• Journal of Microbiology and Biotechnology
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• v.13 no.6
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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.

• The Transactions of The Korean Institute of Electrical Engineers
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• v.56 no.2
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• pp.349-354
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• 2007

In this work, we investigated etching characteristics of $HfO_2$ thin film and Si using inductive coupled plasma(ICP) system. The ion energy distribution functions in an ICP system was analyzed by quadrupole mass spectrometer(QMS) with an electrostatic ion energy analyzer. The maximum etch rate of $HfO_2$ thin film is 85.5 nm/min at a $BCl_3/(BCl_3+Ar)$ of 20 % and decreased with further addition of $BCl_3$ gas. From the QMS measurements, the most dominant positive ion energy distributions(IEDS) showed a maximum at 20 % of $BCl_3$. These tendency was very similar to the etch characteristics. This result agreed with the universal energy dependency of ion enhanced chemical etching yields. And the maximum selectivity of $HfO_2$ over Si is 3.05 at a $O_2$ addition of 2 sccm into the $BCl_3/(BCl_3+Ar)$ of 20 % plasma.

• Journal of Life Science
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• v.19 no.10
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• pp.1489-1494
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• 2009

This study was designed to investigate the effects of swimming and minocycline on motor function recovery and Bcl-2 expression after spinal cord injury (SCI) in rats. After operation, neurological motor behavior test (BBB scale) on days 1, 4, 7, 10, and 14 were tested. Western blot and immunohistochemical assessment (Bcl-2) were performed on day 14. BBB scale started to show a statistically significant difference on day 7 (p<0.05). On day 14, it showed the most significant (p<0.05) difference. Expression of Bcl-2 increased in all the experimental groups. In particular, the highest expression of Bcl-2 appeared in the swimming and minocycline groups. Based on these results, minocycline and swimming were the most effective factors in the motor behavior function and immunohistochemical assessment of SCI rats.

• BMB Reports
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• v.45 no.5
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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.

• ETRI Journal
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• v.30 no.3
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• pp.383-393
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• 2008

The etching mechanism of $ZrO_2$ thin films and etch selectivity over some materials in both $BCl_3$/Ar and $BCl_3/CHF_3$/Ar plasmas are investigated using a combination of experimental and modeling methods. To obtain the data on plasma composition and fluxes of active species, global (0-dimensional) plasma models are developed with Langmuir probe diagnostics data. In $BCl_3$/Ar plasma, changes in gas mixing ratio result in non-linear changes of both densities and fluxes for Cl, $BCl_2$, and ${BCl_2}^+$. In this work, it is shown that the non-monotonic behavior of the $ZrO_2$ etch rate as a function of the $BCl_3$/Ar mixing ratio could be related to the ion-assisted etch mechanism and the ion-flux-limited etch regime. The addition of up to 33% $CHF_3$ to the $BCl_3$-rich $BCl_3$Ar plasma does not influence the $ZrO_2$ etch rate, but it non-monotonically changes the etch rates of both Si and $SiO_2$. The last effect can probably be associated with the corresponding behavior of the F atom density.

• The Journal of Korean Medicine
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• v.22 no.4
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• pp.90-100
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• 2001

Objectives : Bambusae Caulis in Liquamen (BCL) has been used for stamina fortification in Oriental Medicine for thousand years. The goal of the present study was to investigate the muscle antifatigue effect of BCL. Methods : The mice were divided into three groups. The control group was allowed to swim for 10 minutes without BCL infusion. For the preventive effect of BCL, another group referred to BCL-1 was fed with BCL an hour before the swimming exercise. BCL-2 group for the reversion effect was forced to swim for 10 minutes at first, then fed with BCL followed by half an hour rest and another 10 minute swimming exercise. All the serum samples collected by heart puncture were used for the measuring biochemical factors such as LDH, CPK, glucose, lactate, triglycerides and total cholesterol. Results : In the BCL-2 group, lactate and LDH were significantly reduced compared to the control. BCL infusion during the exercise was effective in metabolizing LDH, reducing the conversion ratio of pyruvate into lactate. In the BCL-1 group, it was not effective in antifatigue function. The level of glucose was significantly reduced in BCL-1 group compared to the control. When the BCL was infused before the exercise, it is assumed that glycogenolysis seen during the exercise was prevented. When the BCL was infused during the exercise, the triglycerides and total cholesterol were increased significantly compared to the control. BCL seems to cause the increase of lipolysis to utilize the fat as an energy source. Unlike other parameters, CPK did not show any changes from BCL infusion. Conclusions : Based on the above results, BCL is found to be involved in energy metabolism. Further studies are needed to find out the underlying mechanism of BCL for its effect on stamina fortification.

• Journal of Korean Neurosurgical Society
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• v.40 no.1
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• pp.1-5
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• 2006

Objective : Survivin is an inhibitor of apoptosis protein[IAP], which inhibits apoptosis through a pathway distinct from the Bcl-2 family members. Overexpression of survivin and Bcl-2 have been commonly reported in human neoplasms. The authors investigate whether there is a synergistic effect on the anti-apoptosis rate of primary brain tumors "in situ" based on the co-expression of survivin and Bcl-2. Methods : One hundred and two brain tumor patients who had been resected were included in this study. Survivin tin and Bcl-2 were detected by Western blotting analysis, while apoptosis was examined by DNA fragmentation analysis. An anti-apoptotic rate was assessed in these brain tumor samples based on the expression of survivin and Bcl-2 or co-expression of both. Results : Survivin and Bcl-2 were expressed in 57[55.9%] and 53[52.0%] of 102 brain tumor samples studied respectively, and co-expressed in 31[30.4%]. The percentage of astrocytic and meningeal tumors expressing survivin was significantly correlated with histological grades; however, Bcl-2 was not correlated [p=0.106]. The anti-apoptotic rate in primary brain tumors with survivin, Bcl-2, and both was detected in 49[86.0%] of 57 samples, 42[79.9%] of 53 samples, and 27[87.1%] of 31 samples, respectively. Their difference in the frequency of anti-apoptosis was not significant. Conclusion : Survivin or Bcl-2 is involved in the anti-apoptosis. However, it suggests that co-expression of survivin and Bcl-2, together, have no synergistic effect on the anti-apoptotic properties of the primary brain tumors.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.20
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• pp.8861-8869
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• 2014

Background: The prognostic value of Bcl-2 protein expression in non-small cell lung cancer (NSCLC) is under debate. We therefore systematically reviewed the evidence for Bcl-2 protein effects on NSCLC survival to elucidate this issue. Materials and Methods: An electronic search in Pubmed and Embase complemented by manual searches in article references were conducted to identify eligible studies to evaluate the association between Bcl-2 protein expression and overall survival (OS) as well as disease free survival (DFS) of NSCLC patients. Combined hazard ratios (HRs) with corresponding 95% confidence intervals (95%CIs) were pooled using the random-effects model. Results: A total of 50 trials (including 52 cohorts) encompassing 7,765 patients were pooled in the meta-analysis regarding Bcl-2 expression and OS of NSCLC patients. High expression of Bcl-2 protein had a favorable impact (HR=0.76, 95%CI=0.67-0.86). In the group of Bcl-2 expression and DFS, 11 studies including 2,634 patients were included. The synthesized result indicated high expression of Bcl-2 protein might predict good DFS (HR=0.85, 95%CI=0.75-0.95). Conclusions: Our present meta-analysis demonstrated favorable prognostic values of Bcl-2 expression in patients with NSCLC. Further prospective trails are welcomed to validate the utility of assessing Bcl-2 in NSCLC patient management.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.1
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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.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.7
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• pp.4209-4214
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• 2013

Background: Breast cancer is among the five most common cancers and ranks first among cancers diagnosed in Iranian women. Screening and treatment of this disease with molecular methods, especially regarding high incidences at early age and advanced stage, is essential. Several genes with altered expression have been identified by cDNA microarray studies in breast cancer, with the Bcl-2 gene indicated as a likely candidate. In this study, we studied Bcl-2 gene expression levels in parallel tumor and non-tumor breast tissues. Materials and Methods: Forty samples including 21 tumor, 16 non tumor (marginal) and 3 benign breast tissues which were all pathologically diagnosed, were subjected to RNA extraction and polyA RT-PCR with the expression level of Bcl-2 quantified using real-time PCR. Results: There is higher expression levels of the Bcl-2 gene in tumor samples compared with marginal samples, but not attaining significance(p>0.05). Bcl-2 expression in 14 (66.7%) of the cases of tumor samples and 9 (56.3%) cases of the marginal samples were positive. Comparison of the expression of the Bcl-2 gene in histological grade showed that a high expression of Bcl-2 was associated with a high histological grade (p<0.41). Conclusions: Our data suggests that dysregulated Bcl-2 gene expression is potentially involved in the pathogenesis of breast cancer. Using gene expression analysis may significantly improve our ability for screening cancer patients and will prove a powerful tool in the diagnosis and prognostic evaluation of the disease whilst aiding the cooperative group trials in the Bcl-2 based therapy project.