• Natural Product Sciences
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• v.2 no.2
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• pp.130-136
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• 1996

Evaluation of plant extracts that might inhibit hepatitis B virus (HBV) replication was performed to find potent anti-HBV agents. Eighty-five species of plants from forty-three families were tested for their anti-HBV activities using HBV-producing HepG2-derived 2.2.15 cells. The anti-HBV activity of plant extracts was measured by slot blot hybridization technique and cytotoxicity was determined by crystal violet staining procedure. All plants were extracted with methanol and the extracts were partitioned into n-hexane, ethyl acetate and aqueous layer. The ethyl acetate fractions of Rhus verniciflua $(stem:\;EC_{50},\;8.2{\mu}g/ml;\;CC_{50},\;9.4{\mu}g/ml)$, Gastrodia elata $(root:\;EC_{50},\;17.7{\mu}g/ml;\;CC_{50},\;>20{\mu}g/ml)$, Raphanus sativus $(seeds:\;EC_{50},\;17.3{\mu}g/ml;\;CC_{50},\;>20{\mu}g/ml)$, and Angelica gigas $(root:\;EC_{50},\;8.3{\mu}g/ml;\;CC_{50},\;15.6{\mu}g/ml)$ revealed the anti-HBV activity in 2.2.15 cell culture system and these fractions are under the process of further sequential fractionation by column chromatography to find the active principles against HBV.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.157-165
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• 1997

포유류 배반포배의 epiblast는 내세포괴에 포함되어 있으며, 이 epiblast cells이 배 및 태아의 생식세포와 일반 체세포로 분화된다 (Beddington, 1986; Lawson 등, 1991). 그런데 조기에 발달된 부화배반포기 배가 지연발생된 부화배반포기 배보다도 많은 epiblast cells을 가지고 있다고 한다(Talbot 등, 1995). 그래서 본 연구에서는 체외수정 유래의 배반포배의 발육속도에 따른 내세포괴/영양배엽세포의 비율 및 배아간세포 확립 효율을 비교하여 발달일령 간에 차이가 있는지를 규명하고자 하였다. 공시한 소의 난포란을 TCM-199에 0.5$\mu\textrm{g}$/ml FSH, 5$\mu\textrm{g}$/ml LH, 10% FBS, 100 units/ml penicilin, 및 100$\mu\textrm{g}$/ml streptomycin을 첨가하여 39$^{\circ}C$, 5% CO2 조건하에서 24시간 동안 체외성숙한 후, 5$\mu\textrm{g}$/ml heparin으로 수정능이 획득된 1$\times$106 sperm/ml의 정자로 체외수정을 유도하였다. 체외수정 후 18~20시간에 과립막세포를 vortexing으로 제거하여 얻은 모든 체외수정란을 3mg/ml BSA, 20${\mu}\ell$/ml NEM amino acids, 40${\mu}\ell$/ml BME amino acids, 10mM glycine 및 1mM alanine이 함유된 CR1aa 배양액에서 BRL 단층세포와 공배양을 실시하였다. 수정후 7, 8 및 9일재 (체외수정일 : 0일)에 확장배반포기까지 발달한 수정란을 이중염색 및 배아간세포의 확립 실험에 공시하였다. 체외수정후 24시간에 분할된 총 1,145개의 수정란이 7, 8 및 9일째에 후기 배반포기까지 각각 222(15.6%), 103(7.2%) 및 52(3.6%)로 발달하여 총 377개 (26.4%)가 발달하였다. 내세포괴/영양배엽세포의 비율은 7일 및 8일째 배반포배에서 각각 47.2$\pm$11.9/95.1$\pm$24.4개 (33/67%) 및 40.3$\pm$12.4/83.3$\pm$26.9개 (33/67%)로서 9일째 배반포배의 19.3$\pm$8.1/62.3$\pm$23.1개 (24/76%) 보다 유의적(P<0.05)으로 높았다. ES-like cells을 확립하기 위하여 후기배반포기 배를 mouse embryonic fibroblast 단층 공배양기에 옮긴 후 5일에 내세포괴의 부착 여부를 판정하고, 10일에 배아간세포의 확립 여부를 판정하였다. 그 결과 7, 8 및 9일째의 배반포기배의 각각 47.7% (82/172), 30.9%(22/71) 및 15.6%(5/32)에서 배아간세포가 확립되었다. 이상의 결과에서 배반포기까지의 발육이 빠른 수정란에서 영양배엽에 대한 내세포괴세포의 비율이 높았고 배아간세포의 확립율도 높다는 사실을 입증할 수 있었으며, 이와 같은 결과에서 체외수정란 유래 배반포배의 질을 결정할 수 있을 것으로 생각된다.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.3
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• pp.189-196
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• 2007

In order to investigate the potential of very low molecular weight hyaluronic acid(oligo HA) as a cosmetic ingredient, we first measured its cytotoxicity in fibroblast, keratinocyte, and SIRC cell lines. For efficacy test, its moisturizing effect and penetration rate were evaluated in an artificial skin system and Caco-2 cells. Oligo HA did not show any cytotoxicity at a concentration of 300 ${\mu}g/mL$ in fibroblasts and 1,000 ${\mu}g/mL$ in keratinocytes but it showed weak proliferation. In vitro ocular test, oligo HA showed negligible cytotoxicity at the maximum concentrations used(2,000 ${\mu}g/mL$) in SIRC cells. In the test of the single and repeated cutaneous applications, oligo HA under occlusive patch did not provoke any cumulative irritation and sensitization. Oligo HA at a concentration of 0.01 % exhibited a more potent moisturizing effect than hyaluronic acid at a concentration of 0.01 %. In the permeability test using artificial skin and Caco-2 cell lines, hyaluronic acid(M.W. $1.1{\times}10^6$) was hardly observed in the down medium of the inserts. On the other hand, oligo HA(M.W. 5,000) was detected in the down medium up to 16.0 % at 6 h in Caco-2 cell culture and up to 90 % at 6 h in an artificial skin system. These results suggest that oligo HA could be useful as an active ingredient for cosmetics.

• Journal of the Korea Convergence Society
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• v.9 no.10
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• pp.93-101
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• 2018

This study was conducted to investigate the anticancer and antitumor effects of quercetin, which is a natural flavonoid substance in human oral cancer cell line HSC-2 cells. The results of this study showed that quercetin reduses the cell viability and the cell proliferation rate, and it led to the evidences of cell death through apoptosis pathway. Also, lower concetration quercetin over $100{\mu}M$ were inhibited the cell migration and invasion. In the present study, we conclude that quercetin treatment of more than $200{\mu}M$ induces apoptosis by activating programed cell death and quercetin treatment of $100{\mu}M$ or more inhibits the cell migration and invasion rate in oral cancer cells. Therefore, this study suggests that quercetin is of sufficient value as an anticancer drug to inhibit metastasis and to treat cancer.

• KSBB Journal
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• v.17 no.2
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• pp.162-168
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• 2002

Human ferritin H- and L-chain genes(hfH and hfL) were cloned into the yeast shuttle vector YEp352 with various promoters, and the vectors constructed were used to transform Saccharomyces cerevisiae 2805. Three different promoters fused to hfH and hfL were used: galactokinase 1 (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase(GPD) promoter and alcohol dehydrogenase 1(ADH1 ) promoter. SDS-polyacrylamide gel electrophoresis and Western blotting analyses displayed expression of the introduced hfH and hfL. In the production of both ferritin H and L subunits GAL1 promoter was more effective than GPD promoter or ADH1 promoter. Ferritin H and L subunits produced in S. cerevisiae were spontaneously assembled into its holoproteins as proven on native polyacrylamide gels. Both recombinant H and L-chain ferritins were catalytically active in forming iron core. When the cells were cultured in the medium containing 10 mM ferric citrate, the cell-associated concentration of iron was 174.9 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L and 148.8 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L but was 49.4 $\mu\textrm{g}$ Per gram(dry cell weight) in the wild type, indicating that the iron contents of yeast is improved by heterologous expression of human ferritin H-chain or L-chain genes.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.239-250
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• 1996

The objective of the present experiments were to determine whether micromanipulative and electro-stimulation conditions for blastomere survival overlapped those for oocyte activation in porcine. Eggs selected for in vitro development potential of blastomeres isolated from 4-cell embryos and oocyte activation by electrostimulation were equilibrated for 5~10 min, in 0.3M sucrose solution containing 7.5$\mu\textrm{g}$/ml cytochalasin B, and then electrostimulated for 30$\mu$sec using one pulse of 100, 120, 150 or 180 volts DC with electrodes 0.2mm apart. Single blastomeres were inserted into empty zona pellucida prior to electrostimulaticn. Then they were cultured in 20${mu}ell$ drops of fresh BECM to observe their developmental ability in vitro in a humidified incubat or at 38.5$^{\circ}C$. The results obtained from these experiments are as follows : 1. When one pulse of 100, 120, 150 or 180 volts DC for 30$\mu$sec were applied to porcine oocytes having the slit formed on zona pellucida for activation, activation rates were 65.1, 66.7, 70.7 and 91.7%, respectively. Higher activation rate was observed in 180V. 2. Infact oocytes incubated for 30 min, in 0.3M sucrose solution after electrostimulation were significantally different from control group with increasing of voltages(p<0.05). When voltages used for electrostimulation were increased, activation rates of oocytes were improved in all treatment groups. 3. When zona punctured-oocytes were only electrostimulated, or incubated in 0.3M sucrose solution for 30 min. after electrostimulation at 180 volt DC, activation rates were 90.5 and 95.5%, respectively. And activation rates of zona punctured-oocytes were significantly different from the groups for which zona pellucida was not punctured(P<0.05). 4. When single blastomeres form 4-cell transferred into empty zona pellucida were incubated for 0, 15 and 30 min. in 0.3M sucrose solution after electrostimulation using one pulse of 180 volt DC for 30 $\mu$sec, developmental rates of electrostimulated-single blastomeres to blastocyst were 72.5, 59.0 and 51.2%, respectively, and the ratio of control group developed to blastocyst were 80.0%. 5. The average cell number in electrostimulated-blastomeres developed to blastocyst were 7.9~10.8, and reduced than the cell number in diploid control ; Also cell number decreased with increasing of voltages. The results of these experiments indicate that the optimal condition for achieving in vitro developmental ability of single 4-cell blastomeres and oocyte activatin is 1 pulse, duration 30 $\mu$sec. in 180 volt, and incubation of blastomeres and oocytes in 0.3M sucrose solution after electrostimulation was not significantally different from another treatment groups. The results also show that this condition is suitable for nuclear transplantation using porcine eggs.

• Journal of the Korean Society of Tobacco Science
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• v.25 no.2
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• pp.128-136
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• 2003

Although tobacco smoke has been known to have genotoxicity as well as cytotoxicity, the sensitivity of the cell lines used against cigarette smoke is poorly understood. The objective of this study was to evaluate and compare the genotoxicity of several cell lines, which are routinely used in the in vitro assays, with cigarette smoke condensate(CSC) of Kentucky Reference Cigarette 1R4F. In the micronucleus(MN) induction assays, murine(CHO-K1, V79, BALB/c 3T3) cell lines and human(MCF-7, A549) ones were used. As a result, the CSC exhibited cytotoxicity with a concentration-dependent response in all cell lines. EC$_{50}$ of CSC in CHO-K1, V79, BALB/c 3T3, MCF-7 and A549 were 140, 125, 100, 116 and 109 $\mu\textrm{g}$/mL, respectively. On the other hand, the spontaneous micronucleated cell(MNC) frequency was stable and reproducible in every cell lines tested in this study. The dose-response of various cell lines to the induction of MN by CSC was estimated using linear regression analysis. CSC(0～100 $\mu\textrm{g}$/mL) caused a dose-dependent MN induction in CHO-K1, V79, BALB/c 3T3 and MCF-7 cell lines. Putting together all the data obtained and linear regression analysis of the data, we concluded that V79 cells are more susceptible to the accurate assessment of CSC-induced MN than the others.s.

• YAKHAK HOEJI
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• v.58 no.1
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• pp.33-39
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• 2014

The objective of this study is to evaluate the anticancer effects of the isoflavone extract from Chungkukjang in human breast cancer, MDA-MB-453 cells. For this study, MDA-MB-453 cells were treated with 12.5, 25, and $50{\mu}g$ isoflavone extract for 24, 48, and 72 hr. Cell proliferations were decreased in a time- and dose-dependent manner. Reduced cell proliferation was suspected by apoptosis or cell cycle arrest. Therefore, after treatment of $50{\mu}g$ isoflavone extract, apoptotic cells were investigated by annexin V staining. The results indicated that isoflavone extract increased the number of early apoptotic cells compared with control. Cleaved PARP was also increased. Next, we investigated the cell cycle and related proteins. The isoflavone extract leads to cell cycle arrest at the G2/M phase. Moreover isoflavone extract had influenced cell cycle relate proteins such as cyclin B1, cyclin A, and p21. These results suggest that isoflavone extract from Chungkukjang induce apoptosis and cell cycle arrest at G2/M phase via regulation of cell cycle-related proteins in MDA-MB-453 cells.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4239-4243
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• 2014

Background: This study investigated the influence of curcumin on HOX transcript antisense RNA (HOTAIR)-mediated migration of cultured renal cell carcinoma (RCC) cells. Materials and Methods: Five RCC cell lines (769-P, 769-P-vector, 769-P-HOTAIR, 786-0, and Kert-3 ) were maintained in vitro. The expression of HOTAIR mRNA was determined by quantitative real-time PCR and cell migration was measured by transwell migration assay. The effects of different concentrations of curcumin (0 to $80{\mu}mol/L$) on cell proliferation was determined by the CCK-8 assay and influence of non-toxic levels (0 to $10{\mu}M$) on the migration of RCC cells was also determined. Results: Comparison of the 5 cell lines indicated a correlation between HOTAIR mRNA expression and cell migration. In particular, the migration of 769-P-HOTAIR cells was significantly higher than that of 769-P-vector cells. Curcumin at $2.5-10{\mu}M$ had no evident toxicity against RCC cells, but inhibited cell migration in a concentration-dependent manner. Conclusions: HOTAIR expression is correlated with the migration of RCC cells, and HOTAIR may be involved in the curcumin-induced inhibition of RCC metastasis.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.15
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• pp.6513-6519
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• 2015

Background: Recent attention on chemotherapeutic intervention against cancer has been focused on discovering and developing phytochemicals as anticancer agents with improved efficacy, low drug resistance and toxicity, low cost and limited adverse side effects. In this study, we investigated the effects of Curcuma C20-dialdehyde on growth, apoptosis and cell cycle arrest in colon and cervical cancer cell lines. Materials and Methods: Antiproliferative, apoptosis induction, and cell cycle arrest activities of Curcuma C20-dialdehyde were determined by WST cell proliferation assay, flow cytometric Alexa fluor 488-annexin V/propidium iodide (PI) staining and PI staining, respectively. Results: Curcuma C20 dialdehyde suppressed the proliferation of HCT116, HT29 and HeLa cells, with IC50 values of $65.4{\pm}1.74{\mu}g/ml$, $58.4{\pm}5.20{\mu}g/ml$ and $72.0{\pm}0.03{\mu}g/ml$, respectively, with 72 h exposure. Flow cytometric analysis revealed that percentages of early apoptotic cells increased in a dose-dependent manner upon exposure to Curcuma C20-dialdehyde. Furthermore, exposure to lower concentrations of this compound significantly induced cell cycle arrest at G1 phase for both HCT116 and HT29 cells, while higher concentrations increased sub-G1 populations. However, the concentrations used in this study could not induce cell cycle arrest but rather induced apoptotic cell death in HeLa cells. Conclusions: Our findings suggest that the phytochemical Curcuma C20-dialdehyde may be a potential antineoplastic agent for colon and cervical cancer chemotherapy and/or chemoprevention. Further studies are needed to characterize the drug target or mode of action of the Curcuma C20-dialdehyde as an anticancer agent.