• 대한예방한의학회지
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• 제5권1호
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• pp.144-150
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• 2001

The cytotoxic activity of Cratalariae sessiliflorae on cultured NIH 3T3 fibroblasts and human oral epithelioid carcinoma cells (KB) were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) colorimetric method These fractions of methanol extract of Cratalariae sessiliflorae showed inhibitory effect in vitro in the milligram range against KB cell lines. In general, the antitumor activities of these fractions were does-dependent over the milligram range. The comparison of IC50 values of these fractions in tumor cell lines showed that their susceptibility to these fractions decrease in the following order: Fr. 4> Fr. 6> Fr. 10> Fr. 2> Fr. 11> Fr. 3> Fr. 8> Fr. 7> Fr. 9> Fr. 1> Fr. 5 by the MTT assay. These fractions were tested for their cytotoxic effects on NIH 3T3 fibroblasts using MTT assay. They exhibited potent cytotoxic activities in vitro in the milligram range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these fractions were does-dependent over the milligram range. The comparison of CD50 values of these fractions in NIH 313 fibroblasts shows that their susceptibility to these fractions in decrease the following order: Fr. 10> Fr. 9> Fr. 2 = Fr. 4> Fr. 8> Fr. 11> Fr. 1 = Fr. 7> Fr. 3> Fr. 5 = Fr. 6 by the MTT assay. These results suggests that fraction 5 has the most growth - inhibitory activity against KB cell lines.

• BMB Reports
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• 제31권3호
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• pp.269-273
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• 1998

The objective of this study was to evaluate the reversal of multi drug resistance of human cell lines by specific inhibitors of ${\alpha}-glycosidase$ and mannosidases that had been reported to be involved in N-linked oligosaccharide processing of glycoproteins. N-methyldeoxynojirimycin, I-deoxynojirimycin, and castanospermine, which were known to be potent inhibitors of both ${\alpha}-glycosidase$ I and II, showed no activity against the multidrug resistant phenotype of the cell lines of SNU1DOX, KB-V1, and MCF-7/ADR. In contrast, I-deoxymannojirimycin, an inhibitor of mannosidase I, resulted in a slight reversal for the vinblastine resistance of the KB-V1 cell line, but did not show any activity toward the other cell lines. Parallel experiments with tunicamycin, an inhibitor of N-linked glycosylation, also resulted in no significant changes in multidrug resistant (MDR) phenotype of the cell lines tested in this work. These observations suggest that the unglycosylation of P-glycoprotein associated with the inhibitor treatments might not be correlated with the reversal of multidrug resistance of the cell lines tested in this study.

• 한국자원식물학회지
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• 제30권6호
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• pp.679-685
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• 2017

Stachys affinis tubers are known for its high content of stachyose and eaten as an edible vegetable. In this work, we assessed on the anti-inflammatory and anti-proliferation activity of a various type of extracts derived from S. affinis tubers. The n-hexane and dichloromethane fractions were showed the high cytotoxicity on the cell lines including RAW264.7 macrophages, HEK293 human kidney cell, A549 human lung cancer cell, KB human oral cancer cell, and a PC-3 human prostate cancer cell. N-butanol and water fractions were not exhibited cytotoxicity on the tested cancer cells, limited in anti-inflammatory and anti-cancer activities. Nevertheless, the ethyl acetate fraction showed little harm to RAW264.7 cells but inhibited the production of nitric oxide (NO) and prostaglandin E2 (PGE2) significantly. In addition, it arrests the cell growth in A549, KB, and PC-3 cell while little cytotoxicity on HEK293 cells. Consequently, these results supported that the ethyl acetate fraction of S. affinis tubers could be a potential anti-inflammatory and anti-cancer ingredient.

• 생명과학회지
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• 제13권4호
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• pp.522-528
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• 2003

본 연구는 출아형 효모 Saccharomyces cerevisiae에서 자외선의 상해 시 이를 정상으로 회복시키는 절제회복 (excision repair) 유전자로 알려진 RADA4의 특성 규명을 위하여 균류 Coprinus cinereus에서 이와 유사한 유전자를 분리하였다. RAD4 유사 유전자를 분리하기 위하여 균류 C. cinereus의 염색체 DNA를 전기영동하여 분리한 다음 효모 RAD4 DNA를 probe로하여 이와 hybridization하였다. 이 결과 RAD4 유사 유전자는 3.2 kb의 insert DNA를 갖고 있었다. 또한 Southern hybridization으로 이 유사 유전자는 fungus C. cinereus의 염색체에 존재함을 확인하였다. 분리한 RAD4 유사 유전자의 전사체 크기는 2.5 kb 였으며, 자외선의 상해 시 전혀 'inducibility가 없음을 Northern hybridization으로 확인하였다. 또한 유사유전자 부분을 삭제하였을 때 이 부분이 없는 세포는 전혀 생존을 못하였다. 이 결과 분리한 RAD4 유사유전자는 세포의 생존에 관여함을 알 수 있었다.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.207-212
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• 2012

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

• Journal of Microbiology
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• 제37권2호
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• pp.86-89
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• 1999

An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.

• 약학회지
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• 제42권5호
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• pp.507-512
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• 1998

In the course of developing novel antitumor intercalating agents. We synthesized 4-carbamoyloxymethyl-l-azaanthraquinones 7-12, incorporating the latent alkylating functi onality. These compounds were designed to explore the effect of substituent on the nitrogen of carbamate. The target compounds were prepared by hetero Diels-Alder reaction as a key step followed by functionalization of benzylic methyl to the desired substituents. Growth inhibitory studies of the azaanthraquinones were conducted in vitro against human cancer cell lines (SNU-354; liver and MCF7; breast) and human epidermoid carcinoma cells that are sensitive (KB-3-1) and multidrug-resistant (KB-V-1). The compounds were less potent than doxorubicin against sensitive cell lines. However, the most active compound 12 was not cross-resistant with doxorubicin against KB-V-1.

• 약학회지
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• 제41권6호
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• pp.718-723
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• 1997

In the course of developing novel antitumor intercalating agents, we synthesized 3- carbamoyloxymethyl-azaanthraquinones 6-12, incorporating the latent alkylating functionality. These compounds were designed to explore the effect of heteroatom incorporation into anthraquinone chromophore and the effect of the incorporation of the latent alkylating functionality. The derivatives were prepared by hetero Diels-Alder reaction as a key step followed by functionality of allylic methyl to the desired substituents. Growth inhibitory studies of the azaanthraquinones were conducted in vitro against human cancer cell lines (SNU-354: liver and MCF7: breast) and human epidermoid carcinoma cells that are sensitive (KB-3-1) and multidrug-resistant (KB-V-1). The derivatives were 10 to 100-fold less potent than doxorubicin against sensitive cell lines. However, they were marginally cross-resistant with doxorubicin against KB-V-1.

• 생명과학회지
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• 제30권4호
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• pp.402-409
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• 2020

신경괴사증바이러스(NNV)는 25 nm의 작은 입자 크기에 RNA1 (3.4 kb, RdRp), RNA2 (1.4 kb, capsid protein) 두 가닥의 RNA를 유전정보를 가진다. NNV는 1980년대 말 처음 보고된 이후 전 세계적으로 120여종의 어류에 감염을 일으키며 심각한 피해를 일으키고 있는 바이러스이다. NNV 감염에 의한 피해를 최소화하고 효율적인 백신들을 개발하기 위해서는 무엇보다 NNV 감염에 따른 세포내 신호전달체계를 이해할 필요가 있다. NNV는 세포내 감염 이후 숙주가 가진 바이러스 복제에 필요한 요소들을 이용할 수 있도록 숙주세포의 cell cycle arrest 등의 기작을 이용하는 것으로 알려졌다. 반면에 숙주 세포는 NNV와 감염된 세포를 제어하기 위해 RIG-1-like receptor signaling pathway 등을 통해 NNV 감염을 인지한 다음 IFN signaling pathway를 통해 항바이러스 작용에 필요한 ISG들을 발현시킨다. 또한 감염된 세포들을 사멸시키기 위해 ER stress를 통한 unfolded protein response (UPR), mitochondria-mediated cell death 작용을 통해 감염된 세포의 apoptosis를 유발한다. NNV 감염 기작에 대한 세포신호전달연구는 아직 초기단계이며 검증해야 할 pathway들이 아직도 많이 남아있는 상황이다. 따라서 NNV 감염과 연관된 다양한 세포신호전달체계를 탐색하고 질병 특이적인 세포신호전달체계를 이해함으로써 신속하고 정확한 진단법 및 백신 개발에 많은 도움이 될 것으로 생각된다.

• 치과방사선
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• 제29권1호
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• pp.105-117
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• 1999

Purpose: Ionizing radiations have been reported as an apoptosis initiating stimulus in various cells and it has established that sustained elevations in [Ca/sup 2+/] can lead to DNA fragmentation by Ca/sup 2+/-dependent endonucleases, ultimately resulting in apoptotic cell death. The previous experiments have been reported by using primarily thymocytes and lymphocytes and the change of [Ca/sup 2+/] was measured only by minutes or hours respectively. We need to evaluate [Ca/sup 2+/] in both several minutes and hours after irradiation of radiation of radiation therapy and verify the apoptotic cells. Materials and Methods: We have measured [Ca/sup 2+/] in human gingival epitheloid cancer cell with 10Gy irradiation, at minutely intervals and hourly intervals using digitized video-intensified fluorescence microscopy and the fluorescent Ca/sup 2+/ indicator dye, fura-2. In order to find out that the transient rise in [Ca/sup 2+/] could induced apoptosis, cells were incubated for 1 hour at 37℃ with TdT enzyme, rinsed and resuspended containing fluorescence and observed under a confocal fluorescence microscope. MTT assay was done to determine cell activity and LDH assay was done to determine the amount of necrotic cells. Results: After irradiation, the transient and temporal increasing of [Ca/sup 2+/] in the KB cells was founded. Though, there was no change in the intracellular [Ca/sup 2+/] at 30 minutes and 2 hours after irradiation. We could detect of DNA fragmented cells at 4 hours after 10Gy irradiated cells. There were no significant differences between 4 hour, 1 day, 3 day cells. There were no significant differences in MTT and LDH assay between the irradiated group and the control group after 4 hours and 1 day. Though after 3 days there were differences in MTT and LDH assay between the irradiated group was significantly decreased than the control group, in LDH assay the number of necrotic cell death of the irradiated was higher than the control group. Conclusion: In KB cells there were incipient and temporal increasing of the [Ca/sup 2+/] with 10Gy irradiation and the apoptosis was founded from 4 hours later which was earlier than seeing of the change of the amount of the cellular ability and necrosis.