• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5403-5408
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• 2013

Objective: CXCL12 exerts a wide variety of chemotactic effects on cells. Evidence indicates that CXCL12, in conjunction with its receptor, CXCR4, promotes invasion and metastasis of tumor cells. Our objective was to explore whether the CXCL12-CXCR4 biological axis might influence biological behavior of pancreatic cancer cells. Methods: Miapaca-2 human pancreatic cancer cells were cultured under three different conditions: normal medium (control), medium + recombinant CXCL12 (CXCL12 group), or medium + CXCR4-inhibitor AMD3100 (AMD3100 group). RT-PCR was applied to detect mRNA expression levels of CXCL12, CXCR4, matrix metalloproteinase 2 (MMP-2), MMP-9, and human urokinase plasminogen activator (uPA). Additionally, cell proliferation and invasion were performed using CCK-8 colorimetry and transwell invasion assays, respectively. Results: CXCL12 was not expressed in Miapaca-2 cells, but CXCR4 was detected, indicating that these cells are capable of receiving signals from CXCL12. Expression of extracellular matrix-degrading enzymes MMP-2, MMP-9, and uPA was upregulated in cells exposed to exogenous CXCL12 (P<0.05). Additionally, both proliferation and invasion of pancreatic cancer cells were enhanced in the presence of exogenous CXCL12, but AMD3100 intervention effectively inhibited these processes (P<0.05). Conclusions: The CXCL12-CXCR4 biological axis plays an important role in promoting proliferation and invasion of pancreatic cancer cells.

• Tuberculosis and Respiratory Diseases
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• v.64 no.1
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• pp.22-27
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• 2008

Background: Staphylococcus aureus frequently colonizes and infects hospitalized patients. Respiratory infections with Staphylococcus aureus are common in patients with compromised airway defenses. However the mechanisms of S. aureus invasion from colonization to the epithelium are unclear. Cell invasion by S. aureus would require destruction of the extracellular matrix, which is believed to be the result of increased matrix metalloproteinases (MMP) activity. Methods: In this study, respiratory epithelial cells were infected with S. aureus. After removing the extracellular bacteria by washing, the internalized bacteria in the cells were assessed by counting the colonized forming units (CFUs). The cell adhesion proteins, dysadherin and E-cadherin, were evaluated by Western blotting. The MMPs in the bacterial invasion were evaluated by pretreating the cells with GM6001, a MMP inhibitor. Results: The internalization of S. aureus was found to be both time and dose dependent, and the increase in MMP 2 and 9 activity was also dependent on the incubation time and the initial amount of bacterial inoculation. The invasion of S. aureus was attenuated by GM6001 after 12 hours incubation with a multiply of infection (MOI)=50. The expression of dysadherin, a membrane protein, was increased in a time and dose dependent manner, while the expression of E-cadherin was decreased. Conclusion: MMPs may mediate the invasion of S. aureus into epithelial cells.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.12-25
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• 2003

L-camitine ($\beta$ -hydroxy-${\gamma}$ -trimethyl-ammoniumbutyric acid) is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-camitine, we investigated the effects of in vitro MMP inhibition and activity and expression of UVA-induced MMP 1 in human skin fibroblasts. Fluorometric assays of the proteolytic activities of MMP-l were performed using fluorescent collagen substrates. ELISA (enzyme linked immuno sorbent assay), gelatin-substrate zymography, and RT-PCR ELISA techniques were used for the effects of L-camitine on MMP expression and activity, MMP mRNA expression in UVA irradiated fibroblast. L-camitine inhibited the activities of MMP-l in a dose-dependent manner and the $IC_{50}$/ values calculated from semi-log plots were 2.45mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP expression was reduced 40% by treated with L-carnitine, and MMP-l mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibition the MMP activity, regulation of MMP expression in protein and mRNA level. All these results suggest that L-carnitine may be useful as new anti-aging cofactor for protection against UVA induced MMP expression and activity.

• Ocean and Polar Research
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• v.41 no.2
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• pp.79-88
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• 2019

In this study, the inhibitory effect of Atriplex gmelinii C. A. Mey. against the activity of MMP-2 and MMP-9 secreted from phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 cells was evaluated by gelatin zymography and enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase-chain reaction (RT-PCR), and Western blot assay. Specimens of the halophyte A. gmelinii were extracted twice for 24 hr with methylene chloride ($CH_2Cl_2$), and then twice with methanol (MeOH), in turn. Each extract significantly inhibited the enzymatic activities in gelatin zymography and MMP ELISA kit, and expression of MMP-2 and 9 in mRNA and protein levels. Two crude extracts were combined and then the combined crude extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water ($H_2O$) fractions, according to solvent polarity. Among solvent-partitioned fractions, the 85% aq.MeOH fraction showed the strongest inhibitory effect against MMP-2 and -9 in gelatin zymography and MMP ELISA kit. In RT-PCR, all solvent-partitioned fractions significantly suppressed mRNA expression of MMP-2 and -9. On the other hand, in Western blot assay, all solvent-partitioned fractions except $H_2O$ significantly reduced expression levels of protein. HT 1080 cell migration was most significantly inhibited by the n-BuOH fraction followed by the 85% aq.MeOH and $H_2O$ fractions. These results suggest that A. gmelinii could be used as a potential source to inhibit tumor cell metastasis.

• Journal of dental hygiene science
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• v.14 no.4
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• pp.525-536
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• 2014

Although tobacco use has been known as one of the biggest risk factors on periodontal health, little is known about the effect of smoking cessation on it. The aim of this study was to investigate the change of concentration of matrix metalloproteinase (MMP)-8, MMP-9 and interleukin (IL)-$1{\beta}$ in gingival crevicular fluid (GCF) of 11 quit-smokers for 1 year after smoking cessation. Eleven male subjects to maintain quit-smoking for 1 year participated the oral examination, GCF and saliva collection without periodontal treatments at baseline, after 2 weeks, 2 months, 4 months, 6 months and 1 year. To confirm quit-smoking, nicotine and cotinine concentrations in saliva were measured by high performance liquid chromatography. MMP-8, MMP-9 and IL-$1{\beta}$ concentrations in GCF of upper anterior teeth area were measured by enzyme-linked immunosorbent assay. Change of MMP-8 in GCF during smoking cessation showed fluctuation with decrease (5 subjects) or increase (2 subjects) or maintenance tendency (4 subjects). Changes of MMP-9 were decrease (6 subjects), or increase (2 subjects), or maintenance (3 subjects). Change of IL-$1{\beta}$ also showed fluctuation with decrease (5 subjects) or increase (3 subjects) or maintenance tendency (3 subjects). The subjects with increase tendency had the relatively smaller amount concentration of MMP-8 and MMP-9 at the baseline. It was unclear smoking cessation without periodontal treatment could affect MMP-8, MMP-9, and IL-$1{\beta}$ in GCF. Fluctuation of periodontal biomarkers during smoking cessation might result from feedback interaction between environmental factors and periodontal cells.

• Journal of Life Science
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• v.29 no.3
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• pp.287-294
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• 2019

Calystegia soldanella is distributed in coastal sand dunes and has high environmental adaptability; it is also known to be effective for anti-oxidant, anti-pyretic, anti-septic, and diuretic action. This study investigated the effect of crude extracts and organic solvent fractions of C. soldanella on MMP-2 and MMP-9 expression, MMP activity, and cell mobility in phorbol-12-myristate-13-acetate (PMA)-induced fibrosarcoma HT-1080 cells. C. soldanella was twice extracted, once with methylene chloride (MC) and once with methanol (MeOH). After the MC and MeOH extracts were combined, their suppressive effects on MMP-2 and MMP-9 expression, MMP enzymatic activity, and gene and protein expression were measured by gelatin zymography, enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and western blot method. Cell mobility for the HT-1080 cells was observed by wound healing assay. The combined crude extracts showed a significant suppressive effects on MMP-2 and MMP-9 expression. To explore active inhibitory elements, the combined extracts were fractionated according to polarity into with n-hexane, 85% aqueous methanol, n-butanol, and water. Across these four solvent fractions, MMP-2 and MMP-9 activity and cell mobility in the HT-1080 cells were all strongly inhibited by the n-hexane fraction. These results suggest that C. soldanella extract and organic solvent fractions could be used as potent MMP inhibitors for effective anti-cancer treatments to suppress cancer invasion and metastasis.

• 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.

• Journal of dental hygiene science
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• v.12 no.1
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• pp.71-77
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• 2012

Sex Hormones exert significant influence in body physiology throughout life. Some reports suggested increased sex hormone levels correlate with an increased prevalence of gingivitis. The objectives of this pilot study are to(1) address the link between menstrual cycle and MMP-9, MMP-8, IL-$1{\beta}$ of gingival crevicular fluid during 4 weeks and (2) discuss the major biomarker of periodontal status. 7 female and 3 male volunteer who didn't have smoking habit, medical and dental disease after informed consent, were seen twice a week for 4 weeks. GCF samples were collected and GCF levels of MMP-9, MMP-8, IL-$1{\beta}$ were measured by enzyme-linked immunosorbent assay. The GCF levels of MMP-9, MMP-8, IL-$1{\beta}$ had fluctuation according to menstrual cycle, but the changes of those were not significant by Friedman matched samples test. There is no difference between female and male subjects by Mann-Whitney U test. The correlation of MMP-8, MMP-9 and IL-$1{\beta}$ showed strong by Spearman correlation(0.644-0.945). This study confirms menstrual cycle doesn't influence the periodontium of healthy female subjects.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.101-109
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• 2014

Matrix Metalloproteinases (MMP)-2 and -9 are participated in embryo development, implantation, remodeling of epithelial cell and ovulation. The objective of this study is to evaluate an impact of MMP2 and MMP9 on embryonic developmental competence as well as gene expression profiles of in vitro-produced bovine embryos. After in vitro fertilization, embryos of all groups were transferred into IVC-2 medium treated with MMP2 and MMP9 to check the optimum concentration on the basis of embryo development competence and cell numbers. The optimum concentrations for MMP2 and 9 were 1,200 ng/ml and 300 ng/ml. The blastocyst development competence was not different among 1,200 ng/ml of MMP2 vs. 300 ng/ml of MMP9 vs. combined MMP2 + 9 vs. control groups ($41.46{\pm}10.66$ vs. $37.73{\pm}8.92$ vs. $45.11{\pm}11.41%$ vs. $41.59{\pm}11.88$, respectively). Furthermore, the developmental competences to hatching and hatched blastocysts were not also different among the same groups ($79.84{\pm}12.63$ vs. $83.3{\pm}17.46$ vs. $78.55{\pm}14.48%$ vs. $72.02{\pm}14.09$). In addition, total cell number was significantly (p<0.05) greater in blastocyst treated with MMP9 300 ng/ml among all treatment groups. On the other hand, there was no significant difference of ICM vs. TE ratio in all groups. The expression of five out of six genes (i.e., MMP2, MMP9, IFNt, SSLP1 and HNRNPA2B1) was different among the groups. The expression of IFNt and HNRNPA2B1 genes was significantly greater in MMP9 (p<0.05), but there was no difference of MMP9 expression between MMP2 and MMP9 group (p>0.05). The normalized expression of MMP2 and SSLP1 was greater in MMP2 than other groups (p<0.05). In conclusion, MMPs treatment during IVC-2 medium was remarkably effected on blastocyst developmental competence and gene expression profiles that are related to embryo quality and implantation.

• Applied Biological Chemistry
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• v.47 no.2
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• pp.222-227
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• 2004

Effects of the extracts from bran part of the pigmented rices on inflammation was evaluated by determining their inhibitory action on the production of nitric oxides, histamines and matrix metalloproteinase(MMP) from inflammatory leukocytes. Effects on the production of nitric oxides in a macrophage cell line, RAW264.7 cells, were determined, demonstrating that any significant difference was not detected between the normal rice and the pigmented rice extracts. Inhibitory effects on the histamine-release from a basophilic cell line, RBL-2H3, were examined, showing 3.6 to 5.4-fold increase in the inhibitory activity compared to that of the normal rices. Among the pigmented rice cultivars tested, especially, inhibitory activity of LK1-3-6-12-1-1 was the greatest. Using RAW264.7 cells, we examined the effect of the pigmented rice extracts on the MMP activity. The results showed that the enzyme activity increased with the increasing concentration of the normal rice extract. However, the pigmented rice extracts, except LK1A-2-12-1-l, acted to decrease the MMP activity with their increasing concentrations. The results described above showed the superiority of the pigmented rice extracts in inhibition on release of histamine and MMP, pivotal factors for causing inflammatory responses, from the leukocytes.