• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.188-193
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• 1995

A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSCl13 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or man nose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-$\alpha$-D-glucopyranoside (MeGlc) affected the growth of ZSCl13 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSCl13 cells without pBFT93. It was also found that both $2-deoxy-D-[U-^{14}C]glucose{\;}and{\;}methyl-{\alpha}-D-[U-^{14}C]glucopyranoside$ could be effectively transported into ZSCl13 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.

• Asian Pacific Journal of Cancer Prevention
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• 제16권13호
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• pp.5471-5476
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• 2015

2-deoxy-D-Glucose (2DG) causes cytotoxicity in cancer cells by disrupting thiol metabolism. It is an effective component in therapeutic strategies. It targets the metabolism of cancer cells with glycolysis inhibitory activity. On the other hand, MLN4924, a newly discovered investigational small molecule inhibitor of NAE (NEDD8 activating enzyme), inactivates SCF E3 ligase and causes accumulation of its substrates which triggers apoptosis. Combination of these components might provide a more efficient approach to treatment. In this research, 2DG and MLN4924 were co-applied to breast cancer cells (MCF-7 and SKBR-3) and cytotoxic and apoptotic activity were evaluated the by Micro culture tetrazolium test (MTT), TUNEL and ELISA methods. Caspase3 and Bcl2 genes expression were evaluated by real time Q-PCR methods. The results showed that MLN4924 and MLN4924/2DG dose-dependently suppressed the proliferation of MCF7 and SKBR-3 cells. Cell survival of breast cancer cells exposed to the combination of 2DG/MLN4924 was decreased significantly compared to controls (p<0.05), while 2DG and MLN4924 alone had less pronounced effects on the cells. The obtained results suggest that 2DG/MLN4924 is much more efficient in breast cancer cell lines with enhanced cytotoxicity via inducing a apoptosis cell signaling gene, caspase-3.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3213-3222
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• 2015

Background: Cancer metastasis depends on cell motility which is driven by cycles of actin polymerization and depolymerization. Reactive oxygen species (ROS) and metabolic oxidative stress have long been associated with cancer. ROS play a vital role in regulating actin dynamics that are sensitive to oxidative modification. The current work aimed at studying the effects of sub-lethal metabolic oxidative stress on actin cytoskeleton, focal adhesion and cell migration. Materials and Methods: T47D human breast cancer cells were treated with 2-deoxy-D-glucose (2DG), L-buthionine sulfoximine (BSO), or doxorubicin (DOX), individually or in combination, and changes in intracellular total glutathione and malondialdehyde (MDA) levels were measured. The expression of three major antioxidant enzymes was studied by immunoblotting, and cells were stained with fluorescent-phalloidin to evaluate changes in F-actin organization. In addition, cell adhesion and degradation ability were measured. Cell migration was studied using wound healing and transwell migration assays. Results: Our results show that treating T47D human breast cancer cells with drug combinations (2DG/BSO, 2DG/DOX, or BSO/DOX) decreased intracellular total glutathione and increased oxidized glutathione, lipid peroxidation, and cytotoxicity. In addition, the drug combinations caused a reduction in cell area and mitotic index, prophase arrest and a decreased ability to form invadopodia. The formation of F-actin aggregates was increased in treated T47D cells. Moreover, combination therapy reduced cell adhesion and the rate of cell migration. Conclusions: Our results suggest that exposure of T47D breast cancer cells to combination therapy reduces cell migration via effects on metabolic oxidative stress.

• 미생물학회지
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• 제30권3호
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• pp.154-159
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• 1992

섬유소 분해능이 우수한 P. verruculosum 의 분생포자로부터 원형질체의 생성 및 재생에 대하여 조사하였다. 균의 원형질체 생성을 위한여 2-deoxy-D-glucose(2-DG) 가 첨가된 최소배지에서 분생포자를 10-12 시간 전배양 시킨후 시판 세포벽분해효소들을 작용시켜 본 결과 그중 Novozyme 234 (1%, w/v) 에서 50% 이상의 가장 좋은 원형질체 생성 수율을 보였다. 전배양 되지 않은 포자로부터는 원형질체 생성이 이루어지지 않았으며 원형질체 생성율을 향상시키기 위한 첨가테로서 2-DG 의 효과가 무첨가에 비해 2배 이상의 수율을 증가시켰다. 포자 원형질체의 재생율은 49.2% 로 동 균주의 균사체 원형질체의 재생율4.6-27.8%(삼투압 조절체로서 0.6 M magnesium sulfate) 보다 높았다. 원형질체 생성 및 재생을 위한 상투압 안정제로서는 0.6M ammonium sulfate 와0.6 M magnesium sulfate가 가장 적절하였다. 광학 형미경을 통하여 부풀린 포자로부터 원형질체의 생성과정과 두 가지 각기 다른 양상의 재생과정이 관찰되었다.

• Toxicological Research
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• 제24권4호
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• pp.245-251
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• 2008

Dietary restriction (DR) is the most efficacious intervention for retarding the deleterious effects of aging. DR increases longevity, decreases the occurrence and severity of age-related diseases, and retards the physiological decline associated with aging. The beneficial effects of DR have been mostly studied in non-neuronal tissues. However, several studies have showed that DR attenuate neuronal loss after several different insults including exposure to kainate, ischemia, and MPTP. Moreover, administration of the non-metabolizable glucose analog 2-deoxy-D-glucose (2DG) could mimic the neuroprotective effect of DR in rodent, presumably by limiting glucose availability at the cellular level. Based on the studies of chemically induced DR, it has been proposed that the mechanism whereby DR and 2DG protect neurons is largely mediated by stress response proteins such as HSP70 and GRP78 which are increased in neurons of rats and mice fed a DR regimen. In addition, DR, as mild metabolic stress, could lead to the increased activity in neuronal circuits and thus induce expression of neurotrophic factors. Interestingly, such increased neuronal activities also enhance neurogenesis in the brains of adult rodents. In this review, we focus on what is known regarding molecular mechanisms of the protective role of DR in neurodegenerative diseases and aging process. Also, we propose that DR is a mild cellular stress that stimulates production of neurotrophic factors, which are major regulators of neuronal survival, as well as neurogenesis in adult brain.

• 대한핵의학회지
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• 제29권3호
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• pp.294-306
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• 1995

국소뇌허혈에서 MK-801의 신경세포방어효과를 검토하기 위하여 흰쥐의 중뇌동맥을 폐색시켜 뇌허혈 및 경색을 유발하기 전과 후에 MK-801을 투여한 후 국소뇌포도당이용률의 변화를 2-deoxy-D-(14C)glu-cose 자가방사기록법을 이용하여 측정하고 이를 대조군과 비교하였다. 중대뇌동맥폐색 시술 전과 후에 평균동맥압, 직장체온, 동맥혈 PH, $pCO_2,\;pO_2$의 유의한 차이는 없었다. [$^{14}C$]DG 실험 중 대조군, MK-801 전처처군과 MK-801 후처치군 사이에서 평균동맥압, 직장 체온, 동맥혈장 당농도, 동맥혈 pH, $pCO_2,\;pO_2$의 유의한 차이는 없었다. 대조군에서 중대뇌동맥폐색 24시간 후에 폐색된 대뇌반구의 전두엽피질, 두정엽피질, 담창구, 편도, 해마 CA-2 지역, CA-3 지역, 내비회, 후두엽피질, 측두엽피질의 국소뇌포도당이용률은 반대쪽정상 대뇌반구 상동부위의 국소뇌포도당이용률보다 유의하게 낮았다. MK-801 전처치군에서 전두엽피질, 두정엽피질, 편도, 해마 CA-2 지역, 내비회, 후두엽피질, 측두엽피질의 병변/정상 국소뇌포도당이용률 비는 대조군의 그것보다 유의하게 높았다. 한편 MK-801 후처치군에서는 전두엽피질, 두정엽피질, 해마 CA-2 지역, 후두엽피질, 측두엽피질의 병변/정상 국소뇌포도 당이용률 비가 대조관의 그것보다 유의하게 높았다. MK-801 전처치군 정상 대뇌반구의 미상핵-피각, 전두엽피질, 두정엽피질, 대상회전, 담창구, 시상, 뇌량, 해마 CA-1, CA-2, CA-3 지역, 후두엽피질, 측두엽피질의 국소뇌포도당이용률은 대조군 정상 대뇌반구의 그것보다 유의하게 높았다. 반면 MK-801 후처치군 정상 대뇌반구의 국소뇌포도당이용률은 대조군 정상대뇌반구의 국소뇌포도당이용률과 유의한 차이가 없었다. 뿐만 아니라 폐색된 대뇌반구에서도 정상 대뇌반구와 매우 유사한 결과를 보였다. 즉, MK-801 전처치군에서는 미상핵-피각, 전두엽피질, 두정엽피질, 대상회전, 담창구, 시상, 뇌량, 해마 CA-1, CA-2, CA-3 지역, 내비회, 후두엽피질, 측두엽피질의 국소뇌포도당이용률이 대조군보다 유의하게 높은 반면, MK-801 후처치군에서는 대조군과 유의한 국소뇌포도당이용률의 차이를 나타내지 않았다. 이상의 결과로부터 국소뇌허혈에서 MK-801은 신경세포방어효과를 나타낼 것으로 생각된다. MK-801은 뇌포도당이용률을 국소적으로 증가시키며, 이는 NMDA 수용체 길항제 투여와 뇌기능 변화의 유관성을 시사하는 것이다.