11:10-12:00, Rm 103

Cancer biologyB-17-50Thymoquinone (TQ) Modulates Cyclooxygenase-2 Expression throughPI3kinase and p38 Kinase Pathway in Human Breast Cancer Cell Line,MDA-MB-231Seon Mi-Yu and Song-Ja KimDepartment of Biological Sciences, Kongju National University, Gongju 314-701, KoreaThymoquinone (TQ), a drug extracted from Nigella sativa of black seed, has shown toexhibit anti-inflammatory, anti-oxidant and anti-neoplastic effects in various cancer cells.The effect of TQ on cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2)production in MDA-MB-231 remains poorly understood. For this reason, we haveinvestigated the effect of TQ on expression of COX-2 in MDA-MB-231. TQ significantlyinduced COX-2 expression and PGE2 production in a dose- and -time dependentmanner as determined by Western blot analysis and PGE2 assay. Also, TQ increasedactivation of Akt and p38kinase. Inhibition of p38kinase with SB203580 and PI3Kinasewith LY294002 was abolished TQ caused COX-2 expression and PGE2 production.Taken together, these results collectively demonstrate that TQ effectively modulatesCOX-2 expression via PI3Kinase and p38kinase pathway in MDA-MB-231. Additionally,this pathway may provide a pivotal understanding for therapeutic inhibition of cancer.B-17-51Identification of autoantibody against fatty acid synthase (FASN) inhepatocellular carcinoma (HCC) mouse model and its application todiagnosis of HCCChang-Kyu Heo 1, 7 , Mi-Kyung Woo 1 , Dae-Yeul Yu 2 , Ju Yeon Lee 3 , Jong Shin Yoo 3 ,Hyang Sook Yoo 1 , Jeong Heon Ko 1 , Jin-Man Kim⁴, Jong Young Choi 5 , Sang GiPaik 7 , Eun-Wie Cho 1¹Daejeon-KRIBB-FHCRC Research Cooperation Center, Korea Research Institute of Bioscience andBiotechnology (KRIBB), Daejeon 305-860, ²Department of Human Genomics, KRIBB, ³Division ofProteome Research, Korea Basic Science Institute, Daejeon 305-333, ⁴Department of Pathology, Collegeof Medicine, Chungnam National University, Daejeon 301-131, 5 Department of Internal Medicine,College of Medicine, the Catholic University of Korea, Seoul 137-040, 7 Department of Biology, School ofBiosciences and Biotechnology, Chungnam National University, Daejeon 305-764, KoreaAutoantibodies, which are generated by immune system recognizing the presence of theabnormal tumor-associated antigens, are promising biomarkers for early detection oftumors. Recently, we established a B cell hybridoma pool derived from H-ras12Vtransgenic mouse, a typical hepatocellular carcinoma model, as a source of tumorassociatedautoantibodies without using any extracellular antigens and have characterizedthe specific target antigens against them. K1 autoantibody, one of them, was investigatedin this study and its target antigen was identified by mass spectrometric analysis as fattyacid synthase (FASN), an important oncogenic protein. Moreover, a specific mimotopeagainst K1 autoantibody was screened from the cyclic random hepta-peptide phage libraryand, using it as a coating antigen for ELISA, we could distinguish patients withhepatocellular carcinoma (HCC) vs. normal subjects with 96.55% sensitivity and 100%specificity. These results imply that anti-FASN autoantibody is induced in patients withHCC and detection of anti-FASN autoantibody can be used for the diagnosis of HCC.B-17-52Interleukin-5-induced MMP-9 expression is mediated by the ERK1/2,AP-1 and NF-κB pathway in 5637 bladder cancer cellsEo-Jin Lee¹ , ², Wun-Jae Kim¹ , ⁴, Se-Jung Lee¹ , ³, Sung-Seok Park¹ , ²and Sung-Kwon Moon¹ , ² , *¹Personalized Tumor Engineering Research Center (PT-ERC), Chung-Buk National University,²Department of Biotechnology, Chung-ju National University, ³Department of Food Science &Technology, Chung-Ang University, ⁴Department of Urology, Chungbuk National UniversityCollege of Medicine, *Corresponding author : Sung-Kwon MoonHuman IL-5 plays an important role in proliferation and differentiation of humaneosinophils. In this study, the role and importance of the signaling pathway in thetranscriptional regulation of MMP-9 in human bladder cancer 5637 cells was examined.Gelatin zymography and immunoblot analysis data demonstrated that IL-5 increased theexpression of MMP-9 in the cultured 5637 bladder cancer cells. Transcription factors NFκBbinding site (-601) and AP-1 binding site (-82) were identified as the cis-elements forIL-5 activation, as determined by gel shift assay. In addition, IL-5 treatment stimulatedphosphorylation of ERK1/2 in 5637 cells. Collectively, our results demonstrate that IL-5stimulates MMP-9 expression via the ERK1/2 signaling pathway in bladder cancer 5637cells.B-17-53Identification of autoantibody against cytokeratin 8/18 complex in H-ras12V tumor model mouse and its application to diagnosis of humanbreast cancerChang-Kyu Heo 1 , Hae-Min Hwang 1 , Dae-Yeul Yu 2 , Ju Yeon Lee 3 , Jong-Shin Yoo 3 ,Hyang Sook Yoo 1 , Jeong Heon Ko 1 , Jin-Man Kim⁴, Sejeong Oh 5 and Eun-Wie Cho 1¹Daejeon-KRIBB-FHCRC Research Cooperation Center, Korea Research Institute of Bioscience andBiotechnology (KRIBB), ²Department of Human Genomics, KRIBB, Daejeon 305-860, ³Divisionof Proteome Research, Korea Basic Science Institute, Daejeon 305-333, ⁴Department of Pathology,College of Medicine, Chungnam National University, Daejeon 301-131, 5 Department of Surgery,College of Medicine, The Catholic University of Korea, Inchon 403-720, KoreaAutoantibodies, which are generated by immune system recognizing the presence of theabnormal tumor-associated antigens, are promising biomarkers for the early detection oftumors. Recently, we established hundreds of B cell hybridomas from H-ras12V transgenicmouse as a source of tumor-associated autoantibodies without using any extracellularantigens and have characterized specific target antigens against them. TAB-K94monoclonal antibody, one of tumor-associated autoantibodies obtained from H-ras12Vtransgenic mouse, was investigated in this study and its target antigen was identified ascytokeratin 8/18(CK8/18) complex, an intermediate filament(IF) protein complex ofepithelial cells which is involved in cell motility and cancer progression. To establish amethod for the detection of autoantibodies against CK8/18 complex in tumor patients sera,a specific mimotope against TAB-K94 autoantibody was screened from the cyclic randomhepta-peptide phage library and, by enzyme-linked immunosorbent assay(ELISA) using itas a binder of anti-CK8/18 autoantibodies, we could distinguish breast cancer patients vs.normal subjects with 50.0% sensitivity and 82.6% specificity. These results imply thatdetection of anti-CK8/18 autoantibody would be useful for the diagnosis of breast cancer.B-17-54Development of a novel validation method with high sensitivity andmultiplexity based on DNA-tagged antibodies for cancer biomarkerdiscoveryChangHee Cho, Yong-Sam Kim, Hyang-Sook Yoo and Jeong Heon KoDaejeon-KRIBB-FHCRC Research Cooperation Center, KRIBB, Daejeon 305-806, KoreaThe development of cancer biomarker is of a great promise to conquer cancer since itleads to an early detection and provide an opportunity for better cancer treatment.Validation is a time-consuming step for biomarker developments requiring investigation ofbiomarker candidates using thousands or more of biosamples, which is why a sensitive,multiplexing validation method is necessary. To address this challenge, we aredeveloping as antibody-based validation method, in which antibody with oxidized N-glycans was covalently linked to an identifiable DNA tag for use of the template in thetranscription by T7 polymerase. The transcripts will be quantified and used for anindicator of the amounts of the antigen bound to the DNA-tagged antibody. This strategywill be multiplexed for an establishment of a novel validation method.B-17-55Loss of p21Sdi1 expression accounts for proliferation of senescent cellsafter DNA strand break along with increased miR-93 expression andreduced p53 binding to p21Sdi1 promoterOk Ran Choi and In Kyoung LimDepartment of Biochemistry and Molecular Biology, Ajou University School of Medicine, Suwon443-721, KoreaTo answer the question what is a critical event for higher incidence of tumor developmentin old than young individuals, primary culture of human diploid fibroblasts were employedand DNA damage was induced by doxorubicin or X-ray irradiation. Old cells differentiallyresponded to DNA double strand breaks; Loss of p21sdi1 expression and poly(ADPribose)-polymeraseinduction in spite of p53S15 activation, and [3H]-thymidine and BrdUincorporations, suggesting cell cycle progression. The phenomena were confirmed byother tissue fibroblasts obtained from different donor ages. Reduced p21sdi1 expressionwas accompanied with miR-93 expression and failure of p53 binding to p21 promoter insenescent cells after DNA damage. Therefore, loss of p21 expression in senescent cellsafter DNA damage may account for in vivo carcinogenic process in aged tissue withoutrepair arrest.176 Korean Society for Biochemistry and Molecular Biology