• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.3757-3761
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• 2012

Objective: Crocin has been proposed as a promising candidate for cancer chemoprevention. The purpose of this investigation was to investigate the chemopreventive action and the possible mechanisms of crocin against human colon cancer cells in vitro. Methods: Cell proliferation was examined using MTT assay and the cell cycle distribution fractions were analyzed using fow cytometric analysis after propidium iodide staining. Apoptosis was detected using theTUNEL Apoptosis Detection Kit with laser scanning confocal microscope. DNA damage was assessed using the alkaline single-cell gel electrophoresis assay, while expression levels of p53, cdk2, cyclinA and P21 were examined by Western blot analysis. Results: Treatment of SW480 cells with crocetin (0.2, 0.4, 0.8 mmol/L) for 48 h signifcantly inhibited their proliferation in a concentration-dependent manner. Crocetin (0.8 mmol/L) signifcantly induced cell cycle arrest through p53-independent mechanisms accompanied by P21 induction. Crocetin (0.8 mmol/L) caused cytotoxicity in the SW480 cells by enhancing apoptosis and decreasing DNA repair capacity in a time-dependent manner. Conclusions: This report provides evidence that crocetin is a potential anticancer agent, which may be used as a chemotherapeutic drug.

• The Journal of Internal Korean Medicine
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• v.21 no.1
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• pp.65-73
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• 2000

Objectives : The effects of cotreatment of adriamycin and ethanol extract of herb (Palgin-tang hab Hwajuck-hwan a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origin cell lines, Chang. Methods : Chang(ATCC) liver cells were cultured in RPMI-1640(Gibco SRL Co, Gaithersburg, MD) badge including 10% fetal bovine serum. Chang liver cells were treated with various concentrations(from 10 to $0.16{\mu}l$) of adriamycin and herb extract(from 500 to $31.25{\mu}l$) After 48h later, the cells were tested for viability by Crystal violet staining assay. Adriamycin and Herb extract induced ladder pattern of DNA fragmentation in Chang cells. Genomic DNA was isolated and separated on 1.5% agarose gels. The DNA was stained with ethidium bromide and visualized under UV light. Results : The death of Chang cells was synergistically induced by the cotreatment of adriamycin and ethanol extract of herb. In addition, the cotreatment-induced cell death of Chang cells was mediated by apoptotic death signal processes. The phosphotransferase activity of JNK1 remained in a basal level in Chang cells which was treated individually with the adriamycin and ethanol extract of herb. However, it was markedly increased in Chang cells which was cotreated with adriamycin and ethanol extract of herb. In addition, the expression of Fas and FasL was markedly induced by the cotreatment of adriamycin and herb extract. For a while, the expression of Sax was a eminently increased by the ethanol extract of herb. However, Scl2 expression was not affected by the individual or cotreatment of adriamycin and herb extract. Conclusions : our results suggest that the cotreatment of adriamycin aM ethanol extract of herb induces synergistic apoptotis of human liver origin Chang cells via the upregulation of JNK, Fas, FasL and Bax.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.9
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• pp.1143-1150
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• 2017

The aim of this study was to determine the chemical and intracellular antioxidant activities of ${\beta}$-carotene and lycopene and to compare their quantitative structure-activity relationship (QSAR). In our previous study, the second ionization energy of lycopene was higher than that of ${\beta}$-carotene, as calculated by QSAR. Chemical antioxidant activities of ${\beta}$-carotene, lycopene, and Trolox were examined by measuring ferric reducing antioxidant power (FRAP) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Intracellular antioxidant activities were evaluated by intracellular reactive oxygen species (ROS) and DNA fragmentation. The FRAP of lycopene was higher than that of ${\beta}$-carotene (P<0.05), and the two carotenoids had similar antioxidant activities in DPPH radical scavenging activity assay. Trolox had the greatest chemical antioxidant activities (P<0.05). When RAW264.7 cells were treated with lipopolysaccharide (LPS) (100 ng/mL) for 20 h, intracellular ROS and DNA fragmentation significantly increased (P<0.05). RAW 264.7 cells pretreated with ${\beta}$-carotene ($4{\mu}M$) and lycopene ($0.4{\sim}2{\mu}M$) for 4 h formed significantly less intracellular ROS than LPS-treated control cells (P<0.05), whereas cells with Trolox did not reduce production of intracellular ROS. In addition, cells pretreated with $2{\mu}M$ lycopene produced less intracellular ROS than those treated with ${\beta}$-carotene (P<0.05). DNA fragmentation of cells with ${\beta}$-carotene and lycopene was similar to that of LPS-treated control cells as measured by Hoechst staining. The antioxidant ability of lycopene was greater than that of ${\beta}$-carotene in the QSAR, FRAP, and intracellular ROS assays (P<0.05). ${\beta}$-Carotene and lycopene had lower antioxidant activities as measured by FRAP (P<0.05) but higher intracellular protective effects against LPS-induced oxidative stress in comparison with Trolox.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.419-429
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• 2000

Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

• Journal of Life Science
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• v.19 no.6
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• pp.787-792
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• 2009

In this study, protease-producing bacteria were isolated from the marine sedimentary layer in coastal Jeju. We isolated 2 protease producing strains (SK-2 and SK-125) and tested their protesase producing activities. Gram staining and BIOLOG of isolated strains revealed that strains SK-2 and SK-125 belong to Bacillus and Pseudoalteromonas families, respectively. The 16S rDNA nucleotide sequences analyses of the isolated strains showed 99% sequence homology with those of Bacillus sp. and Pseudoalteromonas sp.; therefore, the isolated strains SK-2 and SK-125 were named Bacillus sp. SK-2 and Pseudoalteromonas sp. SK-125, respectively. The optimum conditions for the cell growth of protease activities were obtained when the both isolates were cultured at $30^{\circ}C$, 96 hrs and pH $7{\sim}8$.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.181-186
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• 2007

Inhibitor of DNA binding protein or inhibitor of differentiation(Id) is largely considered as positive and/or negative regulators of proliferation, differentiation, angiogeneisis, and apoptosis. The four Id genes(Id1, Id2, Id3, and Id4) were known in mammals. However, little is known about the expression and function of these genes in reproductive physiology. Among them, this study was conducted to analyze the expression pattern of Id3 mRNA on folliculogenesis in rat ovary. After PMSG administration, the ovaries were obtained at 3, 6, 12, 24, 36, and 48hrs, fixed, dehydrated, and paraffin embedded. For in situ hybridization, anti-sense and sense Id3 cRNA probes were prepared and applied to the ovarian section. The ovarian sections were coated with NTB-2 emulsion. After that, the slides were developed and counterstained with hematoxylin and eosin staining. The hybridization signal was estimated on a scale of 1+ to 4+. In oocyte, the intensity of Id3 mRNA in primordial and primary follicles was scored at ${\geq}2+$, but the intensity was less than 1+ in secondary, dominant, and preovulatory follicles. In granulosa cells, the Id3 mRNA was strongly expressed(3+ or 4+) in dominant and preovulatory follicles. Taken together, Id3 mRNA was expressed specifically at follicle stages and follicular tissue and might be closely related with follicle development.

• The Korean Journal of Food And Nutrition
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• v.28 no.6
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• pp.956-964
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• 2015

Probiotics is known improve the microenvironment of colon; however, the metagenomic DNA study of its lactic acid bacteria in constipation induced by loperamide is not clearly understood. In the present study, we investigated the reduction of the lactic acid bacteria in case of constipation, in normal and loperamide-induced rat. Lactic acid powder (lactic acid bacteria 19) was prepared from Chong Kun Dang Pharmaceutical Corporation. After 2 weeks of oral administration, the group treated with the higher concentration of lactic acid bacteria ($10^9CFU/mL$ per kg of body weight) following loperamide treatment was the most effective in increasing number, weight, and water content of feces. A similar but significant increase was found in the group treated with lower concentration of lactic acid bacteria ($10^7CFU/mL$ per kg of body weight) after loperamide treatment. The concentrations of acetic acid and propionic acid in feces in the loperamide-induced rat with high concentration lactic acid, were significantly higher than that of others. Furthermore, gastrointestinal transit ratio as well as the length and area of intestinal mucosa were significantly increased after treatment with lactic acid bacteria in loperamide-induced rat. Metagenomics DNA analysis indicated that the microorganism homology in cecum was similar between the groups of normal (NOR) and HIG. Our results show that lactic acid bacteria were effective in improving the constipation.

• Polymer(Korea)
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• v.32 no.1
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• pp.26-30
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• 2008

This study was designed to investigate the effect of hybridization of synthetic/natural materials for annulus fibrosus (AF) tissue regeneration in vitro and in vivo. The synthetic/natural hybrid scaffolds were prepared using PLGA (poly (lactic-co-glycolic) acid), SIS (small intestinal submucosa) and DBP (demineralized bone particles). PLGA, PLGA/SIS(20%), PLGA/DBP(20%) and PLGA/SIS (10%)/DBP (10%) scaffold were manufactured by solvent casting/salt leaching method. Compressive strength was measured. Rabbit AF cells were isolated, cultured and seeded into experimental groups. Hydroxyproline production and DNA quantity of AP cells on each scaffold was measured at 2, 4 and 6 weeks after in vitro culture. Cell-scaffold composites were implanted subcutaneously into athymic mice. After 1,4 and 6 weeks postoperatively, specimens were taken and H&E, Safranin-O and type I collagen staining were carried out concerning formation of cartilagenous tissue. In vitro PLGA/SIS scaffold was evaluated for total collagen content (bydroryproline/DNA content) and PLGA scaffold was evaluated for compressive strength.

• Korean Journal of Veterinary Research
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• v.44 no.3
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• pp.379-387
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• 2004

When an organism is exposed to various toxicants chronically, reactive oxygen species(ROS) are accumulated and eventually result in several biological effects from gene expression to cell death. In the present study we investigated the oxidative damage of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin(TCDD) and/or benzo(a)pyrene (B(a)P) in C100 cells. C100 cells treated with TCDD(30 nM) and B(a)P($3{\mu}M$) underwent diverse oxidative stress as determined through thiobarbituric acid-reactive substances(TBARS) formation, DNA fragmentation, DNA single strand break(SSB) assay, immunohistochemical staining of 8-hydroxy-2'-deoxyguanosine(8-OHdG), and mRNA expressions of antioxidant enzymatic genes such as Cu/Zn-SOD gene, GPx(glutathione peroxidase 5) gene, and catalase gene. Lipid peroxidation in C100 cells was determined through measuing the formation of TBARS. For theat, the cells were pretreated with TCDD(30 nM) and/or B(a)P($3{\mu}M$) for 0.5, 1, 2 and 4 days. TBARS formation was increased in TCDD(30 nM) and B(a)P($3{\mu}M$) and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) and positive control treatment groups comparing to the controls. Mixture treatment induced more DNA fragmentation than the single treatment group at day 6. Also, SSB in all treatment groups was clearly observed when compared with the negative control group. As with the expression of antioxidant enzyme, GPx 5mRNA, B(a)P alone and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) treatment were higher comparing to those of the negative control and TCDD treatment groups. Our results suggest that exposure of C100 cells to mixture of TCDD and B(a)P leads to significant oxidative damage comparing to the exposures to the individual chemicals. Mechanisms of action are discussed. Additional studies are needed to elucidate the detailed mechanism of mixture-induced toxicity.

• Journal of Life Science
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• v.24 no.9
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• pp.1012-1018
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• 2014

Sasa quelpaertensis Nakai (Korean name, Jeju-Joritdae) is one of the most abundant plants on Mt. Halla, Jeju Island, and it has long been used in traditional medicines. Recent studies have reported it as possessing various beneficial functions, including anti-inflammatory, anti-diabetic, anti-hypertension, anti-gastritis, anti-oxidant, and anti-cancer effects. However, the molecular mechanisms of its anti-cancer activity have not been clearly elucidated. In this study, we investigated the anti-cancer effects and mechanism of S. quelpaertensis on human colon cancer HT-29 cells. Cell growth inhibition by S. quelpaertensis was determined by MTT assay. Apoptosis was performed by DNA fragmentation, flow cytometry with propidium iodide staining (PI), and reverse transcription-polymerase chain reaction (RT-PCR) to confirm the anti-apoptotic factors, such as inhibitor of apoptosis (IAP) family members. $NO^{\bullet}$ production was determined by Griess assay. S. quelpaertensis treatment resulted in the time- and dose-dependent inhibition of the cell viability of HT-29 cells by inducing apoptosis, as evidenced by the accumulation of the sub-G1 cell population stained by PI, as well as the ladder-like DNA fragmentation in a dose-dependent manner. S. quelpaertensis-inducing apoptosis was accompanied by the induction of S cell cycle arrests, increasing $NO^{\bullet}$ concentrations, and the down-regulation of IAPs, including X-chromosome-linked IAP (XIAP), cellular IAP-1 (cIAP-1), cIAP-2, and survivin. Taken together, these findings have important implications for future clinical developments of S. quelpaertensis in colon cancer treatment.