• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5391-5396
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• 2013

Aim: To investigate the role of Golgi phosphoprotein 3 (GOLPH3) in tumour growth and metastasis of esophageal squamous cancer. Methods: A lentiviral shRNA-vector was utilized to stably knockdown GOLPH3 in Eca-109 esophageal squamous cancer cells. mRNA transcription and protein expression of GOLPH3 were examined by real-time quantitative PCR and Western blotting, respectively. Cell proliferation activity was assessed by MTT assay and invasion and migration potentials by matrigel invasion and transwell motility assays. Results: Stable knockdown in the GOLPH3 cell line was established. PD-A gene expression was significantly suppressed by lentivirus-mediated RNAi, which resulted in reducing the capacity for cell proliferation, migration, invasion and adhesion in vitro. In vivo, GOLPH3 depletion resulted in inhibition of tumour growth, with stable decrease in the expression of GOLPH3 in tumor xenografts. Conclusions: Our findings suggest that lentivirus mediated silencing of the GOLPH3 gene has a significant anti-tumour effect on esophageal squamous cancer in vitro and in vivo. In addition, the results indicate that GOLPH3 might be an effective molecular target for gene therapy in esophageal squamous cancer.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.525-528
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• 2003

Spatholobus Suberectus dunn. has been applied to blood stasis in oriental medicine. We selected one potent ethyl acetate subfraction-2 from Spatholobus Suberectus Dunn.(SSD) from anti-metastasis screening. It exerted the cytotoxicity against HT1080 and B16BL6 with the IC50 of 60 ug/ml and also significantly inhibited tumor cell induced platelet aggregation (TCIPA). It effectively didn't inhibit cell adhesion of HT1080 to matrigel coated wells, while it inhibited the cell invasion of HT1080 at the doses of 10, 20, 40 μg/ml by Boyden chamber assay. It effectively suppressed lung metastasis by B16BL6 melanoma in C57BL6 mice. These results indicate that the EtOAc subfraction-2 of Spatholobus Suberectus Dunn. can be applied to caner treatment with anti-metastatic activity.

• Journal of Microbiology and Biotechnology
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• v.28 no.8
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• pp.1332-1338
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• 2018

In the course of studies to discover natural products with anti-angiogenic properties, two cyclic octapeptides, octaminomycins A (1) and B (2), were isolated from the cultures of Streptomyces sp. RK85-270. Octaminomycins suppressed the vascular endothelial growth factor (VEGF)-induced proliferation, adhesion, tube formation, migration, and invasion of HUVECs. Anti-angiogenic activity was futher confirmed in vivo by the chicken chorioallantoic membrane assay. We also identified that 1 and 2 inhibited the phosphorylation of VEGF receptor 2, AKT, and ERK1/2 and the expression and activities of MMP-2 and MMP-9. These results suggest that 1 and 2 may serve as potential scaffolds for the development of therapeutic agents to angiogenesis-dependent diseases.

• Journal of Haehwa Medicine
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• v.10 no.2
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• pp.21-29
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• 2002

To evaluate the antitumor activity and antimetastatic effects of Polygoni Orientalis Fructus(POF), studies were done experimentally. The results were obtained as follows : 1. In cytotoxicity against A549, and XF498 cell concentration inhibiting cell growth up to below 30% of control was recognized at $200{\mu}g/m{\ell}$ of POF. Also POF inhibited cell growth up to below 30% of control against SK-OV-3, SK-MEL-2, HCT15 and MCF-7 cell at $100{\mu}g/m{\ell}$. 2. The concentration inhibiting adhesion of A549 and SK-OV-3 to complex extracellular matrix up to below 70% of control was recognized at $100{\mu}g/m{\ell}$ of POF. 3. In pumonary colonization assay, a number of colonies in the lungs were decreased significantly in POF treated group as compared with control group. These results suggested that POF extracts might be usefully applied for prevention and treatment of cancer.

• Nutrition Research and Practice
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• v.17 no.5
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• pp.844-854
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• 2023

BACKGROUND/OBJECTIVES: Fucoidan, a polysaccharide content in brown algae, has been reported to inhibit the growth of cancer cells. The present study aimed to investigate the suppression effects of fucoidan on A549 non-small cell lung cancer cells migration. MATERIALS/METHODS: The anti-migratory activity of fucoidan in A549 cells was examined by wound healing assay and phalloidin-rhodamine staining in response to fucoidan (0-100 ㎍/mL) treatment for 48 h. Western blot analysis was performed to clarify the protein expressions relevant to migratory activity. RESULTS: Fucoidan (25-100 ㎍/mL) significantly suppressed A549 cells migration together with reduced the intensity of phalloidin-rhodamine which detect filopodia and lamellipodia protrusions at 48 h of treatment. The protein expression indicated that fucoidan significantly suppressed the phosphorylation of focal adhesion kinase (FAK), Src, and extracellular signal-related kinase (ERK). In addition, the phosphorylation of p38 in A549 cells was found to be increased. CONCLUSIONS: Our data conclude that fucoidan exhibits anti-migratory activities against lung cancer A549 cells mediated by inhibiting ERK1/2 and FAK-Src pathway.

• Tuberculosis and Respiratory Diseases
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• v.52 no.2
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• pp.145-155
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• 2002

Background: Inflammation, where vascular endothelial cells are activated by cytokines, recruits circulating leukocytes such as neutrophils into the tissues. Mononuclear phagocytes as well as tissue cells activated by these stimuli produce these chemokines. In this study, thr effects of IL-1 and LPS on the expression of CXC chemokines such as GRO-${\alpha}$, IL-8 and ENA-78 in vascular endothelial cells and the neutrophil adhesion effects of ENA-78 and GRO-${\alpha}$ was investigated. Methods: Human umbilical vein endothelial cells were cultured and stimulated with various concentrations of IL-1 and LPS. The concentrations of the GRO-${\alpha}$, IL-8 and ENA-78 secreted were measured using enzymelinked immunosorbent assay. The effects of ENA-78 and GRO-${\alpha}$ on neutrophil adhesion to the endothelial cells were also investigated. Results: The addition of IL-1 and LPS to the vascular endothelial cells induced GRO-${\alpha}$ IL-8 and ENA-78 secretion in a time- and dose-dependent manner. The neutrophil adhesion was also increased by induction of ENA-78 and GRO-${\alpha}$ to the vascular endothelial cells in a dose-dependent manner. Conclusion: CXC chemokines such as GRO-${\alpha}$, IL-8 and ENA-78 secreted by the vascular endothelial cells play an important role in the acute inflammatory responses by stimulating neutrophil adhesion to the vascular endothelial cells, raising the possibility that the CXC chemokines are one of the targets in the clinical application of acute inflammation.

• Biomedical Science Letters
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• v.2 no.1
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• pp.31-40
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• 1996

Escherichia coli is one of the most common etiological agents in urinary tract infection. An important virulence factor is the adhesive capacity of E. coli to uroepithelial cell, mediated by bacterial fimbriae. The Adhesion property has been regarded as an important virulence determinant in urinary tract infections. A total of 60 patients, who were diagnosed microbiologically as urinary tract infections, were examined by immunoblotting and enzyme-linked immunosorbent assay(ELISA). Uropathogenic E. coli with recombinant plasmid were positive for mannose resistant hemagglutination (MRHA). For identification of p-fimbriae subtype in uropathogenic E. coli, In the immunoblot analysis, specific bands in the range of p-fimbriae molecular weight of 17KD-22KD were identified. For the distribution of p-fimbriae subtype in the patient sera, 34/60(56.7%) were positive for $F7_1$, 28/60(46.7%) were positive for $F7_2$, and 30/60(50%) were positive for F13 with immunoblotting method. similar trends were observed in the enzyme-linked immunosorbent assay. Relatively good specificity(92.6%) and sensitivity(90%) were found in the ELISA test system using mixed antigens of purified $F7_1$, $F7_2$, and F13 p-fimbriae, and 60 sera from patients with urinary tract infections. In conclusion The serological tests were convenient method in diagnosis of urinary tract infections. among those ELISA could be recommended in diagnosis of urinary tract infections.

• Tuberculosis and Respiratory Diseases
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• v.48 no.6
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• pp.887-897
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• 2000

Background : Since the exact pathogenesis of sepsis-induced ARDS has not been elucidated, the mechanisms of enhanced neutrophilic respiratory burst were probed in endotoxin primed neutrophils associated with the roles of phospholipase A2(PLA2), platelet activating factor(PAF) and apoptosis. Methods : In isolated fresh human neutrophils, effects of the inhibition of PLA2 and PAF on the apoptosis were examined by the method of Annexin-FITC/dual PIflow cytometry. The roles of PLA2 and PAF on the neutrophilic respiratory burst were also examined by measuring oxidant generation in cytochrome-c reduction assay. Activities of the PLA2 and lysoPAF acetyltransferase (lysoPAF AT) of the neutrophils were determined to understand the effect of endotoxin on these enzymatic activities which may be related to the neutrophilic respiratory burst and apoptosis. In addition, the role roles of PLA2 and PAF in neutrophilic adhesion to bovine endothelial cells were examined in vitro by neutrophil adhesion assay. To investigate the effect of oxidants on pulmonary surfactant, cytochemical ultrastructural microscopy was performed. To inhibit PLA2 and PAF, non-specific PLA2 inhibitor mepacrine (100 nM) and WEB 2086 (100 nM) or ketotifen fumarate (10 ${\mu}g$/ml) were used respectively in all in vitro experimental sets. WEB 2086 is PAF receptor antagonist, and ketotifen fumarate is a lyso PAF AT inhibitor. Results: The mapacrine treatment, provided and the endotoxin (ETX) treatment, resulted in increased apoptosis of neutrophils (p<0.001) while treatments of WEB 2086 and ketotifen did not. The inhibition of PLA2 and PAF decreased (p<0.001) production of oxidants from PMA-stimulated neutrophils. While endotoxin increased the PLA2 activity of neutrophils (p<0.01), mepacrine supressed (p<0.001) the activity, provided after treatment of ETX. The lyso PAF actyltransferase activity (lyso PAF AT) increased (p<0.01) after treatment of ETX. In contrast, mepacrine, WEB 2086 and ketotifen showed a tendency of decreasing the activity after treatment of ETX. The treatment of ETX incresed (p<0.001) neutrophil adhesion to endothelial cells, which was reversed by inhibition of PLA2 and PAF (p<0.01). The binding of oxidants to pu1monary surfactant was identified histologically. Conclusions : The enhanced neutrophilic respiratory burst by ETX plays a pivotal role in the pathogenesis of ARDS in terms of oxidayive oxidative stress. Increased production of oxidants from neutrophils is mediated by the activations of PLA2 and lyso PAF AT.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.3
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• pp.199-218
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• 2005

Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.5
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• pp.581-585
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• 2005

Five lactic acid bacteria (LAB) including 2 Lactobacillus strains isolated from Kimchi were evaluated to determine the binding ability to Caco-2 cells and $AFB_1$. LAB were divided into three different groups ; viable, heat-treated, and acid-treated cells. In the radioactive-labeling assay for bound cell counting, viable Lactobacillus Plantarum KCTC 3099 showed the higher adhesion to Caco-2 cells with the binding capacity of $39.2\%$, which was $149\%$ higher than Lactobacillus rhamnosus GG as a positive control. Leuconostoc mesenteroids KCTC 3100 showed the similar binding ability to L. rhamnosus GG. After 1 hour incubation at $37^{\circ}C$ with $AFB_1$, viable L. Planterum KTCC 3099 removed the toxin by $49.8\%$, which was similar level to L. rhamnosus GG. Both heat- and acid-treated groups showed high binding effect but acid-treated group was more effective for both Caco-2 cell binding and $AFB_1$ removal than the other. These results indicate that components of bacterial cell wall might be involved in tile binding to intestinal cells and toxins.