• 생명과학회지
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• 제22권4호
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• pp.447-455
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• 2012

Akt는 다양한 세포에서 성장, 발달, 증식, 분화와 같은 생리적 활성에 중요한 역할을 하고 골격근 세포에서 Akt는 재생 및 비대와 위축을 조절한다고 알려져 있다. 골격근 세포의 분화에 있어서 Akt의 역할을 밝히고자 본 연구를 수행하였다. 골격근 세포를 분화 시키기 위해 고밀도 및 저농도의 serum 상태에서 배양하며, 분화된 C2C12 근아세포는 둥근 모양에서 다핵을 가진 긴 모양으로 바뀐다. 이러한 형태학적 변화는 분화 시킨 후 2일부터 일어났다. 또한, 골격근 분화 표지인자인 myogenin D와 myogenin G의 발현은 2일 후 관찰되었다. C2C12 세포주에 Akt1 또는 Akt2의 발현을 저하시키면 이와 더불어 골격근으로의 분화도 저해됨을 확인하였고, 이와는 반대로 Akt1 또는 Akt2를 과발현 시키면 골격근으로 분화가 촉진됨을 알 수 있었다. 이와 더불어 Akt의 활성은 분화유도 2일 후부터 관찰되었고 7일 이후로는 감소하였다. Kruppel-like factor 4의 발현은 6일부터 증가하는 것이 관찰이 되었다. Kruppel-like factor 4의 발현 또한 Akt1 또는 Akt2의 발현양이 감소된 C2C12 근아세포에서 줄어들어 있는 것을 확인하였다. 또한 Kruppel-like factor 4의 프로모터 부위에 대한 전사조절능력이 Akt1 또는 Akt2의 발현을 저하시켰을 때 같이 떨어짐을 확인하였다. 이러한 결과들로 보아 Akt가 골격근 분화를 조절하는데 있어 중요하며, Kruppel-like factor 4 발현이 이를 조절하는 데 있어 중요한 역할을 할 것이라 판단된다.

• Genomics & Informatics
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• 제10권3호
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• pp.167-174
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• 2012

AKT is a signal transduction protein that plays a central role in the tumorigenesis. There are 3 mammalian isoforms of this serine/threonine protein kinase-AKT1, AKT2, and AKT3-showing a broad tissue distribution. We first discovered 2 novel polymorphisms (AKT2 -9826 C/G and AKT3 -811 A/G), and we confirmed 6 known polymorphisms (AKT2 -9473 C/T, AKT2 -9151 C/T, AKT2 -9025 C/T, AKT2 -8618G/A, AKT3 -675 A/-, and AKT3 -244 C/T) of the AKT2 and AKT3 promoter region in 24 blood samples of Korean lung cancer patients using direct sequencing. To evaluate the role of AKT2 and AKT3 polymorphisms in the risk of Korean lung cancer, genotypes of the AKT2 and AKT3 polymorphisms (AKT2 -9826 C/G, AKT2 -9473 C/T, AKT2 -9151 C/T, AKT2 -9025 C/T, AKT2 -8618G/A, and AKT3 -675 A/-) were determined in 360 lung cancer patients and 360 normal controls. Statistical analyses revealed that the genotypes and haplotypes in the AKT2 and AKT3 promoter regions were not significantly associated with the risk of lung cancer in the Korean population. These results suggest that polymorphisms of the AKT2 and AKT3 promoter regions do not contribute to the genetic susceptibility to lung cancer in the Korean population.

• 생명과학회지
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• 제18권12호
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• pp.1651-1656
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• 2008

Redox factor-1 (Ref-1)은 산화적으로 손상된 DNA의 복구와 세포내 산화환원에 민감한 전사인자들의 활성화에 필수적인 역할을 수행한다. 본 연구에서는 마우스 유래 혈관내피세포주 (MAE)에서 nitric oxide (NO) 생성과정에 관여하는 AKT 활성화의 측면에서 adenoviral vector를 사용하여 과발현된 Ref-1의 역할을 살펴보았다. NO 측정을 위하여 fluorophore DAF-2를 사용하였다. 과발현된 Ref-1은 bradykinin으로 자극한 세포뿐만 아니라 자극되지 않은 세포의 NO 생성도 증가시켰다. 놀랍게도 이 과발현된 Ref-1은 AKT의 직접적인 인산화를 유도하였으며, AKT 저해제로 널리 사용되는 wortmannin에 의해 반응이 억제되었다. 또한, Ref-1에 의한 직접적인 AKT 활성화를 증명하기 위하여 HA-tagged activation-deficient AKT를 과발현시키는 adenoviral vector를 사용하였다. 이 방법을 이용한 AKT 활성의 저해는 과발현된 Ref-1에 의한 NO 생성 및 bradykinin 자극에 의한 NO 생성을 억제하였다. 이들 결과는 Ref-1이 마우스 혈관내피세포에서 직접적인 AKT 인산화를 통하여 eNOS 활성화를 유도한다는 것을 의미한다.

• 대한의생명과학회지
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• 제19권3호
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• pp.266-269
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• 2013

Hox genes encode transcription factors important for anterior-posterior body patterning at early stages of embryonic development. However, the precise mechanisms by which signal pathways are stimulated to regulate Hox gene expression are not clear. In the previous study, protein kinase B alpha (Akt1) has been identified as a putative upstream regulator of Hox genes, and Akt1 has shown to regulate Gcn5, a prototypical histone acetyltransferase (HAT), in a negative way in mouse embryonic fibroblast (MEF) cells. Since the activity of HAT such as the CBP/p300, and PCAF (a Gcn5 homolog), was down-regulated by Akt through a phosphorylation at the Akt consensus substrate motif (RXRXXS/T), the amino acid sequence of Gcn5 protein was analyzed. Mouse Gcn5 contains an Akt consensus substrate motif as RQRSQS sequence while human Gcn5 does not have it. In order to see whether Akt1 directly binds to Gcn5, immunoprecipitation with anti-Akt1 antibody was carried out in wild-type (WT) mouse embryonic fibroblast (MEF) cells, and then western blot analysis was performed with anti-Akt1 and anti-Gcn5 antibodies. Gcn5 protein was detected in the Akt1 immunoprecipitated samples of MEFs. This result demonstrates that Akt1 directly binds to Gcn5, which might have contributed the down regulation of the 5' Hoxc gene expressions in wild type MEF cells.

• BMB Reports
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• 제45권9호
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• pp.521-525
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• 2012

In addition to its pivotal role in neuronal survival, PI3K/Akt signaling is integral to neuronal differentiation and neurite outgrowth. However, the exact role of Akt in neuronal differentiation is still controversial. Here, we found that nuclear expression of CA-Akt resulted in unusual rapid neurite outgrowth and overexpression of KD-Akt caused multiple dendrite growth without specific axon elongation. Moreover, microarray data revealed that the expression of FOXQ1 expression was about 10-fold higher in cells with nuclear, active Akt than in control cells. Quantitative real-time PCR analysis showed that mRNA levels were upregulated in NLS-CA-Akt cells as compared to KD or EV cells. Furthermore, our FACS analysis demonstrated that overexpression of NLS-CA-Akt accumulate cells in the G1 phase within 24 h, fitting with the rapid sprouting of neuritis. Thus, our data implied that at least in this early time frame, the overexpression of nuclear, active Akt forced cells into neurite development through probably FOXQ1regulation.

• Asian Pacific Journal of Cancer Prevention
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• 제14권4호
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• pp.2571-2575
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• 2013

AKT1 is a member of the serine/threoine AGC protein kinase family involved in thyroid cancer metabolism, growth, proliferation and survival. It is overexpressed in thyroid tumors. In this study, we designed two AKT1 specific DNAzymes (DRz1 and DRz2) that target AKT1 mRNA. The results showed that DRz1 could decrease the expression of AKT1 by 58%. Furthermore, DRz1 significantly inhibited cell proliferation, induced apoptosis and inhibited invasion in SW597 cells. In addition, down-regulation of survivin expression was associated with decreased caspase-3, VEGF and MMP2 in SW597 cells after 24 h. In our study, the efficacy of DRz1 in decreasing AKT1 protein levels were better than DRz2. AKT1-DRz1 might have anti-tumorigenic activity and may provide the basis for a novel therapeutic intervention in thyroid cancer treatment.

• 약학회지
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• 제48권2호
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• pp.105-110
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• 2004

Akt (or Protein Kinase B; PKB) is a serine/threonine kinase and is activated by phosphoinositide 3-kinase (PI3K) pathway. Recent evidence indicates that the abnormal activities or expression of Akt is closely associated with cancer, diabetes and neuro-degenerative diseases. These findings mean that Akt is likely to be a new therapeutic target for the treatment of disease. Here, we screened the effects of dithiolo-dithione derivatives such as SWU-20004, SWU-20009 and SWU-20025 on Akt activities. Among these compounds, only SWU-20009 (2-Thioxo-[1,3]dithiolo[4,5- $\beta$][1,4]dithiine-5,6-dicarboxylic acid dimethyl ester) inhibited the growth of KATOIII cell at micromolar range of concentration. Further investigation also revealed that SWU-20009 inhibited cellular Akt activity and induced apoptotic cell death.

• 한국응용과학기술학회지
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• 제25권3호
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• pp.370-379
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• 2008

Akt, a serine/threonine protein kinase as a viral oncogene, is a critical regulator of PI3K-mediated cell proliferation and survival. On translocation, Akt is phosphorylated and activated, ultimately resulting in stimulation of cell growth and survival. As a part of our program toward the novel Akt1 inhibitors with potent activity over PI3K signaling pathway, we found primary hit compound 2 with an $IC_{50}$ value of $620\mu}M$ from protein kinase focused library. Based on the structural features of 2, new 1,3,4-thiadiazole derivatives were designed by the introduction of aromatic and heteroaromatic moieties onto thiadiazole nucleus. In this work, a series of 1,3,4-thiadiazole derivatives 1a-1 were synthesized and evaluated for Akt1 inhibitory activity.

• The Korean Journal of Physiology and Pharmacology
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• 제21권5호
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• pp.547-554
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• 2017

Previous studies have demonstrated the role of hydroquinone (HQ), a hydroxylated benzene metabolite, in modulating various immune responses; however, its role in macrophage-mediated inflammatory responses is not fully understood. In this study, the role of HQ in inflammatory responses and the underlying molecular mechanism were explored in macrophages. HQ down-regulated the expression of interferon $(IFN)-{\beta}$ mRNA in LPS-stimulated RAW264.7 cells without any cytotoxicity and suppressed interferon regulatory factor (IRF)-3-mediated luciferase activity induced by TIR-domain-containing adapter-inducing interferon-${\beta}$ (TRIF) and TANK-binding kinase 1 (TBK1). A mechanism study revealed that HQ inhibited IRF-3 phosphorylation induced by lipopolysaccharide (LPS), TRIF, and AKT by suppressing phosphorylation of AKT, an upstream kinase of the IRF-3 signaling pathway. IRF-3 phosphorylation is highly induced by wild-type AKT and poorly induced by an AKT mutant, AKT C310A, which is mutated at an inhibitory target site of HQ. We also showed that HQ inhibited IRF-3 phosphorylation by targeting all three AKT isoforms (AKT1, AKT2, and AKT3) in RAW264.7 cells and suppressed IRF-3-mediated luciferase activities induced by AKT in HEK293 cells. Taken together, these results strongly suggest that HQ inhibits the production of a type I IFN, $IFN-{\beta}$, by targeting AKTs in the IRF-3 signaling pathway during macrophage-mediated inflammation.

• 생명과학회지
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• 제22권1호
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• pp.55-61
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• 2012

Akt는 세포의 증식과 분화에 관여하며 많은 암종에서 과발현되어 있다는 것이 보고되었다. 본 연구에서는 Akt의 조절을 통한 셀레늄의 HT-29 세포의 세포증식억제 시너지효과를 확인하였다. 셀레늄을 농도별과 시간별로 처리하였을 때 HT-29 세포의 증식이 억제되었고, apoptosis가 일어남을 확인하였다. 셀레늄을 농도별로 처리하여 Western blotting 및 immunofluorescence를 실시한 결과 Akt의 인산화가 저해되었고 COX-2의 발현도 저해되었다. 또한 Akt 저해제인 LY294002를 처리한 결과, HT-29 대장암세포의 증식이 억제되었으며, LY294002를 셀레늄과 병행처리하였을 때 셀레늄에 의한 세포증식억제 효과가 더 강하게 나타나는 것을 확인하였다. Akt siRNA에 의한 Akt의 불활성화는 non-transfected 세포에 비하여 HT-29 세포의 성장을 더 강하게 억제하였으며, Akt가 불활성화 되었을 때 COX-2의 발현 역시 non-transfected 세포에 비하여 감소된 것을 확인하였다. 따라서 HT-29 세포에서 셀레늄의 세포증식억제 효과는 Akt와 COX-2 신호분자의 조절을 통해 일어나며, Akt 의 저해는 셀레늄의 대장암세포증식 억제에 시너지 효과를 나타냄을 확인하였다.