{pA4-5] [ 10/19/2000 (The) 10:00 ~ 11:00 / {Hall 8) } CARCINOGENESIS AND ITS CHEMOPREVENTION IN EXPERIMENTALLY ~ INDUCED GASTROESOPHAGEAL REFLUX DISEASE. Jeong-Sang Lee!, Tae-Young Oh?, Byoung-Ok Ahn®, Hyun Cho®, Young-Joon Surh! and Ki-Balk Haim? ‘college of Pharmacy. Seoul National University: 2ong~A Pharmaceutical Research Institute: Spepertment of Gastroenterology. Alou University School of Medicine, Korea. Gastroesophageal reflux disease (GERO) is multifactorial in etiology and is characterized by movement of acid and other noxious substances from the stomach into the esophagus. The most severe histologic consequence of chronic gastroesophgeal reflux is Barrel's esophagus, which has been considered as a premalignant condition often leading to the formation of adenocarcinoma of esophagus. Oxidative DNA damage and subsequent mutation may contvibute to oncogenesis related to GERD. In order to catiy the role of oxidative stress in pathogenesis of GERD and Barrett's esophagus, we developed an animal model and Investigated the possible protective effacts of selected antioxidative substances. For this purpose, Sprague-Dawiey rats ‘were subjected to duodenal ligation using a small-lumen sing to provoke GERD. Two days after the operation, the malondialdehyde formation in GERD esophagus was significantly increased with Concomitant reduction in cellular reduced-alutathione levels, which correlated well with histologic severity. Increased levels of indicibe nitric oxide synthase and cyclooxygenase-2 were also observed. Nuclear extracts from the esophagus of GEAD rats exhibited dramatic activation of the ruclear transeription factor, NF-KB, which was essociated with increased k~a degradation and subsequent nuclear translocation of the p65 subunit. Oral administration of the gastrorotective drug ranitidine or of DA-3601 (100 mg per kg) derived from the medicinal plant Artemisia asiatica ‘markedly attenuated not only histologic abnormalities and formation of reactive oxygen species but also activation of NF=«B in GER animals. [pA4-8] [ 10/19/2000 (The) 10:00 - 11:00 / {Hall 8} Genotoxic Study of Kamijadowhan in In Vivo Supravital Staining and tk+/ - Gene Assays Kim JA°, Kwon 0S, Pyu JC Toxicology Laboratory, Korea Institute of Sclence end Technology, Seoul, 136-850 Karijadowhan (KM), one of the oriental medicine, has been reported to have a potent anti~ metastasis effect. With the purpose of developing new anti~metastasis drug through the collaboration with other groups, the evaluation of KMJ toxicity was made Ia our lab. KM toxicity was assessed using the tk"/“forward gene mutation assay in L5178Y mouse lymphoma cells and in vivo supravital cytogenetic assay in mice. In the tk*/~forward gene mutation assay with L5178Y mouse lymphoma cell, KMJ reveaied no significant increase of MF (mutation frequency) that was observed in the presence and absence of S~9 metabolic activation systoms at various ‘concentrations of KMJ (313, 625, 1250, and 2500 s@/). In the supravital staining micronucieus. assay with mouse perioherai reticulocytes. no significant increase of micronucleated reticulocytes (MNRETs) was observed atter a single intraperitoneal administration of KIM (2500, 1250 and 625, 16/'@) to mice. This data indicate that MNRETS in mouse reticulocytes and mutations in the tk*!~ forward gene assay are not induced by KML. [PAd~7] { 10/19/2000 (The) 10:00 ~ 11:00 / {Hall 8} ) 161The Inhibitory Effects of Houttuynia cordata THUNB against Cadmium induced Cytotoxicity (1!) Lee JH1, You 182, Kim JS2, Lee KN3, Chung WY4, Han DS4, Back SHi# 1Dept. Natural Products and aDept. of Industrial Oriental Medicina, Professional Graduate Schoo! of Oriental Medicine. 40ent, of Oral Anatomy, School of Dentistry, Wonkwang University. ‘2Dent. of Industrial Chemistry Iksan College, iksan, Korea, ABSTRACT—This study was conducted to investigate the antitoxic component in aqueous extract ‘of Houttuynie cordata THUNB, The results were as follows: General, detoxication effects by Houttuynia cordate THUNB extract increased in proportion to the extract concentrations in rats. ‘When 40 mg/kg dosage of Houttuynia cordeta THUNB extract was administrated, Houttuynia cordata THUNS extract showed the highest antitoxic effects in metalothionein induction. After the extract treatment, body weights increased in proportion to the extract concentrations. However, after 3 weeks, the body weight decteased insignificantly. From the above results, Houttuynia ‘cordate THUNG extract increased metallothionein concentration and decreased the toxicity of ‘cadmium in rats. In vitro the antitoxic activity of aqueous extract of Houttuynia cordata THUNB on NIH 273 fibroblasts was evaluated by the MTT (3-(4,5-dimethythiazo!~2-yl)~2,5~ diphenyi-2H- tetrazollumbromie) and SRB (sulfocnodamine 8 protein) assays, The light microscopic study was Cattied out to observe morphological changes of the treated cells, These results were obtained as follows: The concentration of 10-2 mg/ml of Houttuynia cordata THUNE extract was shown significant anttoxic activity. The number of NIM 37S fibroblasts were increased and tend to Tegenerate, These results suggest that Houttuynia cordata THUNB extract retains 2 potential antitoric activity. [PA4-8] [ 10/19/2000 (Thr) 10:00 = 11:00 / (Hall 6) ] ‘The Inhibitory Effects of Trichosanthes kirllowil against Cadmium induced cytotoxicity ( Ill) Lee JH, You St, Kim Sk2, Lee KN3, Chung WY4, Han OS4, Back SH Dept. of Natural Products & 3Dept. of Industrial Oriental Medicine, Professional Graduate Schoo! ‘of Griental Mecicine, 4Dept. of Orat Anatomy, Wonkwang Univ. {Oept. of Industrial Chemistry ‘22Dept. of Environmental Horticulture, san College. korea Abseract~ This study was conducted to investigate the antitxic agents in aqueous extrat of ‘Trichosanthes kiilowi. The results were as follows: Generally. dotoxitication effects by ‘Tichosanthes kirlowi extract increased in propartion to the extract concentration in rats. When 40 ma/kg dosage of Trichosanthes Kirlowi extract was administrated, Trichosanthes kirlowil extract Showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body \weights increased in proportion to the extract concentrations. From the above results, ‘Trichosanthes Kirlow! extract increased methallothionein concentration and decreased the toxicity cof cadmium in rats. In vito the antitoxic activity of aqueous extract of Trichosanthes kirlowii on NIH 379 fibroblasts was evaluated by the MTT (3-(4,5-dimethyithiazo!~2-y1)~2.5~diohenyl- 2H- tetrazolium bromide) and SRE (sulforhodamine 8 protoin)assays. The light microscopio study was Carried out to observe morphological changes of the treated cells. These results were obtained as follows: The concentration of 10-2 mg/ml of Trichosantnes kiriowil extract was shown sighiticant antitoxic activity. The number of NIH &T3 fibroblasts were increased and tend to regenerate. These Tesuits suggest thet Trichosanthes Kirlowii extract retains a potential antitoxic activity [PA4-8] [ 10/19/2000 (Thr) 10:00 ~ 11:00 / [Hall 8) } Protective Effects of Butanol Fraction of Carthamus tinctorius L. Semen on 162Endotoxin Induced Thrombosis in Rats ‘Seung Keum-Aan,Baik Sue-Jeong.Juna Joo-Hee,Joo Kyeoung-Mi,Kang Hye-Kyung. Kim Young ‘MinJeong Choon-Sike,Jung KI-Hwa College of Pharmacy. Ouksung Women's University ‘The concems about diseases of a cardiovascular system have increased with the rise of living standard and the trend of advanced age. Among these, the thrombus causes the serious dis ‘ike apoplexia, cerebri and myocardial infarction. Thrombosis is caused by the injury of fendothalium and the alteration in normal blood flow. Endotoxin is the product from gram negative bacteria, and protein-lipo~ polysaccharide is known as a main component of it. its used in experiment for antithrombosis activity screening In our study, we injected endotoxin(4000EU/kg, iv.) in rats at thr after administration of Carthamus tinctorlus L. Semen butanol fraction(50Gmg/kg, 0.0.). To investigate activities of Carthamus tinctorlus L. Semen butanol traction for biood coagulation system, we measured blood cloting time, prothrombin time, florinogen and fibrinogen degradation products in vivo, antiplatelet garegation activity and the stabilizing effect on heat-induced hemolysis in vio. And then we measured superoxide dismutase acitivity, glutathione content, glutathione S-transferase activity and malon dialdenyde content to figure out the mechanism of anticoagulation. [AS a result, Carthamus tinctorius L. Semen butanol traction has antiplatelet aggregation activity /n vitro, delays blood clotting ime and prothrombin time, and decreases fibrinogen and fitxinogen degradation products in vivo. Also, it increases superoxide dismutase activity, glutathione content and glutathione S-transferase activity, and decreases maion dialdehyde content. On the basis of our study, we may propose that a blood coagulation system and cell inury is suppressed by the antithrombosis effect of Carthamus tinctoris L. Seren. EpAs-10) [ 10/19/2000 (The) 10:00 ~ 11:00 / [Hall 8) ] Analysis of Benzophenone and 4-Nitrotoluene in Water, Sediments and Soils by Gas Chromatography/Mass Spectrometry hee HK, Kwon OS, Ayu JC Toxicology Lab.. Korea Institute of Science and Technology. P.O.Box 131, Cheongryang, Seoul 180-650, Korea The benzophenone (8°) and 4-nitrotoluene (4-NT) classified as endocrine disrupting chemicals were determined in water, sediment and soll. The modified SPEED9B method for water samples ‘and ultrasonic extraction of US EPA (method 35508) method for sediment and soll samples were sed for the analysis of 8P and 4-NT. n-Hexane was used for the extraction of BP and 4-NT in the water, sediment and soil samples. 2uf of the concentrated solution (0.3 m of final volume) \was applied to GC/MSD. The method detection limits af BP were 10 ng/tfor water samples, and 0.25 ng/g and 1 ng/g for sediment and solls. For 4=NT, method detection limits were 5 n/t for water samples and 1 ng/o for sediment and soil samples. As a result. BP concentrations were ranged from 24.4 ng/t to §3.6 ng/tat 7 sites of water samples and from 10.3 ng/Z to 13.9 n/t at 2 sites of soll samples. wnich were higher than those of water and soll Danks. 4~NT was not elected In water, sediment and soll samples. {PA4=11] { 10/19/2000 (The) 10:00 ~ 11:00 / (Hall 8] 1 Effect of glycolic acid alone or combination with UVB on skin iritation and inflammation in guinea pigs Hong JT. Park KS®, Kim HJ. Kim EJ, Juno KM, Ahn KS, Lee JK, Man KT. Kim OJ, Kim YK and Lee 163SH National institute of Toxicological Research, Korea Food and Drug Administration ‘The use of alpha-hydroxy acids(AHAs) containing cosmetics has aroused the public interest with their supposed abil to reduce wrinkles, roughness, age spots of skin and other sions of sunburn ‘damages. The excessive and chronic use of AHA containing cosmetics could cause skin iitation, sswolling and sunburn, and may increase photo-toxicily and photo-carcinogenesis. However, exact dose-response relationship, photo-toxic effects and skin toxic mechanisms have not been known, In the present study, dose and time effects of glycolic acid, one of the most commonly used ‘AAs, alone or combination with UVB on skin iritation and inflammatory response were examined Skin ititation by glycolic acid and UVB alone was increased in dose and time-dependent manners. Higher dose of alyoolic acid and UVB (3 J/em2) treatment for 2 weeks caused severe skin Intation. Lower dose of glycolic acid and UVB (0.4 J/cm2) caused slight or mild irritation. However, lower glycolic acid enhanced UVB-induced skin irtation resulting in severe iritation, Histological examination showed that glycolic acid dose dependently reduced integrity of stratum Ccorneun and increased skin thickness, and higher dose of glycolic acid destroyed epidermal layer without inflammatory response. UVB icreased skin thickness, and caused condensed inner stratum comeum and reduced its integrity of outer layer. Glycolic acid enhanced UVB~induced the reduction of stiatum coreum integrity. Completely lost of organization of stratum coreum was, seen in UVB and glycolic acid combination treated skin. Glycolic acid did not change basal or UV-induced PGE2 production and COX-2 protein expression. UVB, whe‘eas.increased PGEZ (60% over contro! by higher dose of UVB) and COX-2 expression(2 and 3 fold). These results show that glycolic acid cause skin iritation in a dose and time dependent manners and enhance UvB- Induced skin iitation, however glycolic acid~induced skin iritation may not be associated with inflammatory response. [A412] [ 10/19/2000 (Thr) 10:00 ~ 11:00 / {Hall 8) } Co-carcinogenic potential of glycolic acid in hairless mouse skin Hong JT®, Ahn KS, Kim EJ, Jung KM, Park KS, Lee JK. YP Yune. YK Kim, KW Ha and Lee SH National Institute of Toxiological Research, KFA and College of Pharmacy Chungbuk National University Alpha-hydroxy acids (AMAs) are organic acids present in naturel sources such as frults, wine and milk. Such sources of AHAs have been used as cosmetic material for several years in the public wit their supposed abiity to reduce wrinkles. roughness, age spots of skin and other signs of sunburn damages, However. It is also true that the excessive and chronic use of AHAS containing ‘cosmetics could cause skin iitation, swelling, sunbum, photo-toxicty, and that increase of pphote-carcinogenesis has been suspected. Previous our study showed that glycolic acid, one of the most commonly used AHAS increased skin initation dose dependentty aiter treatment for 14 Consecutive days. In the present study. we examined the tumor (anti)promoting ability of glycolic ‘acid on two-stage carcinogenesis test using inbred hairless female mice (15/group) skin tumors either induced by 7,12-dimethylbenz alanthracene (OMBA) as an initiator and glycolic acid (twice ‘2 week) as a promoter, or induced by UVB followed glycolic acid (12.5 mg/cm). Glycolic acid ‘promoted parpiioma incidence and multiplicity initiated by MBA similar to 12—O-telradecanoy! ‘Bhorbol-13-acetate (TPA), however, inhibited UVB-induced parploma formation. The expressions ‘of PCNA, cyclins, cyclin dependent kinase and cyclooxygenase-2 . and the activation of transcription factor NF-KB and AP-1 were concomitantly decreased in glycolic acid treated skin compared to UVB treated skin. Change of these factors by glycolic acid may collectively contribute to during the skin carcinogenesis. IpA4=13] [ 10/19/2000 (Thr) 10:00 = 11:00 / [Hall 8) ] ‘The Roles of ATP and Calcium in Morphology Changes and Cytotoxicity Induced by Benzoquinone in Platelets 164Lee SK°, Chung SM. Lee MY, Chung JH College of Pharmacy, Seoul National University, Seoul 151-742 Korea To understand mechanism of benzoquinoneinduced cytotoxicity, the roles of ATP and calcium in platelet toxicity and morphology changes was investigated. Using scanning electron microscopy. morphological changes to platelets following 1,4-benzoquinone exposure consisted of membrane blebbing at § min which was significantly different from shape changes (pseudopod formation) ‘observed in response to physiological ag (LOH leakage) observed be mins was dependent on extraceliar calcium and associated with increased cytosol . Benzoquinone-induced cytotoxicity was inhibited by calmodulin antagonists, suggesting that Calmodulin could play @ major role in 1,4benzoquinone toxicity via protease activation. These results suggested thet the progression of events for quinone-induced cytotoxicity in platelets to be as follows: quinones deplete intracellular ATP: formation of blebs occurs: calcium homeostasis is disrupted, resulting activation of calmodulin-dependent proteases: ireversible cytotoxicity occurs. [pA4~14] [ 10/19/2000 (Thr) 10:00 ~ 11:00 / {Hall 8) } ‘TCDD induced micronuciel in estrogen receptor positive human breast cancer colls im JW, Han ES, Park MS, Eom MO, Jun HS, Jung HK, Sim WS and Oh HY Genetic Toxicology Division, National institute of Toxicological Research. KFOA, Laboratory of Molecular Biology, Department of Biology, Korea University Bisphenol A (BPA, Cas no. 8005-7), di~2-ethylhexyl phthalte (DEHP, Cas no. 117-81-7), and 2,8,7,8-tetrachlorodibenzodioxin (TCDO, Cas no. 1746-01~6) were well known endocrine disrupting chemicals (EDCs). They showed all negetive results In the Standard genetic toxicology test battery recommended by ICH quid actorial reverse mutation assay. chromosome aberration assay. mouse lymphoma tk+/~ assay and in vivo rodent micronucleus assay (MN). In ‘ur previous study. bisphenol A and di~2-ethylnexyi phthalte induced micronucleus formation in MCF-7 cells (estrogen receptor positive). In this study. to identity the relationship between MN formation and estrogen receptor (ER), TCDD was studied using micronucleus formation in human breast MCF~7 cells (ER positive). We also performed in vito MN to identiy the role of estrogen receptors with TCOD and tamoxifen, inhibitor of estrogen receptor, in MCF-7 cells during micronucleus formation. TCDD induced MN formation in MCF-7 cel's (ER positive) was 6.60 fold higher than that of MCF-10A celis (ER negative). Though TCOO induced MN in MCF-10A cells, the ‘equencies were weak positive, Tamoxifen inhibited TCDD-induced MN formation up to 47.3% in [PA4~15] [ 10/19/2000 (Th) 10:00 ~ 11:00 / {Hall 8) } Potentiation of Agonist-induced Platelet Aggregation by Trivalent Arsenic Lee MY°, Bae ON, Chung JH College of Pharmacy, Seoul National University, Seoul 151-742 Korea Chronic ingestions of arsenic by drinking water have been shown to Induce cardiovascular disease 165,such as blacktoot disease, atherosclerosis and hypertension. but the exact machanism has not bbeen elucidated yet. In order to investigate one of the possible causes toward cardiovascular disease by arsenic, we examined the effects of arsenic on platelets which play an important role in development of cardiovascular disease. Addition of sodium arsenite (Asil) tivalent inorganic arsenic. to rat platelets did not induce either aggregation or cytotoxicity to platelets directly, whereas Asitl treatment potentiated piatelet aggregations induced by various agonists, such as thrombin, collagen, ADP and arachidonic acid in concentration~ and time-dependent manners. ‘Thrombin-induced platelet agoregation was also enhanced by relatively higher concentration of sodium arsenate (AsV) or monomethylarsonic acid (MMA) compared to Asill. Treatment with Asill resulted in a dose-dependent elevation of thrombin-induced serotonin levels trom platelets, while the formation of thromboxane A2 from platelets did not altered significantly. Consistent with these findings. the in vivo studies revealed that ingestion of drinking water containing Ail in mouse increased blood serotonin levels significantly which is indicative of platelet agoregation in vivo. ‘These results suggest that Asill exposure makes platelets more susceptible to agonist-induced ‘agaregation mediated through serotonin secretion from platelets and thus these effects by Asill may contribute to the pathogenesis of cardiovascular disease. (PA4~16] [10/19/2000 (Thx) 10:00 ~ 11:00 / {Hall 8} } Studies on ONA damage by single cell gel electrophoresis and endocrine disrupting activity by transcriptional assay of dibutyl phthalate Kim HT? '-*, chai ¥G?, Ryu scl, "Toxicology Laboratory, Korea Institue of Science and Technology, Seoul, 136-650, Korea, 20epertment of Blochemistry and Molecular Biology, Hanyang Univesity, 425-791, Korea ‘A wide range of phthalates have been produced for use as plasticizers and softeners in many ‘synthetic products. Among phthalate esters, Di-n-butyi-phthalate (08°) may act as, xenoestiogens or antiandragens. Also, OBP was reported to be genotoxic on human mucosa. To ‘clucidate the relationship between endocrine disrupting activity and ONA damage of phthalate esters, DBP was studied by yeast-based steroid hormone receptor gene transcription essay and single cell gel electrophoresis. We have used a yeast-based assay 10 assess the interactions of DBP with the estrogen, endrogen, and progesterone receptors, DBP ranging from 10718 to 10°"! M was active in the estrogen receptor assay. but It did not show the effect on B-galactosidase activity in the progesterone and the androgen receptor assays. AlS0, to determine whether OBP induces DNA strand breakage, single cell gel electrophoresis (comet assay) was performed using mouse lymphoma L5178Y cell ines. The induction of strand breaks by DBP was not significantly Giterent from contol. In these assays, we found that DBP does not induce ONA single strand breakage in the single cell gel electrophoresis and DBP has estrogenic activity in the gene transcription assay of yeast-based steroid hormone receptor. [PA4~17] { 10/19/2000 (The) 10:00 ~ 11:00 / (Hall 8} ) Establishment of assay to screen estrogenic activity of chemicals Kim YW°, sheen YY College of pharmacy, Ewha womans university To establish the rapid and easy-to-perform methods to screen estrogenic activity of many compounds, we determined 5'-ERE-regulated transactivation and cell proliferation in MCF~7 cells by luciferase assay and SAB assay. respectively. MCF-7 stable celis which are stably transfected with phERE-Luc were treated with many chemicals and then luciferase activity were determined, Estradiol (E2) and synthetic estrogen, diethylstylbesterol (DES) were induced luciferase activity in 166dose dependent manner and thelr induced activities were decreased by tamoxiten (Tam) treatment. Phenolic compounds, such @ octyl pheno! (OP), nonyl phenol (NP). biphenol (BP), also induced the luciferase activity in dose dependent manner. Curcumin-derivatives, such as B18, $8123, induced the luciferase activity and Tam treatment decreased SB118- and SB123~ Induced luciferase activities. Other curcumin-derivative, SB100, didn't induce the luciferase activity, but inhibited OP-, NP and BP-induced luciferase activity. Over than 30 flavonoids were tested in this system, and isoflavone. such as biochanin A, daidzein, genistein, showed higher luciferase activity than others. Resveratrol driven trom red wine induced the luciferase activity in dose dependent manner. To determine cell proliferative effect of chemicals, SAE assay was performed. E2 and DES increased the SRB readings 20-30 folds over that of control, and thelr activities were blocked by Tam treatmont. Mary flavonoids were tested in this system. and similar results to luciferase assay wore achieved. These data shows that these methods are valuable tools for screening estrogenic activity of chemicals {PA4~18] { 10/19/2000 (The) 10:00 ~ 11:00 / {Hall 8) 1 Reduced generation of reactive oxygen species and proliferation in human neuroblastoma cells treated with 2,3,7,8 -tetrachlorodibenzo-p-dioxin Kim J °°, Huh K, Jin DO, Park SH, Lee EH, 1Kang YS, Lee SH, and 1Lee YS. Coll. of Pharmacy, Yeungnam Univ... Kyongsan, #712-749, Korea and Dept. of Physiology. Col, (of Med., Kwandong Univ., Kangnung, #210-701, Korea 2,8,7,8-tetrachlorodibenzo-p-dioxin (TCDD) Is one of the most toxic environmental pollutants. Wide range of toxic effects of TCOD have bean known to be mediated through a ligand activated transcription tactor termed ayinydrocarbon receptor (Ahr). which acts in concert with another structurally related protein, the aryihydracarbon nuclear translacator. Despite the enormous reports regarding diverse actions of TCDD, the direct effect on the central nervous system has been largely unknown. In this study we have examined the toxic effects of TCDD on the human brain derived neuroblastoma cells. TCDD significantly suppressed proliferation of SK-N-SH cals. To elucidate the action mechanism, we studied possible involvement of reactive oxygen species and oxidative stress since endogenously generated reactive oxygen species are important growth modulatory signals. TCDO significantly reduced lipid peroxidation and generation of superoxide anion in the cells. The effect was not blocked by the treatment with a-naphthotlavone, @ Ahr antagonist, or 8-methoxypsoralen. a binding inhibitor of activated Ahr to dioxin responsive element indicating that superoxide reducing action of TCDO is independent from its intracellular receptor. TCDD also significantly inhibited the activites of glutathione reductase, glutathione peroxidase. However, TCOD enhanced the actvity of superoxide dismutase, In conjunction with the fact that a Darticulaly risk group may be newbom infants, as it has been shown that TCOD Is very efficiently transferred by lactation. the results suggest that TCDD may distur brain development through Inhibition of neuronal proliferation and generation of endogenous reactive oxygen species, Supported by Korea Food & Orug Administration. {PA4~19] [ 10/19/2000 (The) 10:00 = 11:00 / [Hall 8) J Gene expression profile and estrogenicity of dibutyl phthalate in MCF7 cells using CDNA microarray and E-screening test Kim HT°, Ryu JC Toxicology Laboratory, Korea Institute of Science and Technology, Seoul, 136-650, Korea Various phthalate compounds are used as softeners and plasticizers in a wide range of plastic materials, Since these substances are nat limited to the original products, but enter the 167environment, they have become widespread envionmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. Among phthelate esters. clbutyl phthalate (DBP) is reported to have estrogenic activity. To elucidate estrogenic activity of DBP. it was studied by E-soreen test and cONA microarrays in MCF7 human breast cancer celis. The E-screen test uses ‘estrogen-sensitive human breast MCF7 cells and compares the cell yield achieved after 6 days of Couture in the medium supplemented with 5% charcoal-dextran stripped fetal bovine serum (FBS) with diverse concentrations of 17B-estradiol and OBP. 17B-estradiol of 107° M and DBP of 10-7 M wore active in tho E-Screen test. Based on the established doses, we compared the pattern of ‘gene expression with the cDNA microarray. It showed some of variation in gene expression patterns among MCF7 cells treated with 17B-estradiol and BP. [PA4-20] [10/19/2000 (The} 10:00 ~ 11:00 / {Hell 8} ‘Suppressive Effect of Bisphenol A on the Cytochrome P450 1A1 induction in Hepa-tctc7 Calls kim J¥°, Choi CY, Jeong HG Department of Pharmacy, Chosun University, Kwangiu, Korea Bisphenol A (4.4'=isopropylidenedipnenol) is a monomer in polycarbonate plastics and a constituent of epoxy and polystyrene resins that are used extensively In the food-packaging Industy and it nas been shown to possess estrogenic properties. n the present study, we investigated the effect of bisphenol A on TCOD- inducible P450 1A1 gene expression in mouse hepatoma Hepa-1c1c7 cells. 2.3,7.8-Tetrachlorodibenzo-p-dioxine (TCOD)induced cytochrome 450 1A1-specilic 7-ethoxyresorutin O-deethylase (EROO) activity was markedly reduced in the concomitant treatment of TCDD and bisphenol A in @ dose dependent manner. TCOD~induced P450 1A1 mRNA lavol was also markedly suppressed in the concomitant treatment of TCDD and bisphenot A. Transient transfection assay using dloxin-response element (DRE)=linked luciferase revealed that bisphenol A reduced transformation of the aryl hydrocarbons (An) receptor to a form capable of specifically binding to the ORE sequence in the promoter of the P450 1A1. These Fasults suggest tne down regulation of the P460 1A1 gene expression by bisphenol A in Hena~ Tcte7 cells might be antagonism of the ORE binding potential of nuciear Ah receptor, (As-21] { 10/19/2000 (Thx) 10:00 ~ 11:00 / [Hall 8] 1 MEASUREMENTS OF ESTROGEN LIKE AND DIOXIN LIKE ACTIVITIES IN KOREAN RIVER WATER USING REPORTER GENE SYSTEM. Joung KE®, Chung KH, Sheen YY College of pharmacy, Ewha womans University The endocrine system Is a complex network of glands and hormones that regulates many of the body's functions, including growth, development and maturation, as wel as the way various ‘organs operate. The endocrine glands (the pituitary. thyroid, adrenal, thymus, pancreas, ovaries, ‘and testes) release carefuly-measured amounts of hormones into the bloodstream that act as natural chemical messengers, traveling to different parts of the body in order to control and adjust many life functions. Endocrine disruptor is @ synthetic chemical that when absorbed into the body either mimics ot blocks hotmones and disrupts the Dody’s normal functions. This distuption can happen through altering normal hormone levels, halting or stimulating the production of hormones. of changing the \way hormones travel through the body, thus affecting the functions thet these hormones control Chemicals that are known human endocrine disruptors include diethylstibesterol (the drug DES), loxin, PCBs, DDT. and some other pesticides. Domestic and industrial effivents have been discharged to Kumho Aver, Kum River, Mankyung River and Mino Stieam of Korea, so contaminated with various organic compounds. We have 168examine the estrogenic and dioxin ke activites using pERE-Luc and pOYP1A1-Luc reporter system. Aver water was extracted using combined solid-phase extraction in static adsorption mode with soxhlet extraction. Chemicals adsotbed to the XAD: + resin were recovered 98,24 5.90% by elution with ethyl acetate and methylene chloride (1:9), Kumho River of Korea showed 0.77 DM E€@ in upstrean and 7.7oM EEQ in downstream. Kum River of Korea showed 3.5pM and |.7pM EEQ in upstream and downstream respectively. Mankyung River of Korea showed 611M and 0.41 pM EEG in upstream and downstream respectively. Mino Stream of Korea showed 0.2pM EEQ only in the upstream. All these samples were tested with PCYP1AI-Luc activity and results showed there were more dioxin lke activities in sediments than water from the river. [PA4~22] [ 10/19/2000 (Thr) 10:00 ~ 11:00 / {Hall 8) } No teratogenicity of Phthalates using in vitro battery system Kim SH®,kim $8, Kwon OR, Sohn KH, Kwack SJ, Oh SD, Choi YW, Han SY, Ha KW, Chung SY and Park KL Reproductive and Developmental Toxicology Division, National institute of Toxicological Research, Korea FDA: College of Pharmacy. Kyunghee University, Seoul, Korea Phthalates have been used as plasticizers in polwinyl chloride plastics such as cable coating. flooring, and blood begs. it was generally demonstrated that mary phthalates is a developmental toxicant in rodents, However. in vito teratogenic effects of phthalates are not clearly known. The aim of this study was to investigate the teratogenic potential of phthalates (DEHP, BBP, and O8P) Using in vitro battery system. Short-term in vito battery system (whole embryo culture and limbbud and midbrain cell micromass culture) has been proposed as a preliminary screening method of teratogens.in whole embryo culture. rat embryos at gestation day 3.5 were cultured in rat IC serum for 48 h. Micromass culture of embiyonic imbbud and midbrain colis was performed based on the method of Flint. Alter 5 days of culture, cell praliferation was assassed by neutral red uptake and Coll differentiation was determined by hematoxylin-stained foci area or alcin blue staining, respectively. In whole embryo culture, there were no morphological abnormalities of embiyo at any concentration of ohthalates. However, phthalates tested in our studies decreased growth and development of embryo only at higher concentration. Although in vito battery eystem did not detect the embryotoxicity of ohthalates, these results suggest that phthalates (OEHP, DBP, and BBP) itself ere able to alter normal embryonic growth and development. {PAa4-23] [ 10/19/2000 (Th) 10:00 = 11:00 / [Hal 8) } Comparative evaluation of a 20-day thyrold/pubertal male assay and Hershberger assay for the detection of androgenic/antiandrogenic activity Moon Hus, Kim HS, Shin JH, Kim TS, Kang IH, Suk JH, Kim IY, Lee KM, Hwang, IK and Han SY Endocrine Toxicology Division, National Institute of Toxicological Research, Korea Food and Orug ‘Administration, 5 Nokoun-dong, Eunpyung-ky 122-704. Seoul Several diferent screening and testing methods for the detection of endocrine disruptors (EDCs) have been proposed recently. A rodent Hershberger assay is one of the screening methods recommended by EDSTAC and OECD. A rodent 20-day thyrold/pubertal male assay is also one of alternative methods to replace the Tier | Screening Battery. The purpose of our study is to evaluate ‘comparatively short-Ierm in vivo screening methods to detect substances with _androgenic/antiandrogenic activity. Hershberger assay was performed ullizing Immature Spragui Dawley mele rats castrated at 6 weeks of age. Testosterone (0.4 mg/kg/day) was subcutaneously (s.c.) injected for 10 days. Additionally, a pure androgen antagonist, flutamide (1. 5, and 10 mg/kg/day) wes administered by o'al gavage ater testosterone treatment. In testosterone tieatment group, glans penis (GP). seminal vesicles (SV), ventral prostate (VP), levator ani muscle 109