• Tuberculosis and Respiratory Diseases
• /
• 제46권1호
• /
• pp.36-43
• /
• 1999

연구배경: bcl-2는 apoptosis를 억제하여 세포 수명을 연장시켜 종양세포의 생존기간 중 다른 암유전자의 추가척인 영향을 받을 수 있는 기회를 증가시켜 암을 유발하는 암유전자이다. 이러한 bcl-2는 apoptosis 억제에도 불구하고, 임상적으로 bcl-2 양성인 경우에 그 기전은 아직 확실치 않으나 오히려 예후가 양호하다는 보고가 있다. 이에 저자들은 비소세포 폐암에서 bcl-2의 발현률을 계측하고, 세포분열주기와 비교한 후 생존률과의 관계를 검색하였다. 방 법: 원발성 비소세포 폐암으로 확진받고, 외과적 절제술후 paraffin에 보관된 57례의 병리조직에서 면역조직화학 염색법으로 bcl-2의 발현을 확인하고, bcl-2와 병리조직형, TNM 병기, 유식세포분석법에 의한 세포주기비율(S-and $G_1$-phase fraction) 그리고 생존기간과의 관계를 분석하였다. 결 과: 57례 중 남녀비는 43:14 였고, 조직학적 분류는 편평상피암 34례, 선암 19례, 대세포암 4례였으며, TNM I 병기 14례, II 병기 21례, III 병기 22례였다. bcl-2는 43.8% (25/57)에서 발현되었으며 면평상피암 47%(16/34), 선암 32%(6/19)였으나, 유의한 차이는 없었고, TNM I 병기 28.6%(4/14), II 병기 52.3% (11/21), III 병기 45.5%(10/22)로서 역시 유의한 차이는 없었다. bcl-2 발현군과 비발현군에서의 $G_1$-주기비율은 75.5($\pm10.8$)%, 65.5($\pm11.4$)% 로서 유의한 차이가 있었으며 (p<0.05), S-주기비율은 14.1($\pm7.8$)%, 24.7($\pm10.5$)%로서 역시 유의하게 bcl-2 발현군의 세포분열능이 저하되어 있었다 (p<0.005). bcl-2 발현군과 비발현군에서의 2년 생존률은 65%, 71%, 3년 생존률은 54%, 52%, 5년 생존률은 41%, 46% 그리고 중간 생존기간은 53개월, 37개월이었으나 통계적 유의성에 이르지는 못하였다. (p>0.05, Kaplan-Meier, log rank) 결 론: 비소세포 폐암에서 bcl-2는 43.8%에서 발현되었으며, 병리조직형, TNM 병기에 따른 발현률의 차이는 없었다. bcl-2 발현군은 비발현군에 비하여 S-주기비율은 유의하게 감소하고, $G_1$-주기비율은 유의하게 증가되었다. bcl-2 발현군의 중간 생존기간은 53개월로서 비발현군의 37개월보다 연장되었으나 통계적 유의성에 이르지는 못하였다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권4호
• /
• pp.2387-2391
• /
• 2013

Resistance to apoptosis is a major obstacle preventing effective therapy for malignancies. Bcl-2 plays a significant role in inhibiting apoptosis. We reconstructed a stable human Bcl-2 transfected cell line, BIU87-Bcl-2, that was derived from the transfection of human bladder carcinoma cell line BIU87 with a plasmid vector containing recombinant Bcl-2 [pcDNA3.1(+)-Bcl-2]. A cell line transfected with the plasmid alone [pcDNA3.1(+)-neo] was also established as a control. BIU87 and BIU87-neo proved sensitive to adriamycin induced apoptosis, while BIU87-Bcl-2 was more resistant. In view of the growing evidence that NF-${\kappa}B$ may play an important role in regulating apoptosis, we determined whether Bcl-2 could modulate the activity of NF-${\kappa}B$ in bladder carcinoma cells. Stimulation of BIU87, BIU87-neo and BIU87-Bcl-2 with ADR resulted in an increase expression of NF-${\kappa}B$ (p<0.001). The expression of NF-${\kappa}B$ in BIU87-Bcl-2 was higher than in the other two cases, with a concomitant reduction in the $I{\kappa}B{\kappa}$ protein level. These results suggest that the overexpression of Bcl-2 renders human bladder carcinoma cells resistant to adriamycin-induced cytotoxicity and there is a link between Bcl-2 and the NF-${\kappa}B$ signaling pathway in the suppression of apoptosis.

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

• 한국미생물·생명공학회지
• /
• 제50권2호
• /
• pp.293-300
• /
• 2022

Lipoteichoic acid (LTA) regulates the immune system, including inflammatory responses, through TLR2-mediated signaling pathways. LTA isolated from Staphylococcus aureus (aLTA) has been shown to induce apoptosis, but the detailed mechanism has not been identified. We found that aLTA induced apoptosis through an autocrine mechanism in the human monocyte-like cell line, THP-1. We observed that the expression level of the anti-apoptosis protein, Bcl2, was suppressed in LTA-treated THP-1 cells. In addition, the cytokines, TNF-α and IFN-γ, which have been shown to induce apoptosis in some cell lines, were involved in THP-1 cell death via the modulation of Bcl2. The suppression of Bcl2 by aLTA was recovered when the negative regulator, SOCS-1, was knocked down. Taken together, these results showed that aLTA induced apoptosis in THP-1 cells through an autocrine mechanism of cytokines and SOCS-1-mediated Bcl2 inhibition.

• Toxicological Research
• /
• 제18권4호
• /
• pp.411-416
• /
• 2002

Apoptosis, or programmed cell death, is a genetically regulated pathway that is altered in many cancers. Overexpression of bcl-2 leads to resistance to apoptosis and promotes tumorigenesis. To determine the effect of bcl-2 antisense treatment in human lung cancer cell lines, a 20 mer full phosphorothioate oligonucleotide (ODN) targeted at the coding region of the bcl-2 mRNA was synthesized. Western blot analyses were used to examine bcl-2 protein level in five human non-small cell lung cancer (NSCLC) cell lines (NCI-H226, SK-MES-1 NCI-H358, NCI-H522 and NCI-Hl 299) and four human small cell lung cancer (SCLC) cell lines (NCI-H69, NCI-H4l7, HCC-2108 and SW2). Three out of five NSCLC (NCI-H226, SK-MES-1 and NCI-Hl 299) and all of SCLC cell lines expressed Bcl-2 protein. Treatment of these cell with antisense ODN for 48 hours reduced their viability and Bcl-2 protein level. As a conclusion, bcl-2 antisense treatment appears reduction of the Bcl-2 protein levels and cytotoxic effect including apoptosis in human lung cancer cell lines.

• Journal of Chest Surgery
• /
• 제31권12호
• /
• pp.1200-1205
• /
• 1998

연구목적: 폐암은 protooncogene의 활성화와 종양억제유전인자(tumor suppressor gene)의 비활성화 등 다단 과정에 의하여 발생한다. 치료목적의 완전 절제가 가능하였던 제 IIIA기 비소세포폐암 환자에서 cyclin D1, p53, bcl-2 gene의 변이가 폐암에 미치는 영향을 조사하고자 하였다. 대상 및 방법: 1990년부터 1995년까지 연세의료원에서 치료목적의 완전절제가 가능하였던 stageIIIA 비소세포폐암 환자 100명의 paraffin block과 임상기록을 이용하였다. 각 환자의 조직절편을 labelled streptavidin-biotin method로 immunohistochemical 염색하였고 cyclin D1, p53, Bcl-2 immunostaining을 위한 조직절편들은 immunostaining하기 전 citrate buffer 내에서 10분에서 20분간 microwave oven으로 전처치한 후 cyclin D1은 NCL-cyclin D1-GM으로 p53는 lone DO-7으로 bcl-2는 clone 124로 overnight incubation하였다. 수술 후 평균 추적조사기간은 24.1 개월(range; 2∼84 개월)이었다. 결과: 100예의 폐암 중 56예가 편평상피세포암, 37예가 선암, 5예가 adenosquamous cell carcinoma, 2예가 대세포암이었고 수술 후 5년 생존율은 32.1%이었다. cyclin D1의 양성율은 35 %, p53는 56 %, bcl-2는 17 %였으나 cyclin D1, p53, Bcl-2 단백질 양성 발현과 생존율과의 상관관계는 없었다. 결론: 연구결과 cyclin D1, p53, Bcl-2 단백질 양성 발현이 비소세포폐암 발생기전과 연관되어 있으나 수술 후 예후인자로서는 부적당한 것으로 판단되었다.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 2002년도 창립10주년기념 및 국립독성연구원 의약품동등성평가부서 신설기념 국재학술대회:생물학적 동등성과 의약품 개발 전략을 위한 국제심포지움
• /
• pp.113-113
• /
• 2002

Phylogenetically conserved Bcl-2 family proteins play a pivotal role in the regulation of apoptosis from virus to human. Members of the Bcl-2 family consist of antiapoptotic proteins such as Bcl-2, Bcl-xL, and Bcl-w, and proapoptotic proteins such as BAD, Bax, BOD, and Bok. It has been proposed that anti- and proapoptotic Bcl-2 proteins regulate cell death by binding to each other and forming heterodimers. A delicate balance between anti- and proapoptotic Bcl-2 family members exists in each cell and the relative concentration of these two groups of proteins determines whether the cell survives or undergoes apoptosis. Mcl-1 (Myeloid cell :leukemia-1) is a member of the Bcl-2 family proteins and was originally cloned as a differentiation-induced early gene that was activated in the human myeloblastic leukemia cell line, ML-1 . Mcl-1 is expressed in a wide variety of tissues and cells including neoplastic ones. We recently identified a short splicing variant of Mcl-1 short (Mcl-IS) and designated the known Mcl-1 as Mcl-1 long (Mcl-lL). Mcl-lL protein exhibits antiapoptotic activity and possesses the BH (Bcl-2 homology) 1, BH2, BH3, and transmembrane (TM) domains found in related Bcl-2 proteins. In contrast, Mcl-1 S is a BH3 domain-only proapoptotic protein that heterodimerizes with Mcl-lL. Although both Mc1-lL and Mcl-lS proteins contain BH domains fecund in other Bcl-2 family proteins, they are distinguished by their unusually long N-terminal sequences containing PEST (proline, glutamic acid, serine, and threonine) motifs, four pairs of arginine residues, and alanine- and glycine-rich regions. In addition, the expression pattern of Mcl-1 protein is different from that of Bcl-2 suggesting a unique role (or Mcl-1 in apoptosis regulation. Tankyrasel (TRF1-interacting, ankyrin-related ADP-related polymerasel) was originally isolated based on its binding to TRF 1 (telomeric repeat binding factor-1) and contains the sterile alpha motif (SAM) module, 24 ankyrin (ANK) repeats, and the catalytic domain of poly(adenosine diphosphate-ribose) polymerase (PARP). Previous studies showed that tankyrasel promotes telomere elongation in human cells presumably by inhibiting TRFI though its poly(ADP-ribosyl)action by tankyrasel . In addition, tankyrasel poly(ADP-ribosyl)ates Insulin-responsive amino peptidase (IRAP), a resident protein of GLUT4 vesicles, and insulin stimulates the PARP activity of tankyrase1 through its phosphorylation by mitogen-activated protein kinase (MAPK). ADP-ribosylation is a posttranslational modification that usually results in a loss of protein activity presumably by enhancing protein turnover. However, little information is available regarding the physiological function(s) of tankyrase1 other than as a PARP enzyme. In the present study, we found tankyrasel as a specific-binding protein of Mcl-1 Overexpression of tankyrasel led to the inhibition of both the apoptotic activity of Mel-lS and the survival action of Mcl-lL in mammalian cells. Unlike other known tankyrasel-interacting proteins, tankyrasel did not poly(ADP-ribosyl)ate either of the Mcl-1 proteins despite its ability to decrease Mcl-1 proteins expression following coexpression. Therefore, this study provides a novel mechanism to regulate Mcl-1-modulated apoptosis in which tankyrasel downregulates the expression of Mcl-1 proteins without the involvement of its ADP-ribosylation activity.

• 생명과학회지
• /
• 제13권1호
• /
• pp.118-127
• /
• 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.

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

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권2호
• /
• pp.895-901
• /
• 2013

Resistance to induction of apoptosis is a major obstacle for bladder cancer treatment. Bcl-2 is thought to be involved in anti-apoptotic signaling. In this study, we investigated the effect of Bcl-2 overexpression on apoptotic resistance and intracellular reactive oxygen species (ROS) generation in bladder cancer cells. A stable Bcl-2 overexpression cell line, BIU87-Bcl-2, was constructed from human bladder cancer cell line BIU87 by transfecting recombinant Bcl-2 [pcDNA3.1(+)-Bcl-2]. The sensitivity of transfected cells to adriamycin (ADR) was assessed by MTT assay. Apoptosis was examined by flow cytometry and acridine orange fluorescence staining. Intracellular ROS was determined using flow cytometry, and the activities of superoxide dismutase (SOD) and catalase (CAT) were also investigated by the xanthinoxidase and visible radiation methods using SOD and CAT detection kits. The susceptibility of BIU87-Bcl-2 cells to ADR treatment was significantly decreased as compared with control BIU87 cells. Enhanced expression of Bcl-2 inhibited intracellular ROS generation following ADR treatment. Moreover, the suppression of SOD and CAT activity induced by ADR treatment was blocked in the BIU87-Bcl-2 case but not in their parental cells. The overexpression of Bcl-2 renders human bladder cancer cells resistant to ADR-induced apoptosis and ROS might act as an important secondary messenger in this process.