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Yu Jihn Kwon,So Young Chung,Eun Joo Koo,Ji Eun Park,Dong Hyuk Seo,Yo A Lee,Yu Young Jung,Hee Eun Min,Mi Ran Kim,Eungui Kang,Jeongyun Cho,Seong Soo Park,Sun Ok Choi,Chul Joo Lim 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07
Genetically modified (GM) papaya (Carica papaya L.) line 55-1 (55-1), which is resistant to papaya ringspot virus infection, has been marketed internationally. Many countries such as the European Union, Japan, and Korea have a mandatory safety assessment, approval and labeling regulations for GM foods. Thus, there is a need for specific methods for detecting 55-1. In this study, we established a real-time PCR detection method applicable to 55-1 for a variety of papaya products. The limit of detection was possible for fresh papaya fruit up to dilutions of 0.005% and 0.01% (weight per weight [w/w]) for homozygous SunUp and heterozygous Rainbow cultivars, respectively, in non-GM papaya. The 55-1 event-specific detection method observed parallelism (r2>0.99) between the concentration of line 55-1 cultivars and Ct values obtained in amplification plots at concentrations of 0.005-10% for SunUp DNA and 0.01-10% for Rainbow DNA. The method was applicable to the qualitative detection in various types of processed products (cocktail fruit, dried fruit, juice, etc.) containing papaya as a main ingredient. Monitoring papaya products for the presence of GM papaya were demonstrated using a P35S and T-nos real-time PCR detection method but no amplification signals were detected.
( Mi Jin Lee ),( Goung Ran Yu ),( Hua Lee ),( Jun Zhang ),( Yun A Kim ),( Dae Ghon Kim ) 대한내과학회 2014 대한내과학회 추계학술대회 Vol.2014 No.1
Background: NM23 is a family of structurally and functionally conserved proteins known as nucleoside diphosphate kinases (NDPK). NM23-H1, NM23-H2 and NM23- LV are expressed abundantly in HCC. NM23-H2 is a basic protein recently identifi ed as the human PuF factor, which is a transcriptional activator of the c-Myc proto-oncogene. Although the NM23 protein is implicated as a metastasis suppressor, the role of NM23 appears to be less understood. Thus, the aim of this study is to examine functional role and mechanism of NM23 involved tumorigenesis in HCC. Methods: We examined the NM23-H1, H2 and LV mRNA expression in HCC by Realtime- PCR analysis and NM23-H1, H2 and LV protein expression in HCC by immunoblot and immunohistochemistry. Focus formation and anchorage-independent growth were examined in stable cell lines expressing NM23 using soft agar. Using overexpression of NM23 by adenoviral system, the molecular mechanism of NM23 mediated tumor cell growth was assessed in experimental cell culture and in vivo animal model. Results: The level of NM23 expression in tumor tissues and the surrounding matrix appeared to be independent of etiology and tumor differentiation. Ectopic expression of NM23-H2 in NIH3T3 fi broblasts and HLK3 hepatocytes enhanced focus formation, and allowed anchorage-independent growth. Overexpression of NM23 results in increased tumor cell proliferation, which is relevant to activation of AKT and extracellular signal-regulated kinase (ERK)1/2 phosphorylation. The NIH3T3 fi broblasts and HLK3 hepatocytes stably expressing NM23-H2 produced tumors in athymic mice. Lentiviral delivery of NM23-H2 shRNA inhibited tumor growth of xenotransplanted tumors produced from HLK3 cells stably expressing NM23-H2. Conclusions: These results indicate that NM23 may be pro-oncogenic and involved in hepatocarcinogenesis through AKT/ERK signaling pathway. Therefore, this pathway may be an useful target for HCC treatment.
Basic, HCC basic : O-034 ; Pro-oncogenic NM23-H2 modulates MDM2 expression in hepatocarcinogenesis
( Mi Jin Lee ),( Goung Ran Yu ),( Hua Lee ),( Sang Wook Kim ),( Pei Pei Hao ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1
Background: NM23 is a family of structurally and functionally conserved proteins known as nucleoside diphosphate kinases (NDPK). NM23-H1 and NM23-H2 are expressed abundantly in HCC. NM23-H2 is a basic protein recently identified as the human PuF factor, which is a transcriptional activator of the c-Myc proto-oncogene. Although the NM23-H1 protein is implicated as a metastasis suppressor, the role of NM23-H2 appears to be less understood. Thus, the aim of this study is to examine functional role and mechanism of NM23-H2 involved tumorigenesis in HCC. Methods: We examined the NM23-H1, H2 and LV mRNA expression in HCC by Realtime-PCR analysis and NM23-H2 protein expression in HCC by immunoblot and immunohistochemistry. Focus formation and anchorage-independent growth were examined in stable cell lines expressing NM23-H2 using soft agar. Using overexpression of NM23-H2 by adenoviral system, the molecular mechanism of NM23-H2 mediated tumor cell growth was assessed in experimental cell culture and in vivo animal model. Results: The level of NM23-H2 expression in tumor tissues and the surrounding matrix appeared to be independent of etiology and tumor differentiation. Ectopic expression of NM23-H2 in NIH3T3 fibroblasts and HLK3 hepatocytes enhanced focus formation, and allowed anchorage-independent growth. Ectopic expression of NM23-H2 induced MDM2 expression. However, MDM2 mRNA and promoter activity was not changed by ectopic expression of NM23-H2, but NM-23H2 interacted with MDM2. The NIH3T3 fibroblasts and HLK3 hepatocytes stably expressing NM23-H2 produced tumors in athymic mice. Lentiviral delivery of NM23-H2 shRNA inhibited tumor growth of xenotransplanted tumors produced from HLK3 cells stably expressing NM23-H2. Conclusions: These results indicate that NM23-H2 may be pro-oncogenic and regulate MDM2 expression in hepatocarcinogenesis. Therefore, this pathway may be an useful target for HCC treatment.
Antioxidant Activity of Limonene Perilla and Perilla Cultivar Using Different Solvents
Mi Ran Jeon(전미란),Ji Hye Yoo(유지혜),Jae Hoo Choi(최재후),Chang Heum Kim(김창흠),Byeong Ju Kang(강병주),Jae Geun Lee(이재근),Nam Jun Kim(김남준),Su Jin Yang(양수진),Jung Dae Lim(임정대),Seon Kang Choi(최선강),Chang Yeon Yu(유창연) 한국약용작물학회 2016 한국약용작물학회 학술대회논문집 Vol.2016 No.1
참억새로부터 분리된 S-Adenosylmethionine Synthetase (SAMS) 유전자를 이용한 담배 형질전환체계 확립
Mi Ran Jeon,Eun Soo Seong,Ji Hye Yoo,Jae Hoo Choi,Chang Heum Kim,Byeong Ju Kang,Kweon Heo,Chang Yeon Yu 한국약용작물학회 2016 한국약용작물학술대회 발표집 Vol.2016 No.10
Background : Temperature is major factor for growth plant. Recently, because of global warming, abnormal temperature included drought, deluge, sudden temperature change and heavy snow damaged crops in the world. In Korea, crops have been sensitive to low temperature on early growth stage, e.g. fruit tree and ginseng, were damaged owing to sudden heavy snow and cold on Spring. Therefore, recently interest in cold resistance crops were increased in demand rapidly. This study was performed to establish transgenic Nicotiana benthamiana by transforming cold resistant gene related to cold tolerance S-adenosylmethionine synthetase (SAMS) isolated from Miscanthus sinensis. Methods and Results : Total RNA was extracted from leaves of M. sinensis using Trizol assay and isolated MsSAMS. Isolated MsSAMS was insert into SacⅠ- XbaⅠ sites of pMBP1 vector. The vector was transformed to Agrobacterium tumefaciens strain LBA4404 by DH5α. A. tumefaciens with binary plasmid were selected at YEP medium supplemented with kanamycin. Cut leaves of tobacco were co-cultured with selected A. tumefaciens. Co-cultured leaves was grown on regeneration medium for a month at dark condition, and transferred to at light condition. Regeneration shoot from callus were excised and transferred to root-induction medium. Approximately, 58% of leaves explant produced callus. Nearly, 30% of callus had shoot and approximately, 94% of shoots were rooted in root-induction medium. Conclusion : We established an efficient transformation system of N. benthamiana transformed by using MsSAMS gene related to cold tolerance isolated from M. sinensis. We may use the produced transgenic plants to prevent damages carried by cold.