• 한국발생생물학회지:발생과생식
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• 제15권2호
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• pp.113-120
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• 2011

BCL-2 family 단백질들은 세포사멸 신호전달 체계에서 중추적인 역할을 수행하는 것으로 알려져 있으며, BCL2L10 단백질은 그 중 하나로 세포의 사멸과 생존을 조절하는 것으로 알려져 있다. 특이하게도 BCL2L10 단백질은 세포 또는 조직 특이적으로 서로 상반되는 친 세포사멸 또는 항 세포사멸 효과가 각각 보고되어 있다. 현재까지 난소세포에서의 BCL2L10의 발현 여부 및 기능은 알려져 있지 않다. 따라서 본 연구에서 인간 난소 과립세포주인 KGN 세포에서의 BCL2L10 단백질의 발현 여부를 확인한 결과, 상당한 양의 단백질이 발현함을 확인할 수 있었으며, 또한 세포사멸효과를 확인하기 위해서 BCL2L10 단백질을 dose-dependent하게 과발현시킨 후 세포의 생존에 미치는 영향을 분석한 결과,BCL2L10은 KGN 세포에서 과발현 시 세포사멸을 유도함을 관찰하였다. BCL2L10 단백질을 과발현 시 Caspase 9와 3를 활성화 하였으며, 면역염색법을 통해서 BCL2L10 단백질이 미토콘드리아에 위치하는 것을 확인하였다. 또한 BCL2L10단백질의 과발현에 의해 미토콘드리아에서 cytochrome c가 세포질로 분비되는 것을 확인하였다. 이상의 결과로서 본 연구는 BCL2L10의 과발현이 KGN 세포에서 세포사멸을 유도하고, 또한 미토콘드리아에 위치하여 세포질로 cytochrome c를 분비하여 Caspase 9와 3을 활성화 시키는 메커니즘으로 세포사멸을 유도함을 확인하였다. 이러한 연구결과는 BCL2L10단백질이 인간 난소과립세포의 생존과 사멸을 조절하는 인자임을 최초로 규명한 것으로서, 추후 난소에서의 BCL2L10단백질의 생리적인 기능 및 신호 조절 연구의 기반 데이터로서 그 의의가 있으며 활용될 수 있다.

• 한국발생생물학회지:발생과생식
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• 제15권1호
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• pp.61-69
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• 2011

Previously, we have shown that Bcl2l10 as a member of Bcl-2 family, key regulators of the apoptotic process, is dominantly expressed in oocytes of ovary but several member of the Bcl-2 family are not expressed in oocytes. Recent our studies had been processed about roles and regulatory mechanisms of Bcl2l10 in oocytes. Microinjection of Bcl2l10 RNAi into the cytoplasm of germinal vesicle oocytes resulted in metaphase I (MI) arrest and exhibited abnormalities in their spindles and chromosome configurations (Yoon et al., 2009). The present study was conducted to elucidate the downstream genes regulated by Bcl2l10 and signaling networks in Bcl2l10 RNAi microinjected oocytes by using microarray analysis. Surprisingly, we found that a large proportion of genes regulated by Bcl2l10 RNAi were involved in the cell cycle and actin skeletal system regulation as important upstream genes of Bcl2l10. Among the transcripts with highly significant fold changes more than 2-fold, Tpx2 and Cep192 are 16.1- and 8.2-fold down regulated respectively by Bcl2l10 RNAi. Tpx2 and Cep192 are known as cofactors that control Aurora A kinase activity and localization. Therefore, we concluded that Bcl2l10 may have important roles during oocyte meiosis as functional upstream regulator of Tpx2 and Cep192.

• 생명과학회지
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• 제13권1호
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• pp.118-127
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• 2003

Phenylalanine의 구조유사체인 p-fluotophenylalanine (FPA)은 인체 급성백혈병세포주인 Jurkat T 세포의 세포자살을 유도한다. FPA에 의한 세포자살에 미치는 Bcl-2 또는 Bcl-xL의 영향을 조사하기 위해, Bcl-2 또는 Bcl-xL을 stable transfection하거나 empty vectors만을 Transfection한 Jurkat 세포를 이용하여 FPA의 세포독성과 FPA에 의한 세포내 세포자살 신호전달경로를 비교 분석하였다. Jurkt T 세포에 0.63∼3.0 mLf의 FPA를 처리하였을 때 세포의 생육도는 농도에 비례하여 감소하였다. 또한 세포자살관련 DNA fragmentation, caspase-8 activatoin, Bid cleavage, mitochondria로 부터의 cytochrome c 방출, caspase-9 및 -3 activation, PARP degradation 등이 유도되었다. 한편, FPA에 의해 유도되는 이러한 일련의 생화학적 현상들은 Bcl-2 또는 Bcl-xL의 overexpression에 의해 현저히 저해되었다. 이상의 결과들은 caspase-8 activation, Bid cleavage, mitochondnal cytochrome c 방출에 의해 활성화되는 casuase cascade 등의 현상이, Bcl-2 또는 Bcl-xL에 의해 억제됨을 나타내며 FPA에 의해 유도되는 세포자살에 필요한 과정임을 시사한다.

• The Korean Journal of Physiology and Pharmacology
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• 제14권1호
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• pp.15-20
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• 2010

It has been shown that CA repeats in the 3'-untranslated region (UTR) of bcl-2 mRNA contribute the constitutive decay of bcl-2 mRNA and that hnRNP L (heterogenous nuclear ribonucleoprotein L) interacts with CA repeats in the 3'-UTR of bcl-2 mRNA, both in vitro and in vivo. The aim of this study was to determine whether the alteration of hnRNP L affects the stability of bcl-2 mRNA in vivo. Human breast carcinoma MCF-7 cells were transfected with hnRNP L-specific shRNA or hnRNP L-expressing vector to decrease or increase hnRNP L levels, respectively, followed by an actinomycin D chase. An RT-PCR analysis showed that the rate of degradation of endogenous bcl-2 mRNA was not affected by the decrease or increase in the hnRNP L levels. Furthermore, during apoptosis or autophagy, in which bcl-2 expression has been reported to decrease, no difference in the degradation of bcl-2 mRNA was observed between control and hnRNP L-knock down MCF-7 Cells. On the other hand, the levels of AUF-1 and nucleolin, transacting factors for ARE in the 3'UTR of bcl-2 mRNA, were not significantly affected by the decrease in hnRNP L, suggesting that a disturbance in the quantitative balance between these transacting factors is not likely to interfere with the effect of hnRNP L. Collectively, the findings indicate that the decay of bcl-2 mRNA does not appear to be directly controlled by hnRNP L in vivo.

• Clinical and Experimental Reproductive Medicine
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• 제47권3호
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• pp.155-160
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• 2020

Objective: The genes Bcl-2 and Bax play important roles in apoptosis. Many studies have shown that formalin has a strong deleterious effect on male fertility and can induce apoptosis. L-carnitine has been reported to potentially reverse the negative effects of formalin, leading to improved spermatogenesis. In this study, we examined the levels of expression of Bcl-2 and Bax in mice treated with formalin and L-carnitine. Methods: Thirty adult BALB/c mice were categorized into three groups. The mice in the control group (n = 10) were not injected with any substance. The mice in the second group (n = 10) received 10 mg/kg of formalin daily via an intraperitoneal injection, while those in the final group (n = 10) were intraperitoneally injected daily with a dose of 10 mg/kg of formalin and 100 mg/kg of L-carnitine. All mice were kept in isolated cages for 31 days. Results: The expression of Bax was significantly higher in the formalin-treated mice than in the mice of the control group, while the expression of Bcl-2 was significantly lower in the formalin-treated mice than in the control mice. Additionally, relative to control mice, Bcl-2 expression increased and Bax expression decreased in the mice administered both formalin and L-carnitine. Conclusion: In this study, L-carnitine was shown to augment Bcl-2 expression and to reduce Bax expression, indicating that this compound may inhibit apoptosis. Due to its positive effects, L-carnitine can be used as a prophylactic treatment for people who routinely come into direct contact with formalin as an occupational hazard.

• Biomolecules & Therapeutics
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• 제26권5호
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• pp.446-457
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• 2018

Adiponectin, a hormone predominantly originated from adipose tissue, has exhibited potent anti-inflammatory properties. Accumulating evidence suggests that autophagy induction plays a crucial role in anti-inflammatory responses by adiponectin. However, underlying molecular mechanisms are still largely unknown. Association of Bcl-2 with Beclin-1, an autophagy activating protein, prevents autophagy induction. We have previously shown that adiponectin-induced autophagy activation is mediated through inhibition of interaction between Bcl-2 and Beclin-1. In the present study, we examined the molecular mechanisms by which adiponectin modulates association of Bcl-2 and Beclin-1 in macrophages. Herein, we demonstrated that globular adiponectin (gAcrp) induced increase in the expression of AUF1 and ZFP36L1, which act as mRNA destabilizing proteins, both in RAW 264.7 macrophages and primary peritoneal macrophages. In addition, gene silencing of AUF1 and ZFP36L1 caused restoration of decrease in Bcl-2 expression and Bcl-2 mRNA half-life by gAcrp, indicating crucial roles of AUF1 and ZFP36L1 induction in Bcl-2 mRNA destabilization by gAcrp. Moreover, knock-down of AUF1 and ZFP36L1 enhanced interaction of Bcl-2 with Beclin-1, and subsequently prevented gAcrp-induced autophagy activation, suggesting that AUF1 and ZFP36L1 induction mediates gAcrp-induced autophagy activation via Bcl-2 mRNA destabilization. Furthermore, suppressive effects of gAcrp on LPS-stimulated inflammatory mediators expression were prevented by gene silencing of AUF1 and ZFP36L1 in macrophages. Taken together, these results suggest that AUF1 and ZFP36L1 induction critically contributes to autophagy induction by gAcrp and are promising targets for anti-inflammatory responses by gAcrp.

• BMB Reports
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• 제45권5호
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• pp.287-292
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• 2012

FGF-2 is involved in cell survival, proliferation, apoptosis, and angiogenesis in a wide variety of cells. FRGRs, PI3K and MAP kinases are well known mediators of FGF signaling. Despite its known roles during many developmental processes, including osteogenesis, there are few known targets of FGF-2. In the present study, we identified Bcl2-A1 and Bcl-xL as two prominent targets involved in promoting cell survival. Pretreatment of ATDC5 cells with FGF-2 increased cell survival, while siRNAs specific for Bcl2-A1 and Bcl-xL compromised the anti-apoptotic effect of FGF-2, sensitized the cells to apoptosis triggered by TNF-${\alpha}$. Chemical inhibition of FGFR, NFkB, and PI3K activity by PD173074, pyrrolidine dithiocarbamate, and LY294002 respectively abrogated the FGF-2-mediated induction of Bcl2-A1 and Bcl-xL expression. Taken together, our data demonstrate that a subset of Bcl2 family proteins are the targets of FGF-2 signaling that promotes the survival of ATDC5 cells.

• BMB Reports
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• 제35권5호
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• pp.452-458
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• 2002

Exposure to ultraviolet light (UV) induces apoptosis in mammalian cells, The caspase group of proteases is required for the appotosis. This pathway is initiated by a release of cytochrome c from the mitochondria into the cytosol. Several Bcl-2 family proteins can regulate the release of cytochrome c by stabilizing the mitochondrial membrane. Here we show that expression of the endogenous bcl-xL was strongly downregulated in NIH3T3 cells within 2 h after UV-C irradiation, and that of bax was upregulated from 8 h after irradiation. Apoptosis was induced in more than 50% of the NIH3T3 cells 48 h after irradiation. Constitutive overexpression of bcl-xL in NIH3T3 cells protected the UV-induced apoptosis by preventing the loss of mitochondrial membrane potential and the activation of caspase 9. There results suggest that downregulation of Bcl-xL is relevant to UV-induced apoptosis of tibroblasts.

• Journal of Plant Biotechnology
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• 제38권3호
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• pp.209-214
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• 2011

본 실험에서는 상추 연산홍에 apoptosis 관련 유전자인 Bcl-2 유전자를 도입하여 내병성 상추의 육성을 하기 위한 목적으로 형질전환을 수행하였다. 상추의 자엽조직을 NPTII-35S Promoter와 Bcl-2 유전자가 삽입된 Agrobacterium GV 3101과 공동배양 한 후 0.1 mg/L NAA, 0.5 mg/L BAP, 100 mg/L Kanamycin, 300 mg/L Lilacillin이 첨가된 MS 배지에서 식물체가 유기되었다. Kanamycin 내성을 가진 식물체들을 PCR, Southern blot 분석을 통해 Bcl-2 유전자가 안정적으로 식물체 genome 안에 삽입되었음을 확인하였다. 100개의 형질전환식물체가 확인되었으며 T1식물체를 채종하였다. T1 종자를 파종하여 Sclerotinia sclerotiorum. 균주를 접종하여 내병성 검정을 실시하였고 그 중 2개의 line에서 내병성을 확인하였다. 이러한 결과를 통하여 인간의 apoptosis에 관련하는 유전자가 식물체 내에서 안정된 발현을 통하여 내병성을 증가시켰음을 밝혔다.

• 동아시아식생활학회지
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• 제24권6호
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• pp.739-747
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• 2014

본 연구는 인체유방암세포인 MDA-MB-231세포를 이용하여 IDE를 처리하였을 때 세포사멸에 미치는 영향을 세포화학적 방법으로 확인하였다. IDE를 각각 0, 20, 30 및 $40{\mu}g/mL$ 첨가하여 24시간 배양한 후, 세포증식 억제, 막투과성, 세포내 ROS 분석 및 세포사멸 단계의 특성을 FACS 분석하고, RT-PCR에 의한 사멸관련 유전자 중 Bax/Bcl-2 ratio 분석을 통하여 IDE의 항암작용의 조절작용을 밝히고자 하였다. MTT의 세포 증식 억제는 첨가된 IDE의 농도에 따라 유의적으로 감소하였다(p<0.05). 이와 동시에, trypan blue에 대한 염색성과 DCF-DA 형광 분석의 결과는 각각, 막투과성과 세포내 ROS 농도가 모두 농도 의존적으로 증가됨을 보였다(p<0.05). 이와 같은 IDE 농도에 따른 세포화학적 변화 중에서 세포의 초기사멸에서 후기사멸 과정으로 급격한 사멸단계의 변화가 특히, IDE 농도 30과 $40{\mu}g/mL$에서 일어났다. RT-PCR 분석에 의한 Bax/Bcl-2 ratio도 IDE 농도 30과 $40{\mu}g/mL$에서 급격히 증가하였다(p<0.05). 이와 같은 세포화학적 결과와 RT-PCR 결과를 종합해 볼 때, IDE의 유방암세포(MDA-MB-231)에 대한 세포사멸작용은 막 투과성의 증가와 ROS 증가를 통하여 세포에 점진적인 손상을 일으키며, 이는 세포의 생화학적 변화도 초래하여 세포 증식 억제를 감소시켜, 결국 세포내의 사멸관련 유전자 지표인 Bax/Bcl-2 ratio를 크게 변화시키는 일련의 세포사멸을 점진적으로 유도시켜 항유방암의 효과의 가능성을 제시하였다.