할로아세틸시코닌 유도체의 합성 및 항암성 평가 : Synthesis and Evaluation of Antitumor Activity

鄭相國,金光洙,송규용,조훈,안병준 충남대학교 약학대학 의약품개발연구소 1998 藥學論文集 Vol.14 No.-

The secondary hydroxy group at side chain of shikonin structure was selectively acylated with various haloacetic acids in presence of dicyclohexylcarbodiimide and 4-dimethylaminopyridine to produce haloacetylshikonin derivatives. The cytotoxicity of monohaloacetylshikonin derivatives against L1210 cells increased in the following order: monochloroacetylshikonin (ED_50, 0.142 ㎍/㎖) nonobromoacetylshikonin (ED_50, 0.158㎍/㎖)>monoiodoacetylshikonin (ED_50, 0.173 ㎍/㎖). Introduction of larger halogen atoms decreased the cytotoxic activity, presumably due to steric hinderance. The cytotoxicity of chloroacetylshikonin derivatives was dependent on the number of chlorine atom, thus increasing in the following order : trichloroacetylshikonin (0.032 ㎍/㎖)>dichloroacetylshikonin (0.059 ㎍/㎖)> monochloroacetylshikonin (ED_50, 0.142 ㎍/㎖). Thus, the electron-withdrawing effect seems to be important for the cytotoxicity of chloracetylshikonin derivatives. Consistent with the above, dichloracetylshikonin (T/C, 182%) and trifluoroacetylshikonin (195%) showed higher T/C values than monochloroacetyl-(T/C, 122%), monobromoacetyl-(T/C, 154%) and monoiodoacetylshikonin (T/C, 117%) derivatives. Haloacetylshikonin derivatives showing lower cytotoxic activities against L1210 cells exhibited lower T/C values. It seems that there is a relationship between the cytotoxicity of haloacetylshikonin and their antitumor activity.

GRIM-19 Expression and Function in Human Gliomas

Jin, Yong-Hao,Jung, Shin,Jin, Shu-Guang,Jung, Tae-Young,Moon, Kyung-Sub,Kim, In-Young The Korean Neurosurgical Society 2010 Journal of Korean neurosurgical society Vol.48 No.1

Objective : We determined whether the expression of GRIM-19 is correlated with pathologic types and malignant grades in gliomas, and determined the function of GRIM-19 in human gliomas. Methods : Tumor tissues were isolated and frozen at $-80^{\circ}C$ just after surgery. The tissues consisted of normal brain tissue (4), astrocytomas (2), anaplastic astrocytomas (2), oligodendrogliomas (13), anaplastic oligodendrogliomas (11), and glioblastomas (16). To profile tumor-related genes, we applied RNA differential display using a $Genefishing^{TM}$ DEG kit, and validated the tumor-related genes by reverse transcription polymerase chain reaction (RT-PCR). A human glioblastoma cell line (U343MG-A) was used for the GRIM-19 functional studies. The morphologic and cytoskeletal changes were examined via light and confocal microscopy. The migratory and invasive abilities were investigated by the simple scratch technique and Matrigel assay. The antiproliferative activity was determined by thiazolyl blue Tetrazolium bromide (MTT) assay and FACS analysis. Results : Based on RT-PCR analysis, the expression of GRIM-19 was higher in astrocytic tumors than oligodendroglial tumors. The expression of GRIM-19 was higher in high-grade tumors than low-grade tumors or normal brain tissue; glioblastomas showed the highest expression. After transfection of GRIM-19 into U343MG-A, the morphology of the sense-transfection cells became larger and more spindly. The antisensetransfection cells became smaller and rounder compared with wild type U343MG-A. The MTT assay showed that the sense-transfection cells were more sensitive to the combination of interferon-$\beta$ and retinoic acid than U343MG-A cells or antisense-transfection cells; the antiproliferative activity was related to apoptosis. Conclusion : GRIM-19 may be one of the gene profiles which regulate cell death via apoptosis in human gliomas.

Dual roles of a variable number of tandem repeat polymorphism in the TERT gene in lung cancer

Jin, Guang,Yoo, Seung Soo,Cho, Sukki,Jeon, Hyo-Sung,Lee, Won-Kee,Kang, Hyo-Gyoung,Choi, Yi Young,Choi, Jin Eun,Cha, Sung-Ick,Lee, Eung Bae,Kim, Chang Ho,Jung, Tae Hoon,Kim, Young Tae,Park, Jae Yong Wiley (Blackwell Publishing) 2011 CANCER SCIENCE Vol.102 No.1

Genomic structure of the Spodoptera litura multicapsid nucleopolyhedrovirus

Yong Wang,Jae Young Choi,Hee Jin Shim,Jong Yul Roh,Hong Guang,Qin Liu,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05

The complete genomic nucleotide sequence of the Spodoptera litura multicapsid nucleopolyhedrovirus (SlMNPV) isolated in Korea, SlMNPV-K1, was determined. It was 137,435 bp long, with a 55.4 % A+T content and contained 132 putative open reading frames (ORFs) of 150 nucleotides or larger that showed minimal overlap. The 132 putative ORFs covered 87.7% of the genome. Among these, 131 ORFs were are homologous to genes identified in previously reported SlMNPV genome which consisted 139,342 bp and contained 141 putative ORFs. However, arrangement of some ORFs were somewhat different from each other. Even though the SlMNPV-K1 genome is smaller than that of previously reported SlMNPV genome and had lesser predicted ORFs, the main functional genes were all conserved. When the phylogenic relationship was analyzed using the nucleotide sequence of polyhedrin gene, SlMNPV-K1 was most closely related to Lymantria dispar multicapsid nucleopolyhedrovirus (LdMNPV) which were belonged to Group Ⅱ nucleopolyhedrovirus.

Apoptosis Inducing Effects of 6-Methoxydihydrosanguinarine in HT29 Colon Carcinoma Cells

Yong-Jin Lee,Hu-Quan Yin,Young-Ho Kim,Guang-Yong Li,Byung-Hoon Lee 대한약학회 2004 Archives of Pharmacal Research Vol.27 No.12

6-Methoxydihydrosanguinarine (6ME), a benzophenanthridine alkaloid derived from the methanol extracts of Hylomecon hylomeconoides, showed a dose-dependent effect at 1-10 μM on causing apoptotic cell death in HT29 colon carcinoma cells (IC50 = 5.0 ± 0.2 μM). Treatment of HT-29 cells with 6ME resulted in the formation of internucleosomal DNA fragmentation. Treatment of the cells with 6ME caused activation of caspase-3, -8 and 9 protease and subsequent proteolytic cleavage of poly(ADP-ribose)polymerase. 6ME increased the expression of p53 and Bax and decreased the expression of Bid. These results indicate that p53 and proapoptotic Bcl-2 family proteins might participate in the antiproliferative activity of 6ME in HT29 cells.

Development of Biomarkers for Cadmium Exposure Using Toxicogenomic Technique

Guang-Yong Li,Mi-Ock Lee,Yong-Jin Lee,Chang-Kiu Moon,Byung-Hoon Lee 대한약학회 2004 大韓藥學會 總會 및 學術大會 Vol.2004 No.1

Synthesis and Evaluation of Antitumor Activity

Jin, Guang-Zhu,Song, Gyu-Yong,Zheng, Xiang-Guo,Kim, Yong,Sok, Dai-Eun,Ahn, Byung-Zun The Pharmaceutical Society of Korea 1998 Archives of Pharmacal Research Vol.21 No.2

Fourty eight derivatives of 2-(1-oxyalkyl)-1,4-dioxy-9,10-anthraquinone were synthesized, and their antitumor activity was evaluated. On the whole, 2-(1-hydroxyalkyl)-1,4-dihydroxy-9,10-anthraquinones (DHAQ=1,4-dihydroxy-9,10-anthraquinone) showed stronger cytotoxic activity against L1210 cells than 2-(l-hydroxyalkyl)-1,4-dimethoxy-9,10-anthraquinones(DMAQ =1,4-dimethoxy-9,10-anthraquinone), implying that free hydroxy groups at C-1 and C-4 of the anthraquinone structure are necessary for the cytotoxic activity. The bioactivity of 2-(lhydroxyalkyl)-DHAQ derivatives differed according to the size of alkyl group at C-1;while the elongation of alkyl group over 7 carbon atoms failed to enhance the bioactivity, the derivatives possessing alkyl moiety of 1-6 carbon atoms showed an increase in the cytotoxicity and the antitumor activity in Sarcoma-180; 2-hydroxymethyl-DHAQ ($ED_{50}$, $15\mu\textrm{g}$/ml; T/C, 125%), 2-(1 -hydroxyethyl)-DHAQ($1.9{\mu}g/ml;139.2%)$;, 2-(1-hydroxypropyl)-DHAQ ($7.2{\mu}g$/ml; 135.1%), 2-(1-hydroxybutyl)-DHAQ ($10.2{\mu}g/ml; 125.3%)$, 2-(1-hydroxypentyl)-DHAQ ($23.7{\mu}g/ml; 110.1%$). and 2-(1-hydroxyhexyl)-DHAQ ($58{\mu}g/ml;108%$). Next, 2-(1-Hydroxyalkyl)-DHAQ derivatives were acetylated to produce 2-(1-acetoxyalkyl)-DHAQ analogues. Although the acetylation somewhat enhanced the cytotoxicity, but not the antitumor action. In addition, the presence of phenyl group at $C-1^{l}$ enhanced the cytotoxicity and the T/C value, compared to alkyl groups of same size; 2-(1-hydroxy-1-phenyl)-DHAQ ($ED_{50}$, $5.6{\mu}g$, T/C, 137%).

The Effect of Hyaluronic Acid on the Invasiveness of Malignant Glioma Cells : Comparison of Invasion Potential at Hyaluronic Acid Hydrogel and Matrigel

Jin, Shu-Guang,Jeong, Young-Il,Jung, Shin,Ryu, Hyang-Hwa,Jin, Yong-Hao,Kim, In-Young The Korean Neurosurgical Society 2009 Journal of Korean neurosurgical society Vol.46 No.5

Objective : Hyaluronidase (HAse), a degrading enzyme of hyaluronic acid (HA), is highly expressed in patients with malignant glioma. The purpose of this study was to verify whether HAse is related to the invasion of glioma cells. We also investigated if glioma cells with higher mobility in 2-dimensioal (2-D) method have also higher mobility at 3-dimensional (3-D) environment. Methods : Malignant glioma cell lines (U87MG, U251MG, U343MG-A, and U373MG) were used, and their HAse expressions were evaluated by HA zymography. The migration ability was evaluated by simple scratch technique. The invasiveness of each cell lines was evaluated by Matrigel invasion assay and HA hydrogel invasion assay. In HA hydrogel invasion assay, colonies larger than $150\;{\mu}m$ were regarded as positive ones and counted. Statistical analysis of migration ability and invasion properties of each cell lines was performed using t-test. Results : In scratch test to examine migration ability of each cell lines, U87MG cells were most motile than others, and U343MG-A least motile. The HAse was expressed in U251MG and U343MG-A cell lines. However, U87MG and U373MG cell lines did not express HAse activity. In Matrigel invasion assay, the cell lines expressing HAse (U251MG and U343MG-A) were more invasive in the presence of HA than HAse deficient cell lines (U87MG and U373MG). In HA hydrogel invasion assay, the HAse-expressing cell lines formed colonies more invasively than HAse-deficient ones. Conclusion : Malignant Glioma cells expressing HAse were more invasive than HAse-deficient ones in 3-dimensional environment. Therefore, it might be suggested that invasion of malignant gliomas is suppressed by inhibition of HAse expression or HA secretion. Additionally, the ability of 2-D migration and 3-D invasion might not be always coincident to each other in malignant glioma cells.

Construction of modified Bacillus thuringiensis cry1AC genes based on cry1-5 genes through multi site-directed mutagenesis

Hong Guang Xu,Jong Yul Roh,Jae Young Choi,Hee Jin Shim,Yong Wang,Qin Liu,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05

Bt crystal proteins, encoded by cry genes, are a group of insecticidal proteins unique in the Gram-positive and spore-forming bacterium, Bacillus thuringiensis. These cry genes are widely applied as one of the most successful candidates for constructing transgenic plants resistant to pest insects. In our previous report, we found Cry1-5 had high insecticidal activity against Spodoptera larvae although its amino acid sequences showed high similarity (97.9%) to those of Cry1Ab which had low activity. In comparison with Cry1Ac, Cry1-5 had 12 different residues in domain Ⅰ and domain Ⅱ, and we focused on domain Ⅰand domain Ⅱ regions and designed 10 mutagenic primers to change 12 residues. Through multi site-directed mutagenesis, we mutated the modified cry1Ac gene by plant codon usage in pOBⅠ-Mod-cry1Ac based on cry1-5 and constructed 63 various mutant cry genes. In the further study, we will express those mutant proteins as a fusion form with polyhedrin using baculovirus expression system and subsequently do bioassay to Spodoptera larvae.

Apoptosis Inducing Effects of 6-Methoxydihydrosanguinarine in HT29 Colon Carcinoma Cells

Lee, Yong-Jin,Yin, Hu-Quan,Kim, Young-Ho,Li, Guang-Yong,Lee , Byung-Hoon The Pharmaceutical Society of Korea 2004 Archives of Pharmacal Research Vol.27 No.12

6-Methoxydihydrosanguinarine (6ME), a benzophenanthridine alkaloid derived from the methanol extracts of Hylomecon hylomeconoides, showed a dose-dependent effect at 1-10 ${\mu}M$ on causing apoptotic cell death in HT29 colon carcinoma cells $(IC_{50} = 5.0{\pm}0.2 {\mu}M)$. Treatment of HT-29 cells with 6ME resulted in the formation of internucleosomal DNA fragmentation. Treatment of the cells with 6ME caused activation of caspase-3, -8 and 9 protease and subsequent proteolytic cleavage of poly(ADP-ribose)polymerase. 6ME increased the expression of p53 and Bax and decreased the expression of Bid. These results indicate that p53 and proapoptotic Bcl-2 family proteins might participate in the antiproliferative activity of 6ME in HT29 cells.