• 대한의생명과학회지
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• 제10권4호
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• pp.325-332
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• 2004

Inflammatory factor such as Interleukin-1 play important roles in determining the fate of both acute and chronic neurological disorders. We investigated whether inhibitors of PKC or PTK can serve as pharmacological agents to reduce IL-I production and the mechanisms underlying their pharmacological effects in a mixed population of glia. Inhibitors of PKC such as H7, Go6976 and Ro31-8220 significantly reduced both the mRNA and protein levels of IL-1α and IL-β in lipopolysaccharide-activated primary glial cells. While the PTK inhibitor genistein also significantly reduced the production of these cytokines, it did not affect the expression of their mRNA. Taken together, inhibitors of PKC and PTK could serve as pharmacological agents to reduce IL-1 production. However, the mechanisms underlying their pharmacological effects are different. Our results provide evidence that inhibitors of protein kinases can serve as pharmacological agents to modulate IL-1 production in glial cell, and in turn, alleviate neuronal injury.

• Journal of Microbiology and Biotechnology
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• 제22권4호
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• pp.571-573
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• 2012

Glyceraldehyde-3-phosphate (G-3-P), the substrate of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), is a key intermediate in several metabolic pathways. Recently, we reported that G-3-P directly inhibits caspase-3 activity in a reversible noncompetitive mode, suggesting the intracellular G-3-P level as a cell fate decision factor. It has been known that apoptotic stimuli induce the generation of NO, and NO S-nitrosylates GAPDH at the catalytic cysteine residue, which confers GAPDH the ability to bind to Siah-1, an E3 ubiquitin ligase. The GAPDH-Siah-1 complex is translocated into the nucleus and subsequently triggers the apoptotic process. Here, we clearly showed that intracellular G-3-P protects GAPDH from S-nitrosylation at above a certain level, and consequently maintains the cell survival. In case G-3-P drops below a certain level as a result of exposure to specific stimuli, G-3-P cannot inhibit S-nitrosylation of GAPDH anymore, and consequently GAPDH translocates with Siah-1 into the nucleus. Based on these results, we suggest that G-3-P functions as a molecule switch between cell survival and apoptosis by regulating S-nitrosylation of GAPDH.

• 생체재료학회지
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• 제22권4호
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• pp.337-345
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• 2018

Background: Human mesenchymal stem cells (hMSCs) are, due to their pluripotency, useful sources of cells for stem cell therapy and tissue regeneration. The phenotypes of hMSCs are strongly influenced by their microenvironment, in particular the extracellular matrix (ECM), the composition and structure of which are important in regulating stem cell fate. In reciprocal manner, the properties of ECM are remodeled by the hMSCs, but the mechanism involved in ECM remodeling by hMSCs under topographical stimulus is unclear. In this study, we therefore examined the effect of nanotopography on the expression of ECM proteins by hMSCs by analyzing the quantity and structure of the ECM on a nanogrooved surface. Methods: To develop the nanoengineered, hMSC-derived ECM, we fabricated the nanogrooves on a coverglass using a UV-curable polyurethane acrylate (PUA). Then, hMSCs were cultivated on the nanogrooves, and the cells at the full confluency were decellularized. To analyze the effect of nanotopography on the hMSCs, the hMSCs were re-seeded on the nanoengineered, hMSC-derived ECM. Results: hMSCs cultured within the nano-engineered hMSC-derived ECM sheet showed a different pattern of expression of ECM proteins from those cultured on ECM-free, nanogrooved surface. Moreover, hMSCs on the nano-engineered ECM sheet had a shorter vinculin length and were less well-aligned than those on the other surface. In addition, the expression pattern of ECM-related genes by hMSCs on the nanoengineered ECM sheet was altered. Interestingly, the expression of genes for osteogenesis-related ECM proteins was downregulated, while that of genes for chondrogenesis-related ECM proteins was upregulated, on the nanoengineered ECM sheet. Conclusions: The nanoengineered ECM influenced the phenotypic features of hMSCs, and that hMSCs can remodel their ECM microenvironment in the presence of a nanostructured ECM to guide differentiation into a specific lineage.

• Biomolecules & Therapeutics
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• 제32권3호
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• pp.267-280
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• 2024

Apoptosis, programmed cell death pathway, is a vital physiological mechanism that ensures cellular homeostasis and overall cellular well-being. In the context of cancer, where evasion of apoptosis is a hallmark, the overexpression of anti-apoptotic proteins like Bcl2, Bcl-xL and Mcl-1 has been documented. Consequently, these proteins have emerged as promising targets for therapeutic interventions. The BCL-2 protein family is central to apoptosis and plays a significant importance in determining cellular fate serving as a critical determinant in this biological process. This review offers a comprehensive exploration of the BCL-2 protein family, emphasizing its dual nature. Specifically, certain members of this family promote cell survival (known as anti-apoptotic proteins), while others are involved in facilitating cell death (referred to as pro-apoptotic and BH3-only proteins). The potential of directly targeting these proteins is examined, particularly due to their involvement in conferring resistance to traditional cancer therapies. The effectiveness of such targeting strategies is also discussed, considering the tumor's propensity for anti-apoptotic pathways. Furthermore, the review highlights emerging research on combination therapies, where BCL-2 inhibitors are used synergistically with other treatments to enhance therapeutic outcomes. By understanding and manipulating the BCL-2 family and its associated pathways, we open doors to innovative and more effective cancer treatments, offering hope for resistant and aggressive cases.

• 한국토양비료학회지
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• 제25권4호
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• pp.387-393
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• 1992

접종근류균(接種根瘤菌)의 소장(消長)을 유전공학적(遺傳工學的)으로 조제(調劑)한 Bradyrhizobium japonicum RJB6 $str^rnal^rneo^r$을 이용(利用)하여 연구(硏究)하였다. 전남(全南)일대로 부터 분리(分離) 동정한 균(菌)에 먼저 streptomycin과 nalidixic acid에 저항성(抵抗性)을 갖는 spontaneous mutant를 유도하고 이어 pSUP2021를 함유(含有)하고 있는 E. coli와 conjugation하여 neomycin 저항성(抵抗性) gene(Tn5)을 도입(導入)하였다. Southern hybridization한 결과(結果) 4.9kb상에서 Tn5를 확인(確認)했다. 서로 다른 균밀도(菌密度)로 파종(播種)한 40일후, 크로로필을 제외한 근류수(根瘤數), 근류신선중(根瘤新鮮重), 간장 및 질소함량이 low cell suspension 처리구보다 heavy cell suspension 처리구에서 약간 높았다. 표식균주(標識菌株)의 회수율(回收率)은 heavy cell suspension 처리구에서 12%인 반면, low cell suspension 처리구에서는 5%에 지나지 않았다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.56-56
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• 2003

Beta-catenin is very important in early development including involvement in cell adhesion, cell signaling, and developmental fate specification. Cell-cell interaction is an important process during mammalian embryonic development. In preimplantation embryos, embryonic compaction is the process of increased cellular flattening and adhesion of junctional complexes and results in a polarized distribution. Beta-catenin is associated with embryonic compaction in mammals. Here, we examined the relationship between beta-catenin expression and compaction in porcine embryos derived from in vitro fertilization. First of all, we investigated beta-catenin expression in each embryonic developmental stage and also focused on expression pattern according to full, partial and non-compaction at morula stage. We used the immunocyto-chemical method in this research. To confirm compaction affects on the embryonic development, we compared between compaction and developmental rates to the blastocyst. The result showed that compaction and non-compaction rates were 14.6% and 63.8% at 4 days after IVF, respectively The developmental rates to the blastocyst and their total cell number were 50.9% vs 36.4% and 41.4$\pm$11.5 vs 26.8$\pm$12.7 in compaction and non-compaction groups. Although no difference was detected in the ratio of ICM to total cells between two groups, total cell number of the blastocysts in compaction group was superior to that of the blastocysts in non-compaction group (P<0.05). Expression of beta-catenin appeared in the boundary of membrane surface between blastomeres in 2- and 4-cell stage, and observed irregular pattern from 8-cell to blastocyst stage. We also investigated beta-catenin expression pattern according to the degree of compaction in the 3 groups; full, partial (>50%) and non-compaction. The expression signal in fully compacted embryos was stronger than those of partial and non-compacted embryos. Especially, beta-catenin expression appeared various patterns in morula stage suggesting the aberrant distribution of beta-catenin is affected by compaction patterns. Our results suggest that abnormal beta-catenin expression was affected by embryo quality and further development in porcine embryos in vitro.

• Molecules and Cells
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• 제39권4호
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• pp.352-357
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• 2016

Vertebrate neurogenesis requires inhibition of endogenous bone morphogenetic protein (BMP) signals in the ectoderm. Blocking of BMPs in animal cap explants causes the formation of anterior neural tissues as a default fate. To identify genes involved in the anterior neural specification, we analyzed gene expression profiles using a Xenopus Affymetrix Gene Chip after BMP-4 inhibition in animal cap explants. We found that the xCyp26c gene, encoding a retinoic acid (RA) degradation enzyme, was upregulated following inhibition of BMP signaling in early neuroectodermal cells. Whole-mount in situ hybridization analysis showed that xCyp26c expression started in the anterior region during the early neurula stage. Overexpression of xCyp26c weakly induced neural genes in animal cap explants. xCyp26c abolished the expression of all trans-/cis-RA-induced posterior genes, but not basic FGF-induced posterior genes. Depletion of xCyp26c by morpholino-oligonucleotides suppressed the normal formation of the axis and head, indicating that xCyp26c plays a critical role in the specification of anterior neural tissue in whole embryos. In animal cap explants, however, xCyp26c morpholinos did not alter anterior-to-posterior neural tissue formation. Together, these results suggest that xCyp26c plays a specific role in anterior-posterior (A-P) neural patterning of Xenopus embryos.

• Reproductive and Developmental Biology
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• 제37권4호
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• pp.219-224
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• 2013

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

• The Korean Journal of Physiology and Pharmacology
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• 제12권4호
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• pp.185-191
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• 2008

Activation of c-Jun N-terminal kinase (JNK), a member of the mitogen-activated protein kinase family, is an important cellular response that modulates the outcome of the cells which are exposed to the tumor necrosis factor (TNF) or the genotoxic stress including DNA damaging agents. Although it is known that JNK is activated in response to genotoxic stress, neither the pathways to transduce signals to activate JNK nor the primary sensors of the cells that trigger the stress response have been identified. Here, we report that the receptor interacting protein (RIP), a key adaptor protein of TNF signaling, was required to activate JNK in the cells treated with certain DNA damaging agents such as adriamycin (Adr) and 1-${\beta}$-D-arabinofuranosylcytosine (Ara-C) that cause slow and sustained activation, but it was not required when treated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) and short wavelength UV, which causes quick and transient activation. Our findings revealed that this sustained JNK activation was not mediated by the TNF (tumor necrosis factor) receptor signaling, but it required a functional ATM (ataxia telangiectasia) activity. In addition, JNK inhibitor SP-600125 significantly blocked the Adr-induced cell death, but it did not affect the cell death induced by MNNG. These findings suggest that the sustained activation of JNK mediated by RIP plays an important role in the DNA damage-induced cell death, and that the duration of JNK activation relays a different stress response to determine the cell fate.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1063-1067
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• 2007

Bcl-2 family 단백질은 세포사 조절에 매우 중요한 역할을 하며 세포사 촉진 Bcl-2 family 단백질과 세포사 억제 Bcl-2 family 단백질 사이의 균형적인 상호작용이 세포의 운명을 결정하는 주요인자이다. Bcl-2 family 단백질 중 하나인 Noxa 단백질은 p53 에 의한 전사되는 단백질로 처음 발견되었다. Noxa 단백질이 어떻게 세포사를 조절하는지를 이해하기 위해 Yeast two-hybrid 방법을 통해 Noxa 단백질과 결합하는 파트너 단백질을 검색하였고 이를 통해 세포사 억제 단백질 중 하나인 Mcl-1를 발견하였다. 사람 대장암 세포주인 HCT 116에서 Noxa 단백질과 Mcl-1 단백질이 결합하는 것을 면역침전 방법을 통하여 확인하였다. HCT 116 세포주에서 Mcl-1 단백질 과다발현은 Noxa에 의한 세포사 유도를 크게 억제하였다. Noxa 단백질 과다발현에 의한 세포사 과정에서 Mcl-1 단백질이 분해되는 것을 발견하였고 이는 caspase 억제제인 z-VAD-fmk에 의해서 억제되었다. 이는 Mcl-1 단백질이 cas-pase에 의해서 분해되는 것으로 간주된다. 결론적으로, Noxa와 Mcl-1의 결합은 세포사 과정 중 caspase에 의한 Mcl-1 단백질 분해를 유도를 매개할 수 있을 것으로 추측된다.