• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3519-3528
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• 2012

Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1998.11a
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• pp.93-96
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• 1998

NAPL이 다핵방향족화합물의 하나인 phenanthrene 의 생분해에 미치는 영향을 알아보았다. Pseudomonas putida CRE7 을 이용한 실험에서 NAPL 의 첨가로 인한 가장 큰 차이는 미생물의 소수성의 변화였다. 소수성이 증대됨으로써 phenanthrene 의 가용성이 증대되었으며, 이로 인해 더 많은 양의 오염물 분해가 이루어졌다. 생물학적 분해의 관찰은 발생되어지는 $^{14}$ $CO_2$의 radioactivity 측정을 통해 이루어졌으며, 미생물의 소수성 측정은 bacterial adhesion to hydrocarbon (BATH) assay 를 이용하였다.

• International Journal of Oral Biology
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• v.30 no.1
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• pp.31-37
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• 2005

Treponema maltophilum, a Group IV oral spirochete, is associated with periodontitis and endodontic infections. In this study we analyzed the functional role of the major surface protein of this organism (MspA) in human gingival fibroblasts (HGFs). The full-length gene encoding MspA was cloned and expressed in Escherichia coli by using the expression vector pQE-30. The recombinant protein (rMspA) was purified by affinity chromatography with nickel-nitrilotriacetic acid agarose and possible contamination of E. coli endotoxin in rMspA was removed by using polymyxin B-agarose. rMspA significantly induced the expression of pro inflammatory cytokines like IL-6 and IL-8 and intercellular adhesion molecule (ICAM)-1 in HGFs, when analyzed by reverse transcription-PCR, flow cytometry, and enzyme-linked immunosorbent assay. Our results indicate that MspA of T. maltophilum may play an important role in amplifying the local immune response by upregulating the expression of proinflammatory cytokines and ICAM-1.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.491-498
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• 2011

During mammalian fertilization, germ cell-specific hyaluronidases, such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5), are important for the dispersal of the cumulus mass. In this study, we demonstrated that bull Hyal5 is a single copy gene on chromosome 4 that is expressed specifically in the testis. In addition, we expressed recombinant bull SPAM1 and Hyal5 in human embryonic kidney 293T cells and showed that these enzymes possessed hyaluronidase activity. We also demonstrated that a polyclonal antibody against bull sperm hyaluronidase inhibits sperm-egg interactions in an in vitro fertilization (IVF) assay. Our results suggested that bull Hyal5 may have a critical role in bull fertilization.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3383-3387
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• 2015

Portulaca oleracea (Family: Portulacaceae), is well known for its anti-inflammatory, antioxidative, anti-bacterial, and anti-tumor activities. However, cytotoxic effects of seed oil of Portulaca oleracea against human liver cancer (HepG2) and human lung cancer (A-549) cell lines have not been studied previously. Therefore, the present study was designed to investigate the cytotoxic effects of Portulaca oleracea seed oil on HepG2 and A-549 cell lines. Both cell lines were exposed to various concentrations of Portulaca oleracea seed oil for 24h. After the exposure, percentage cell viability was studied by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT), neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed a concentration-dependent significant reduction in the percentage cell viability and an alteration in the cellular morphology of HepG2 and A-549 cells. The percentage cell viability was recorded as 73%, 63%, and 54% by MTT assay and 76%, 61%, and 50% by NRU assay at 250, 500, and $1000{\mu}g/ml$, respectively in HepG2 cells. Percentage cell viability was recorded as 82%, 72%, and 64% by MTT assay and 83%, 68%, and 56% by NRU assay at 250, 500, and $1000{\mu}g/ml$, respectively in A-549 cells. The 100 $100{\mu}g/ml$ and lower concentrations were found to be non cytotoxic to A-549 cells, whereas decrease of 14% and 12% were recorded by MTT and NRU assay, respectively in HepG2 cells. Both HepG2 and A-549 cell lines exposed to 250, 500, and $1000{\mu}g/ml$ of Portulaca oleracea seed oil lost their normal morphology, cell adhesion capacity, become rounded, and appeared smaller in size. The data from this study showed that exposure to seed oil of Portulaca oleracea resulted in significant cytotoxicity and inhibition of growth of the human liver cancer (HepG2) and human lung cancer (A-549) cell lines.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.293-301
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• 2004

1. 연구 목적 Fibronectin은 세포외기질의 주요성분인 거대 당단백질로서, 조골세포의 부착과 증식 및 이동능에 중요한 역할을 담당한다고 알려져 있다. 이러한 fibronectin의 조골세포에 대한 영향을 실제 임상에 적용하기 위해서, 전체 fibronectin 단백질을 사용하는 것은 면역학적으로나 경제적으로 많은 단점을 안고 있어서, 유효한 반응단위만을 추출하여 활용하는 것이 바람직한 방법으로 알려져 있다. 이 연구의 목적은 세포부착에 주로 관여하는 fibronectin type III분절 중 10번 도메인이 조골양 세포에 미치는 영향을 전체 fibronectin단백질과 fibronectin type III 7-10 도메인 분절과 비교, 관찰하는 것이다. 2. 연구 방법 사람의 fibronectin을 기초로 한 적절한 primer로서, 유전자 재조합법을 이용하여 fibronectin type III 10 도메인과 fibronectin type III 7-10 도메인 분절을 얻었으며, 전체 fibronectin분자는 상용으로 준비하여 24-well 세포배양 용기에 도포하였다. 배양된 조골양세포(HOS cell)를 $1x10^5$ cells/well의 농도로 각 well에 분주하여 $37^{\circ}C$에서 1시간 배양을 하였다. Cell adhesion assay를 실시하기 위해 10% formaldehyde로 고정시키고 1% Crystal Violet으로 염색하여 광학현미경을 관찰 후 2% SDS를 처리하여 microplate reader기를 이용하여 570nm에서 혼탁정도를 측정하였다. 음성대조군으로는 RPMI 용액을 사용하였다. 동일한 방법을 이용하여 준비된 $35mm^2배양접시에 HOS cell을 $37^{\circ}C$에서 4일간 배양 후, MTS assay를 이용하여 세포 증식도에 미치는 영향을 관찰하였다. 6일째 405nm에서 활성화된 세포에서 분비된 p-nitrophenol을 이용한 alkaline phosphatase activity를 측정하였다. 3. 결과 및 고찰 Fibronectin type III 10 도메인은 HOS cell에 대한 생물학적인 효과면에서, 전체 fibronectin 분자 및 fibronectin type III 7-10 분절과 통계적으로 유사한 세포부착도를 보여주었으며, 세포증식도와 alkaline phosphatse 활성도면에서도 큰 차이가 나타나지 않았다. 이상의 연구결과로 볼 때, fibronectin type III 10 도메인이 조골세포의 증식을 목적으로 사용하는 생체재료의 표면개질 부착물질로 응용할 수 있는 가능성이 있다고 하겠다.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.2
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• pp.115-123
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• 2006

Objective: To investigate new signal transduction cascade through integrin, FAK and ERK in the suppressed cell proliferation by GnRH-I and -II. Method: Human endometrial cancer cells (HEC1A) were cultured under the following condition: DMEM/F12 (10% FBS). GnRH-I and -II were treated time (0, 5, 10, 15, 20, 30 min; 100 nM) and dose (10 nM or 100 nM; 20 min) dependent manner according to experimental purposes. Cell proliferation was measured using [$^3H$] thymidine incorporation assay. Immunoblotting was utilized to detect proteins. Results: GnRH-I and -II inhibited proliferation of HEC1A cells and induced expression of integrin ${\beta}3$. Phosphorylation of FAK and ERK were induced by GnRH-I and -II. Conclusion: GnRH inhibited cell proliferation via the expression of integrin and FAK, ERK phosphorylation.

• The Korea Journal of Herbology
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• v.24 no.3
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• pp.39-50
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• 2009

Objectives : Smilacis glabrae rhizoma (SG) has been traditionally used as a herbal medication of musculoskeletal disorders like arthritis, pain, convulsions, and syphilis in traditional Korean medicine. This study was investigated anti-oxidative and anti-inflammatory effect of fractionated extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (HUVEC). Methods : SG extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of SG onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymethoxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of ICAM-1 and VCAM-1 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially ethyl acetate (EA) extract, significantly inhibited free radical generation, the TNF-$\alpha$-induced intracellular oxidation. Furthermore, the EA extract protected TNF-$\alpha$-induced adhesion to THP-1, expression of adhesion molecules accompanied by an attenuation of IL-6 and IL-8 formation in HUVEC. Conclusions : These results indicate that EA extract of SG have potential as an agent of atherosclerosis and other chronic inflammatory diseases including diabetes, hypertension, and arthritis.

• Food Science of Animal Resources
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• v.35 no.4
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• pp.551-556
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• 2015

The aim of this study was to evaluate the functional properties of lactic acid bacteria from various sources and to identify strains for use as probiotics. Ten Lactobacillus strains were selected and their properties such as bile tolerance, acid resistance, cholesterol assimilation activity, and adherence to HT-29 cells were assessed to determine their potential as probiotics. Lactobacillus sp. JNU 8829, L. casei MB3, L. sakei MA9, L. sakei CH8, and L. acidophilus M23 were found to show full tolerance to the 0.3% bile acid. All strains without L. acidophilus M23 were the most acid-tolerant strains. After incubating the strains at pH 2.5 for 2 h, their viability decreased by 3 Log cells. Some strains survived at pH 2.5 in the presence of pepsin and 0.3% bile acid. Lactobacillus sp. JNU 8829, L. acidophilus KU41, L. acidophilus M23, L. fermentum NS2, L. plantarum M13, and L. plantarum NS3 were found to reduce cholesterol levels by ＞50% in vitro. In the adhesion assay, Lactobacillus sp. JNU 8829, L. casei MB3, L. sakei MA9, and L. sakei CH8 showed higher adhesion activities after 2 h of co-incubation with the intestinal cells. The results of this comprehensive analysis shows that this new probiotic strain named, Lactobacillus sp. JNU 8829 could be a promising candidate for dairy products.

• Korean Journal of Materials Research
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• v.20 no.6
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• pp.312-318
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• 2010

Multilayer Poly methyl methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates were fabricated using electrospinning and in vitro investigations were carried out for pre-clinical biocompatibility studies. The initial cellular cytotoxicity of the methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay using fibroblast-like L-929 cells. Cellular adhesion and differentiation studies were carried out using osteoblast-like MG-63 cells. As simulated body fluid (SBF) contains the same ionic concentration of body fluid and any bioactive material tends to deposit bone-like apatite on the samples surfaces into the SBF, in vitro bioactivity of the multilayer bone plates were investigated using SBF. We also studied the internal organization and tensile strength of the multilayer PMMA/PVA bone plates using micro-computed topography (${\mu}$-CT) and universal testing instrument (UTI, Korea) respectively. The cellular cytotoxicity study with MTT confirmed that the cellular viability was 78 to 90% which indicates good cyto-compatibility. Scanning electron microscopic findings revealed a good attachment and adhesion phenomenon of MG-63 cells onto the surfaces of the samples. Cellular differentiation studies also showed that osteogenic differentiation was switched on in a timely manner and affirmed along with that of the control group. Bone-like apatite formation on the surfaces was confirmed within 14 days of SBF incubation. Initial organizations of the multilayer PMMA/PVA bone plates were characterized as dense and uniform. The tensile strength of the post-pressing electronspun mat was higher than that of the pre-electronspun mat. These results suggest that a multilayer PMMA/PVA bone plate system is biocompatible, bioactive and a very good alternative bone plate system.