucus is mucin, a glycosylated protein, which endows mucus with viscosity. We studied whether a proteinase has a role in mucin secretion and if so, which. Methods : (1) Inhibition of mucin secretion Group-specific proteinase inhibitors were tested to evaluate whether a proteinase belonging to a group of proteinases plays a role in mucin secretion. Phenylmethylsulfonyl fluoride(PMSF, a serine proteinase inhibitor), E-64(a cysteine proteinase inhibitor), Pepstatin(an aspartic proteinase inhibitor) and 1, 10-Phenanthroline(a metalloproteinase inhibitor) were treated into the Calu-3 cell line for 24 hours. The enzyme linked immunoabsorbant assay(ELISA) for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with a control. (2) Stimulation of mucin secretion Matrix metalloproteinase-9(MMP-9), MMP-12 and TACE(TNF-alpha converting enzyme), which are known to be related with airway diseases, were used to be treated into Calu-3 for 24 hours. ELISA for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with the controls. Results : (1) PMSF($10^{-4}M$), E-64($10^{-4}M$), Pepstatin($10^{-6}M$) and 1, 10-Phenanthroline($10^{-4}M$) reduced the MUC5AC secretion by $1{\pm}4.9%$(mean${\pm}$standard deviation; P=1.0 compared with the control), $-6{\pm}3.9%$(P=0.34), $-13{\pm}9.7%$(P=0.34) and $41{\pm}8.2%$(P=0.03), respectively. (2) The amounts of MUC5AC secretion stimulated by MMP-9(250ng/ml), MMP-12(100ng/ml) and TACE(200ng/ml) were $103{\pm}6%$(P=0.39), $102{\pm}8%$(P=1.0) and $107{\pm}13%$(P=0.39), respectively, compared with the controls. Conclusion : Metalloproteinase(s) is (are) suggested to play a role in mucin secretion. It appears that metalloproteinases, other than MMP-9, MMP-12 or TACE, affect the mucin secretion in this in vitro model.

• KSBB Journal
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• v.18 no.6
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• pp.485-489
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• 2003

Hepatocyte growth factor (HGF) is a mesenchymal-derived cytokine. It exerts a motogenic effect on various target cells, which is displayed either by cell scattering, locomotion, and migration during the wound repair process of cultured cells, or invasiveness through the extracellular matrix, in vitro. Although it is known that HGF influences the motogenic effect of endothelial cells, the precise effects of HGF during migration are still poorly understood. To elucidate the role of HGF in endothelial cell migration, the effect of HGF on endothelial cell migration and MMPs and plasmin production were studied. We found that HGF induces the migration of cultured endothelial cells through increased MMPs and plasmin secretion.