• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.235-239
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• 2002

It has been reported that lovastatin induced cell death and suppressed proliferation in various cell lines. In this study, we examined whether the cytotoxic effects of lovastatin could be prevented by Bcl-2 or $Bcl-_{XL}$ in C6 glial cells. Overexpression of human Bcl-2 or $Bcl-_{XL}$ prevented lovastatin $(25{\mu}M)-induced$ changes such as DNA fragmentation, chromatin condensation, disruption of cell membrane, and cleavage of poly (ADP-ribose) polymerase. Lovastatin-induced inhibition of cell proliferation was unaffected by Bcl-2 or $Bcl-_{XL}$ overexpression. These results suggest that Bcl-2 and $Bcl-_{XL}$ can prevent lovastatin-induced apoptosis in C6 glial cells, though the inhibition of proliferation remains unaffected by these proteins.

• Archives of Pharmacal Research
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• 제22권5호
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• pp.448-453
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• 1999

In ginsenoside Rh2-treated rat glioma C6Bu-1 cells, apoptotic morphological changes, such as cell shrinkage, chromatin condensation and pyknosis were confirmed by means of electron microscopy. To evaluate whether induction of apoptosis by ginsenoside Rh2 is mediated by the members of Bcl-2 family, we first established C6Bu-1 cells overexpressing Bcl-2. It was demonstrated that the expression of Bcl-2, Bcl-xL, and Bax was not altered in ginsenoside Rh2-treated C6Bu-1 overexpressing C6Bu-1 cells failed to prevent from ginsenoside Rh2-induced cell death. These results suggest the existence of other apoptotic pathway that requires induction of apoptosis by ginsenoside Rh2 rather than the pathway through Bcl-2, $Bcl-x_{L}$ or Bax in C6Bu-1 cells.

• 한국식품영양과학회지
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• 제35권6호
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• pp.671-676
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• 2006

본 연구는 생강의 대표적인 비휘발성 매운맛 성분인 [6]-gingerol이 인체 유방암세포 MDA-MB-231에서 세포사멸에 미치는 영향을 살펴보았으며 그 결과는 다음과 같다. 세포사멸을 억제하는 단백질인 Bcl-2의 발현은 [6]-gingerol의 농도가 증가할수록 발현이 감소되었으며, mRNA 수준에서도 같은 양상을 보였다. 세포사멸을 유도하는 Bax의 단백질 발현은 [6]-gingerol의 농도가 증가되어도 유의적인 차이는 나타나지 않았으며 mRNA 수준에도 별 영향을 미치지 않았다. 그러나 세포사멸의 지표로 사용되는 Bcl-2/Bax의 비율은 [6]-gingerol의 농도가 증가할수록 감소를 보였다. 그리고 [6]-gingerol의 농도가 증가할수록 caspase-3의 활성이 증가하였다. 이상의 결과들로 볼 때, 인체 유방암 세포인 MDA-MB-231에서 [6]-gingerol은 암세포의 증식을 억제하고, 세포사멸을 유도하는 효과가 있는 것으로 사료된다.

• BMB Reports
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• 제54권3호
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• pp.176-181
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• 2021

Bcl-x, a member of the Bcl-2 family, plays a key role in apoptosis. Alternative splicing of Bcl-x pre-mRNA through alternative 5' splice-site selection produces an anti-apoptotic mRNA isoform that includes exon 2b and a pro-apoptotic Bcl-x mRNA isoform that excludes exon 2b. Here we used Bcl-x minigene and identified SRSF2 and SRSF6 as two regulatory factors of 5' splice-site selection of Bcl-x pre-mRNA. We selected binding clusters closer to 5' splice-sites from multiple potential binding sites of SRSF2 and SRSF6 to perform loss of functions analysis through site-directed mutagenesis. Our results demonstrated that these mutations did not abolish regulatory functions of SRSF2 or SRSF6, indicating that a single binding motif or a cluster was not a functional target of these proteins in Bcl-x pre-mRNA splicing. Random deletion mutagenesis did not disrupt the role of SRSF2 and SRSF6. Importantly, mutagenesis of 5' splice-site to a conserved or a weaker score demonstrated that the weaker strength of the target 5' splice-site or higher strength of the other 5' splice-site strength limited the role of SRSF2 and SRSF6 in 5' splice-site activation.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.926-936
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• 2003

Overexpression of human Bcl-2 protein in recombinant Chinese hamster ovary (rCHO) cells producing humanized antibody (SH2-0.32) considerably suppressed sodium butyrate (NaBu)-induced apoptosis during batch culture by using commercially available serum-free medium, which extended the culture longevity. Due to the extended culture longevity provided by the anti-apoptotic effect of Bcl-2 overexpression, the final antibody concentration of 14C6-bcl-2 culture (Bcl-2 high producer, $23\;\mu\textrm{g}\;ml^{-1}$) was 2 times higher than that of the $SH2-0.32-{\Delta}bcl-2$ culture (cells transfected with bcl-2-deficient plasmid, $10.5\;\mu\textrm{g}\;ml^{-1}$) in the presence of NaBu. To determine the effect of NaBu/Bcl-2 overexpression on the molecular integrity of protein products, antibodies purified from 14C6-bcl-2 and $SH2-0.32-{\Delta}bcl-2$ cultures in the presence of NaBu were characterized by using various molecular assay systems. For comparison, antibody purified from the parental rCHO cell culture (SH2-0.32) in the absence of NaBu was also characterized. No significant changes in molecular weight of antibodies could be observed by SDS-PAGE. From GlycoSep-N column analysis, it was found that the core oligosaccharide structure ($GlcNAc_2Man_3GlcNAc_2$) was not affected by NaBu/Bcl-2 overexpression, while the microheterogeneity of N-linked oligosaccharide structure was slightly affected. Compared with the antibody produced in the absence of NaBu, the proportion of neutral oligosaccharides was increased from 10% (14C6-bcl-2) to 16% ($SH2-0.32-{\Delta}bcl-2$) in the presence of NaBu, which was accompanied by the reduced proportion of acidic oligosaccharides, especially of monosialylated and disialylated forms. The changes in microheterogeneous oligoformal structures of antibody in turn affected the mobility of antibody isoforms in isoelectric focusing (IEF), resulting in the occurrence of some more basic antibody isoforms produced in the presence of NaBu. However, the antigen-antibody binding properties were not changed by alteration of glycosylation pattern. The competitive enzyme-linked immunosorbent assay (ELISA) showed that the antibody produced by NaBu/Bcl-2 overexpression maintained its antigen-antibody binding properties with binding affinity of about $2.5{\times}10^9{\;}M^{-1}$. Taken together, no significant effects of NaBu/Bcl-2 overexpression on the molecular integrity of antibodies, produced by using serum-free medium, could be observed by the molecular assay systems.

• Tuberculosis and Respiratory Diseases
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• 제46권1호
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• pp.36-43
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• 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
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• 제16권1호
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• pp.83-86
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• 2015

Background: Diffuse large B-cell lymphoma (DLBCL) is the most common form of non-Hodgkins lymphoma (NHL), accounting for approximately 25% of NHL cases. The aim of this study was to evaluate the association between the BCL6 and MUM1 gene expression and patient prognosis and stage. Materials and Methods: After ethical approval, in a cross-sectional study, tissue samples of 80 patients with diffuse large B-cell lymphoma were analyzed for BCL6 and MUM1 gene expression. Immunohistochemical staining was performed with division into categories of 0-5%, 5-25%, 26-50%, 51-75% and more than 75%. Other clinical and histological information such as lymph node involvement, T-stage, B symptoms and patient outcome were also recorded. Data were analyzed with SPSS version 16 and a P-value less than 0.05 was considered significant. Results: The patient mean age was $46.9{\pm}10.5$ years ($47.6{\pm}10.7$ and $46.1{\pm}9.6$ for males and females, respectively). A significant association was seen between lymphoma stage and BCL6 (p=0.045) but not MUM1 expression (p=0.09). However, the latter was associated with mortality (p=0.006) as was also the BCL6 level (p=0.006). Conclusions: : Overexpression of MUM1 and BCL6 is associated with poor prognosis in patients with diffuse large B-cell lymphoma.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제41회 하계 정기 학술대회 초록집
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• pp.261-261
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• 2011

펄스 직류 전원 $BCl_3/SF_6$ 플라즈마를 이용하여 GaAs/$Al_{0.2}Ga_{0.8}As$의 선택적 식각을 연구하였다. 식각 주요 공정 변수는 $BCl_3/SF_6$ 플라즈마에서 $SF_6$ 가스 유량(0~50%)이었다. $BCl_3/SF_6$의 총 가스 유량은 20 sccm이었다. 다른 공정 조건인 공정 압력(100 mTorr), 펄스 파워(500 V), 펄스 주파수(200 kHz), 리버스 시간 (0.7 ${\mu}s$)은 일정하게 고정시켰으며 기계적 펌프만을 이용하여 공정을 진행하였다. 오실로스코프(Oscilloscope) 데이터에 의하면 가스의 조성 변화에도 척에 걸리는 입력 전압과 전류가 거의 변화가 없었다. $BCl_3/SF_6$ 가스가 10%의 조성에서 GaAs와 $Al_{0.2}Ga_{0.8}As$의 식각 선택비가 약 48 : 1로 우수한 결과를 나타내었다. 그러나 $BCl_3/SF_6$ 가스의 증가는 GaAs의 식각율과 선택도를 감소시켰다. 그리고 $SF_6$ 가스의 조성비가 30% 이상일 경우에는 GaAs와 $Al_{0.2}Ga_{0.8}As$가 식각되지 않았다. 식각 후에 GaAs의 표면 거칠기(RMS surface roughness)는 0.7~1.3 nm로 나타났다. 위의 결과들을 종합적으로 보면 펄스 직류 전원 $BCl_3/SF_6$의 조성비가 10%일 때 가장 좋은 식각 선택비를 얻을 수 있었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.564-570
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• 2005

In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3037-3046
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• 2012

The diffuse large B-cell lymphoma (DLBCL) encompasses two major groups of tumors with uneven survival outcomes - germinal center B-cell (GCB) and non-germinal center B-cell (non-GCB). In the present study, we investigated the expression of GCB markers (BCL-6 and CD10) and non-GCB markers (CD138 and MUM-1) in an effort to evaluate their prognostic value. Paraffin-embedded tumor biopsies of 46 Jordanian DLBCL patients were analyzed, retrospectively, by immunohistochemistry to investigate the expression of BCL-6, CD10, CD138 and MUM-1. In addition, survival curves were calculated with reference to marker expression, age, sex and nodal involvement. Positive expression of BCL-6, CD10, CD138 and MUM-1 was shown in 78%, 61%, 39% and 91% of the cases, respectively, that of BCL-6 being associated with better overall survival (p = 0.02), whereas positive CD138 was linked with poor overall survival (p = 0.01). The expression of CD10 and MUM-1 had no impact on the overall survival. Among the clinical characteristics studied, diagnosis at an early age, nodal involvement and maleness were associated with a higher overall survival for DLBCL patients. Our results underline the importance of BCL-6 as a marker of better prognosis and CD138 as a marker of poor prognosis for DLBCL patients.