• Radiation Oncology Journal
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• 제16권4호
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• pp.377-388
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• 1998

목적 : 방사선조사에 의한 apoptosis에서 나타나는 각종 세포주기관련 유전자들의 발현 양상을 RNA와 단백 수준에서 분석하여 방사선조사에 의한 apoptosis에서의 세포주기 조절의 변화를 규명함으로서 방사선치료의 기전에 대한 분자생물학적 이해를 도모하고자 본 연구를 시행하였다. 대상 및 방법 : promyelocytic leukemia 세포주인 HL60 세포주를 배양하여 선형가속기(6MV X-선)로 세포에 8Gy의 방사선을 조사하였다. 조사후 다양한 시간 간격으로 Apoptotic DNA Fragmentation Assay법을 이용하여 apoptosis를 확인하고 동시에 세포주기관련 유전자들(cyclinA, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA와 Rb)을 단백질과 RNA 수준에서 분석하기위해 western blot analysis와 반정량적 RT-PCR을 시행하였다. 결과: 8 Gy의 방사선 조사에 의해 HL60세포에서 apoptosis가 관찰 되었다. 방사선 조사군에서 cyclin A단백은 조사후 48시간까지 시간이 갈수록 증가하였으며, cyclin E, E2F, CDK2 및 Rb 단백은 증가되었다가 다시 감소를 보였다. Rb단백의 증가는 대부분 비활성형인 ppRb (phosphorylated Rb protein)의 양적변화에 의한 것이었다. cyclin Dl, PCNA, COC2, CDK4, $p16^{INK4a}$단백은 발현의 차이를 보이지 않았으며 $p21^{WAF1}$과 $p27^{KIP1}$ 단백은 검출되지 않았다. cyclin A, B, C mRNA는 방사선 조사 직후 감소하였다가 12시간부터 발현이 증가되었으며 cyclin Dl mRNA는 조사후 바로 증가하여 48시간에 다시 감소하였다. cyclin E mRNA는 조사 후 시간이 경과함에 따라 감소하였다. CDK2 mRNA는 3시간째는 감소하다가 6시간부터 많은 증가를 보였으며 CDK4 mRNA는 조사후 6-12시간에 급격한 발현증가를 보였다. $p16^{INK4a}$ RNA는 발현의 변화가 없었으며, $p21^{WAF1}$ 및 $p27^{KIP1}$ RNA의 발현은 관찰되지 않았다. 결론 : 이상의 결과로 미루어볼 때, 방사선 조사에 의한 HL60세포의 apoptosis와 세포의 Gl/S transition는 밀접한 관계가 있는 것으로 생각되며 Rb단백의 증가와 활성형 Rb단백의 감소 현상도 관련이 있는 것으로 사료된다. 이는 E2F의 비정상적인 과발현 및 cyclin E/CDK2의 발현 증가와 관련이 있는 것으로 추측된다. 또한 $p21^{WAF1}$ 및 $p27^{KIP1}$는 방사선에 의한 apoptosis에는 관여되지 않는 것으로 사료된다.

• 한국가금학회지
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• 제27권2호
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• pp.91-98
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• 2000

Phylogenetic tree constructed from the nucleotide sequences of the S1 gene showed that the 15 Korean strains of infectious bronchitis virus(IBV) examined were classified into 2 genetically distinct groups, except one respiratory strain, RB86, which was clustered with Massachusetts group. All the 5 respiratory strains belonged to Korean group I and the rest 9 nephropathogenic strains belonged to Korean group II according to the analysis, based on S1 gene sequences. Like previous classifications corresponded with the geographic origin, Korean stains were discriminated from geographically distinct reference strains of IBV. The nephropathogenic strains within Korean group IIsharing 96% homology were continuously isolated since 1990, and seemed to be genetically stable. Whereas the respiratory strains within Korean group Ⅰ sharing 88% homology were sporadically isolate since 1986m and seemed to be genetically unstable. Because we found putative accumulated point mutation as well as recombination events in Korean group Ⅰ, we discussed why genetic variations have often occurred in respiratory strains rather than nephropathognic strains.

• Journal of Microbiology
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• 제34권4호
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• pp.355-362
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• 1996

Complementation cloning of the argC, E, B, D, F, and G genes in Corynebacterium glutamicum was done by transforming the genomic DNA library into the corresponding arginine auxotrophs fo Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8kb fragments complementing the E. coli argB mutant were also able to complement the E. coli argC, E, A, D, and F mutants, indicating the clustered organization of the arginine biosynthetic genes within the cloned DNA fragments. The insert DNA fragments in the recombinant plasmids, named pRB1 AND pRB2, were physically mapped with several restriction enzymes. By further subcloning the entire DNA fragment containing the functions and by complementation analysis, we located the arg genes in the order of ACEBDF on the restriction map. We also determined the DNA nucleotide sequence of the fragment and report here the sequence of the argB gene. When compared to that with the mutant strain, higher enzyme activity of N-acetylglutamate kinase was detected in the extract of the mutant carrying the plasmid containing the putative argB gene, indicating that the plasmid contains a functional argB gene. Deduced amino acid sequence of the argB gene shows 45%, 38%, and 25% identity to that from Bacillus strearothermophilus, Bacillus substilus, and E. coli respectively. Our long term goal is genetically engineering C. glutamicum which produces more arginine than a wild type strain does.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2012년도 정기총회 및 춘계학술발표회
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• pp.8-8
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• 2012

Ginseng have been traditionally used for strengthening immunity, providing nutrition and recovering health from fatigue. Recently, pharmaceutical activities of ginseng roots have been proven by many researches, and ginseng has become a world-famous medicinal plant. Ginseng saponin, ginsenoside, is one of the most important secondary metabolite in ginseng which has various pharmacological activities. Many studies have aimed to convert major ginsenosides to the more active minor ginsenoside Rg3 for consumer demand ginseng product. Microbial strain GS514 strain was isolated from soil around ginseng roots for enzymatic preparation of ginsenoside Rg3, which strain shows strong ability of converting ginsenoside Rb1and Rd into Rg3 in the solution with NaCl. The gene encoding a ${\beta}$-glucosidase from this GS514 was cloned and expressed in the BL21 (DE3) strain of Escherichia coli. The recombinant enzyme was purified and characterized. The molecular mass of purified was 87.5 kDa, as determined by SDS-PAGE. The gene sequence revealed significant homology to the family 3 glycoside hydrolases. The purified single enzyme also catalyzed the conversion of ginsenoside Rb1 into Rg3. This target enzyme will be able to produce as much saponin for consumer demand ginseng product. Anti-apoptotic proteins bind with pro-apoptotic proteins to induce apoptosis mechanism. Over expression of these anti-apoptotic proteins lead to several cancers by preventing apoptosis. Docking simulations were performed for anti-apoptotic proteins with several ginsenosides from Panax ginseng. Our finding shows ginsenosides particularly Rg3, Rh2 and Rf have more binding affinity with apoptotic proteins. Further, these docking system of each ginsenosides can be extended to experimental screen system for further brief confirmations of several diseases.

• Journal of Ginseng Research
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• 제39권2호
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• pp.105-115
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• 2015

Background: Alcoholic steatosis is the earliest and most common liver disease, and may precede the onset of more severe forms of liver injury. Methods: The effect of Korean Red Ginseng extract (RGE) was tested in two murine models of ethanol (EtOH)-feeding and EtOH-treated hepatocytes. Results: Blood biochemistry analysis demonstrated that RGE treatment improved liver function. Histopathology and measurement of hepatic triglyceride content verified the ability of RGE to inhibit fat accumulation. Consistent with this, RGE administration downregulated hepatic lipogenic gene induction and restored hepatic lipolytic gene repression by EtOH. The role of oxidative stress in the pathogenesis of alcoholic liver diseases is well established. Treatment with RGE attenuated EtOH-induced cytochrome P450 2E1, 4-hydroxynonenal, and nitrotyrosine levels. Alcohol consumption also decreased phosphorylation of adenosine monophosphate-activated protein kinase, which was restored by RGE. Moreover, RGE markedly inhibited fat accumulation in EtOH-treated hepatocytes, which correlated with a decrease in sterol regulatory element-binding protein-1 and a commensurate increase in sirtuin 1 and peroxisome proliferator-activated receptor-a expression. Interestingly, the ginsenosides Rb2 and Rd, but not Rb1, significantly inhibited fat accumulation in hepatocytes. Conclusion: These results demonstrate that RGE and its ginsenoside components inhibit alcoholic steatosis and liver injury by adenosine monophosphate-activated protein kinase/sirtuin 1 activation both in vivo and in vitro, suggesting that RGE may have a potential to treat alcoholic liver disease.

• International Journal of Oral Biology
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• 제32권1호
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• pp.35-43
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• 2007

Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated ${\beta},-galactosidase$, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and $p16^{INK4A}$ protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although $p27^{Kip1}$ and $p15^{INK4B}$, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin ${\alpha}2$, ${\alpha}v$, and ${\beta}1$. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that $p16^{INK4A}/RB$ might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.

• 대한의생명과학회지
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• 제11권4호
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• pp.421-427
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• 2005

To evaluate the diagnostic significance of p16 overexpression in human hepatocellular carcinoma (HCC), we analyzed p16 status and growth inhibitory patterns by p16 overexpression in HCC cell lines having different pRE status. SKHep1 and SNU449 cells show homozygous deletion of p16. The p16 gene in SNU398 cell is inactivated at posttranscription level. Adenovira1-p16 (Ad-p16) infection inhibits the cell growth in Hep3B, SNU398, and SNU449. Failure of growth inhibition in SKHepl results from the low transduction efficiency of adenovirus. The p16-mediated growth inhibition shows G 1 phase arrest in pRE-positive SNU449 but not in pRE-negative Hep3B. These results suggest that therapeutic efficacy of p16 gene might be considered on the transduction efficiency and the toxicity of adenoviral vector. Beside, growth inhibitory effect of p16 could be exerted through either pRE-dependent or -independent pathway.

• 한국물환경학회지
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• 제23권1호
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• pp.27-32
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• 2007

This study focused on estimating the feasibility of a strong basic anion exchanger (PA312OH) as a sorbent for the removal of residual reactive dye and saving chemicals and water. Cellulose reactive dye C.I.RB49 was tested because reactive dye is the largest single group of dyes and that dye needs larger amount of inorganic salts as dyeing agent but nearly 50% of reactive dyes may be lost to the effluent. The adsorption characteristics of PA312OH for C.I.RB49 were as follows. Ion-selectivity among the dye and inorganic salts was Dye > ${SO_4}^{2-}$ > ${CO_3}^{2-}$ > $Cl^-$. C.I.RB49 was exchanged more than 3 times ${SO_4}^{2-}$ and ${CO_3}^{2-}$ and $Cl^-$ was not exchanged absolutely. The exchanging velocity was increased exponentially with increasing temperature. This result is positive effect on treating the high temperature dyeing process wastewater. The exchanged dye percents to initial were 96.8% and 99% at flow rate 20.5 mL/min. and 3.7 mL/min.. The exchanging capacity of PA312OH for C.I.RB49 was 215.2 mg/g at conc.=369.2 mg/L, Temp.=$25^{\circ}C$. 74% inorganic salts were recrystallized from real dark reactive color dyeing wastewater treated with PA312OH.

• 생명과학회지
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• 제11권4호
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• pp.362-370
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• 2001

Regulation of cell proliferation is a complex process involving the regulated expression and /or modification of discrete gene products. which control transition between different stages of the cycle. The purpose of this short review is to provide an overview of somatic cell cycle events and their controls. Cycline have appeared as major positive regulators in this network, because their association to the cyclin-dependent kinases(Cdks) allows the subsequent activation on the Cdk/cyclin complexes and their catalatic activity. In mammalian cells, early to mid G1 progression and late G1 progression leading to S phase entry are directed by D-type cyclins-Cdk4, 6 and cyclin E-Cdk 2 both of which can phosphorylate the retinoblastoma protein (pRB). pRB is a transcriptional repressor which, in its unphosphorylated state, binds to members of the E2F transcription factor family and blocks E2F-dependent transcription of genes controlling the G1 to S phase transition an subsequent DNA synthesis. Cyclin A is produced in late G1 and expressed during S and G2 phae, and expression of B-type cyclins is typically maximal during the G2 to M phase transition and it controls the passage through M phase. They primarily associate with the activate Cdk2, and Cdc2, respectively. On the other hand, the Cdk inhibitors negatively control the activity of C아/cyclin complex by coordinating internal and/or external signals and impending proliferation at several key checkpoints. These current and further findings will provide novel approaches to understanding and treating major diseases.

• 대한화장품학회지
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• 제37권1호
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• pp.75-81
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• 2011

본 연구에서는 유산균 발효에 의한 인삼열매 추출물의 여러 가지 생리활성을 평가하였다. 고성능액체크로마토그래피를 수행하여 인삼열매 추출물과 유산균 발효 인삼열매 추출물의 ginsenoside Re, Rc 및 Rb1의 함량분석 결과 유산균 발효에 의해 ginsenoside Re, Rc 및 Rb1의 함량이 모두 증가하였다. 인삼열매 추출물과 유산균 발효 인삼열매 추출물의 DPPH free radical 소거활성과 SOD 유사활성 측정결과 1.00 %의 농도에서 유산균 발효 인삼열매 추출물은 각각 86.34, 76.82 %의 DPPH free radical 소거활성과 SOD 유사활성을 나타내었고, 인삼열매 추출물은 각각 49.78, 40.80 %의 DPPH free radical 소거활성과 SOD 유사활성을 나타내어 유산균 발효에 의해 항산화 활성이 증가하는 것을 확인하였다. 또한 유산균 발효 인삼열매 추출물 0.50 %의 농도로 처리한 피부 섬유아세포에서 procollagen type I (COL1A1)은 828.13 % 증가하였으며, matrix metalloproteinase (MMP)-1은 87.88 %, tumor necrosis factor (TNF)-${\alpha}$는 99.92 %감소하는 효과를 보여주었다. 이처럼 항산화, 주름, 항염에 효능을 보이는 유산균 발효에 의한 인삼열매 추출물은 기능성 화장품 소재로 개발될 수 있는 가능성이 있다.