• Environmental Mutagens and Carcinogens
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• v.18 no.2
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• pp.98-103
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• 1998

Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

• Imaging Science in Dentistry
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• v.39 no.3
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• pp.121-132
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• 2009

Purpose : To investigate the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of bone sialoprotein (BSP) and osteocalcin (OC) during the differentiation in irradiated MC3T3-E1 osteoblastic cells. Materials and Methods : When MC3T3-E1 osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or $10{\mu}M$ QCT, and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the synthesis of type I collagen, and expression of BSP and OC. Results : The synthesis of type I collagen in cells exposed to 2 Gy of radiation in the presence of 2-DG or QCT showed no significant difference compared with the control group within 15 days post-irradiation. When the cells were irradiated with 8 Gy, 2-DG facilitated the irradiation mediated decrease of type I collagen synthesis, whereas such decrease was inhibited by treating with QCT. During MC3T3-E1 osteoblastic cell differentiation, the mRNA expression of BSP and OC showed the peak value at 14 days and 21 days, respectively. 2-DG or QCT treatment alone decreased the level of BSP mRNA, but increased the OC mRNA level only at early time of differentiation (day 7). In the cells irradiated with 2, 4, 8 Gy, the mRNA expression of BSP and OC decreased at 7 days after the irradiation. The cells were treated with various dose of radiation in the presence of 2-DG or QCT, the mRNA level of both BSP and OC increased although this increase was observed at low dose of radiation (2 Gy) and at the early stage of differentiation. However, when the cells were exposed to 4, 6, or 8 Gy, the increase of BSP and OC mRNAs was detected only in cells co-incubated with QCT. Conclusion : This study demonstrates that 2-DG and QCT affect differently the expression of bone formation related factors, type I collagen, BSP, and OC in the irradiated MC3T3-E1 osteoblasic cells, according to the dose of radiation and the times of differentiation. Overall, the present findings suggest that 2-DG and QCT could have the regulatory roles as radiation-sensitizer and -protector, respectively.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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• pp.69-81
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• 2008

Objective : The purpose of this study was find out the anti-allergic effects of SESHINGO on the allergic rhinitis. Methods : Cell viability was evaluated by the MTT assay. Cells were treated with the indicated concentration of SSG. $TNF-\alpha$ concentration was measured from cell supernatants using ELISA method. IL-4 concentration was measured from cell supernatants using ELISA method.$IFN-\gamma$ concentration was measured from cell supernatants using ELISA method. Total RNA was isolated, $TNF-\alpha$ and IL-4 mRNA expression was detected by RT-PCR analysis Total RNA was isolated, COX-1, COX-2 mRNA was analyzed by RT-PCR analysis. Results : 1. $\beta$-hexosaminidase release in RBL-2H3 cells were decresed by SESHINGO attentively. 2. Secreting ration of $TNF-\alpha$was restrained in RBL-2H3 cells by SESHINGO attentively. 3. Secreting ration of IL-4 was restrained in RBL-2H3 cells by SESHINGO attentively 4. Revelation of $TNF-\alpha$ mRNA was decresed in RBL-2H3 cells as concetration. 5. Revelation of.IL-4 mRNA was decresed in RBL-2H3 cells as concetration. 6. Revelation of COX-2 mRNA was decresed in RBL-2H3 cells. Conclusion : According to above results, to evaluate the effect of SSG on the anti-allergic effects.

• Toxicological Research
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• v.16 no.2
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• pp.95-100
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• 2000

In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.169-173
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• 1995

Border cells are differentiated cells which originate from meristematic cells in The root cap. Experimentally border cells can be released from the root cap by a physical treatment, for example dipping the root tip in the waters After 20-25 hours of release, the new border cell layer forms in the root cap. During the border cell differentiation, new gene expressions were observed in the root cap of pea which was determined by mRNA differential display These new gene expressions may be involved in the border cell differentiation Border cells had unique gene expressions which were determined by mRNA differential display, This suggests that border cells are differentiated cells which are different from the other tissues (ie., leaves, stems, roots or root caps).

• The Korean Journal of Malacology
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• v.16 no.1_2
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• pp.1-9
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• 2000

The visceral ganglion and the right parietal ganglion of the African giant snail, Achatina fulica, consists of two hemispheres, each in left and right side, respectively, like a butterfly. The surface of cortex and medulla in the two ganglions are crowded with nerve cells, but nerve fibers form a network at the middle portion. The nerve cells in the cortex and medulla of the visceral ganglion and the right parietal ganglion are classified into the following four classes according to their sizes: giant (above 200 ${\mu}{\textrm}{m}$, in diameter), large (60-70 ${\mu}{\textrm}{m}$, in diameter), middle (30-40 ${\mu}{\textrm}{m}$, in diameter) and small (10-15 ${\mu}{\textrm}{m}$, in diameter) nerve cells, respectively. The giant and large nerve cells are rarely found(20-22 eas. in total) while the middle and small nerve cells are found in large quantities (middle: 400-500 eas., small: 700-800 eas.). In the AB/AY double staining, the giant nerve cell is identified as light yellow cells (LYC), while large and middle none cells as dark green cells (DGC) or yellow green cells (YGC), and small nerve cells as yellow cells (YC) or blue cells (BC), The DGC, which reacts positively to somatostatin immunostain reaction, inhibits the secretion of the growth control hormone. The giant and large nerve cells are identified to do the functions of phagocytosis as well as neurosecretion.

• Environmental Mutagens and Carcinogens
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• v.24 no.3
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• pp.128-136
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• 2004

CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important. First, we investigated the effect of on CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. We found that B(k)F significantly up-regulates the level of CYP1A1 prompter activity, EROD and CYP1A1 mRNA. When cells were treated with genistein, it was not changed that EROD and CYP1A1 mRNA, compared to that of control. However, genistein inhibited the B(k)F-induced CYP1A1 promoter activity and mRNA level at high concentration. Furthermore, in this study, effects of HDAC(histone deacetvlase) inhibitors on human prostate cancer cells proliferation were examined. HC-toxin, SAHA and TSA inhibited cell proliferation in PC3 cells. A novel HDAC inhibitor, IN2001 also suppressed the growth of PC3 cells. And IN2001 and SAHA increased S phase and G2/M phase at 12 hrs treatment but cells were arrested G0/G1 phase at 45 hrs treatment. The HC-toxin treatment for 24 hrs and 48 hrs increased G0/G1 at low concentration ($0.1\mu\textrm{m}$) but increased G2/M at more than concentration of $1\mu\textrm{m}$. TSA increased G2/M phase. These findings height the possbility of developing HDAC inhibitors as potential anticancer therapeutic agents for the treatment of prostate cancer.

• Journal of Yeungnam Medical Science
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• v.20 no.2
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• pp.129-141
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• 2003

Background: Leptin is a 16-KDa non-glycosylated peptide hormone synthesized almost exclusively by adipocytes. The well-known function of leptin is regulation of food intake and energy expenditure. Leptin also plays a regulatory role in immune and inflammatory process including cytokine production. The purpose of this study was to investigate the effect of leptin on the expression of several chemokine genes(RANTES, IL-8, MCP-1, IP-10, Mig, MIP-$1{\alpha}$, MIP-$1{\beta}$, and GRO-${\alpha}$) in THP-1 cells. Materials and Methods: Total RNA of THP-1 cells were prepared by Trizol method, and then stimulated with the leptin(250 ng/$m{\ell}$) or LPS(100 ng/$m{\ell}$). We examined the expression patterns of various chemokine mRNAs in THP-1 cell lines by RT-PCR and Northern blot. Results: Leptin did not induce the expression of chemokine mRNAs in THP-1 cells. The expression patterns of RANTES, IL-8, MCP-1, IP-10, and Mig mRNAs in THP-1 cells stimulated with leptin and LPS simultaneously was almost same to the patterns of LPS alone-induced chemokine mRNAs. RANTES mRNA expression was independent on the concentrations of leptin. Although leptin did not have strong effect on the expression of RANTES, IL-8, MCP-1, IP-10, Mig, MIP-$1{\alpha}$, MIP-$1{\beta}$, and GRO-${\alpha}$ mRNAs in THP-1 cells, leptin could induce the expression of long isoform of leptin receptor(OB-RL) mRNA, and its expression was elevated in simultaneous stimulation of leptin and LPS. Conclusion: These data suggest that leptin is able to induce OB-RL in THP-1 cells, however, leptin has little effect on the expression of pro-inflammatory chemokine genes.

• The korean journal of orthodontics
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• v.23 no.3 s.42
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• pp.415-425
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• 1993

Fluoride is one of the most potent stimulators of bone formation in vivo. But its direct effects on osteoblast is not yet clear This study was to investigate the effects of Sodium fluoride on alkaline phosphatase(ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in Murin osteoblast-like (MC3T3-E1) cells. The cells were cultured in $\alpha-Minimal$ essential medium $(\alpha-MEM)$ supplemente with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concentration of Sodium fluoride. The ALP activity was assayed by the method of Lowry with disodium phenyl phosphated as substrate. cAMP formation was measured by Radioimmuno Assay(RIA). Type I $\alpha$ 2 collagen ribonucleic acid(mRNA) expression was studied by Nothern blot analysis. The results were as follows: 1. cAMP level was increased by PTH in MC3T3-E1 cells. 2. Sodium fluoride showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-E1 cells. 3. Sodium fluoride increased ALP activity at cocentration of $2{\mu}M,\;4{\mu}M,\;and\;10{\mu}M$ significantly different from control at the 0.001 level. ALP activity revealed maximum value at $10{\mu}M$ in this study. 4. Nothern blot analysis of Sodium fluoride treated cells, using Type I $\alpha$ 2 collagen prove, revealed significant increase at $10{\mu}M$ in MC3T3-E1 cells.

• Environmental Mutagens and Carcinogens
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• v.24 no.3
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• pp.113-120
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• 2004

Chromium compounds are known human and animal carcinogens. In this study, the effects of sodium chromate on apoptosis and cell cycle were investigated in order to unveil the elements of early cellular responses to the metal. Using Chinese hamster ovary cells(CHO-K1-BH4), we found taht chromium (VI) treatment induced apoptosis in these cells, as signified by nuclear fragmentation, DNA laddering on agarose gel electrophoresis, and an increased proportionof cells with hypodiploid DNA. Preceding these changes, chromium (VI) treatment increased caspase 3 pritease activity and also increased expression of p53 protein, while the level of bcl2 protein was not changed. Coincubation with caspase inhibitor, Z-DEVD-FMK, inhibited chromium-induced apoptosis. In the flow cytometric analysis using propidium iodide fluorescence, an increase of cell population in G2/M phase was shown in cells exposed to at least 160 $\mu\textrm{m}$ of sodium chromate for 72h, form 9.8% for 0$\mu\textrm{m}$ chromium (VI) to 26.4% for 320$\mu\textrm{m}$ chromium(VI). Taken together, these findings suggest that chromium(VI)-induced apoptosis is accompanied by G2/M cell cycle arrest, and that p53-mediated pathway may be involved in positive regulation of G2/M arrest and a concurred apoptosis in CHO cells.