• Journal of Chest Surgery
• /
• v.41 no.1
• /
• pp.25-33
• /
• 2008

Background: A critical shortage of donor organs has necessitated an investigation of new strategies to increase the availability of additional organs available for human transplantation. We investigated the amount of apoptosis and expression of GADD45 ${\beta}$ in two groups, a GADD45 ${\beta}$-transfected group and untransfected group. Material and Method: The experimental groups consist of a control group (normal H9C2 cell line) and GADD45 ${\beta}$-transfected group. After injury of the each group, we evaluated the expression of GADD45 ${\beta}$ and the level of apoptosis in each group. Result: There was a significant increase in the expression of GADD45 ${\beta}$ in the GADD45 ${\beta}$-transfected group at 1 hour, 2 hours, and 3 hours after stimuli as compared with the control group. The amount of cardiac myoblast cell line apoptosis was significantly lower in the GADD45 ${\beta}$-transfected group as compared with the control group. The concentration of annex in in the GADD45 ${\beta}$-transfected group was significantly lower than that of the control. group after cell. injury. Conclusion: Transfection of a rat myoblast cell line with the GADD45 ${\beta}$ gene results in. decreased susceptibility to cell injury of human serum.

• Journal of Life Science
• /
• v.18 no.4
• /
• pp.461-466
• /
• 2008

Transforming growth $factor-{\beta}$ ($TGF-{\beta}$)-dependent apoptosis is important in the elimination of damaged or abnormal cells from normal tissues in vivo. Gadd45b has been known to participate in $TGF-{\beta}-induced$ apoptosis by the activation of p38 kinase. In this report, we show that Gadd45b is an immediate-early response gene for $TGF-{\beta}$ during apoptosis in EpH4 cells. To elucidate the molecular mechanism of $TGF-{\beta}-induced$ Gadd45b gene expression, we cloned the 5'-flanking region of the mouse Gadd45b gene. When transfected into EpH4 cells, this 5'-flanking region conferred promoter activity and inducibility by $TGF-{\beta}$. Deletion analyses demonstrated that the minimal promoter activity was detected in the proximal region 220 bp upstream of the transcription initiation site. We also found that the proximal Gadd45b promoter is activated by $TGF-{\beta}$ through the action of Smad2, Smad3, and Smad4. Finally, we show that the expression of Gadd45b gene by $TGF-{\beta}$ is suppressed in EpRas cells in which $TGF-{\beta}$ could not induce apoptosis, suggesting that Gadd45b may be a crucial target for $TGF-{\beta}-induced$ apoptosis in EpH4 cells.

• Journal of Life Science
• /
• v.21 no.5
• /
• pp.753-760
• /
• 2011

In the present study, the effects of paprika extract and its components including vitamin C, lycopene and beta-carotene on cell death in ultraviolet B (UVB)-exposed HaCaT cells were investigated. The cell viability upon treatment for 24 hr with either paprika extract or vitamin C alone was similar to or greater than that of the untreated control. However, the viability of the cells treated with lycopene or beta-carotene decreased to about 20% of that in the untreated control. When UVB-exposed cells were post-incubated for 24 hr in medium containing paprika extract or vitamin C, cell viability increased in a concentration dependent manner as compared to those post-incubated in a normal growth medium. In contrast, post-incubation of UVB-exposed cells with lycopene or beta-carotene decreased cell viability in a concentration dependent manner as compared to those post-incubated in a normal growth medium. The nuclear fragmentation analysis showed that paprika extract or vitamin C decreases UVB-induced apoptosis. The apoptotic nuclear fragmentation resulting from UVB exposure was also protected by the paprika extract or vitamin C post-incubation. However, the UVB-induced apoptotic nuclear fragmentation of the cells treated with lycopene or beta-carotene increased in a concentration dependent manner. Western blot analysis showed that either paprika extract or vitamin C treatment alone did not significantly change the level of p53 and GADD45 protein. Interestingly, post-incubation of UVB-exposed cells with paprika extract or vitamin C decreased the p53 and GADD45 protein level as compared to those post-incubated in a normal growth medium. In contrast, incubation of UVB-exposed or non-irradiated cells with lycopene or beta-carotene increased the p53 and GADD45 protein levels in a concentration dependent manner as compared to those incubated in a normal growth medium. All these results suggest that paprika extract and vitamin C help the survival of the UVB-exposed cells, while lycopene and beta-carotene potentiate the apoptotic death of UVB-exposed cells, in accordance with the respective changes in p53 and GADD45 protein levels.

• Molecules and Cells
• /
• v.40 no.12
• /
• pp.916-924
• /
• 2017

MicroRNAs are widely involved in the pathogenesis of cardiovascular diseases through regulating gene expression via translational inhibition or degradation of their target mRNAs. Recent studies have indicated a critical role of microRNA-206 in myocardial ischaemia-reperfusion (I/R) injury. However, the function of miR-206 in myocardial I/R injury is currently unclear. The present study was aimed to identify the specific role of miR-206 in myocardial I/R injury and explore the underlying molecular mechanism. Our results revealed that the expression level of miR-206 was significantly decreased both in rat I/R group and H9c2 cells subjected to hypoxia/reoxygenation (H/R) compared with the corresponding control. Overexpression of miR-206 observably decreased infarct size and inhibited the cardiomyocyte apoptosis induced by I/R injury. Furthermore, bioinformatics analysis, luciferase activity and western blot assay proved that $Gadd45{\beta}$ (growth arrest DNA damage-inducible gene $45{\beta}$) was a direct target gene of miR-206. In addition, the expression of pro-apoptotic-related genes, such as p53, Bax and cleaved caspase3, was decreased in association with the down-regulation of $Gadd45{\beta}$. In summary, this study demonstrates that miR-206 could protect against myocardial I/R injury by targeting $Gadd45{\beta}$.

• Journal of Life Science
• /
• v.24 no.2
• /
• pp.112-117
• /
• 2014

We investigated the effects of extracts from Rubus coreanus (RC) and Artemisia princeps var. orientalis (AP) on DNA damage response in ultraviolet B (UVB)-exposed HaCaT cells. Cell activity upon treatment for 24 h with RC or AP alone was similar to or greater than that of the nontreated control. When UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, cell activity increased in a concentration-dependent manner. Nuclear fragmentation analysis showed that postincubation with RC or AP decreased UVB-induced apoptosis by about 20% and 15%, respectively, of that in cells postincubated with growth medium. When UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, the level of cyclobutane pyrimidine dimer decreased in a concentration- dependent manner. Western blot analysis showed that treatment of cells not exposed to UVB with RC or AP alone did not significantly change the levels of phospho-p53 and GADD45 protein. Interestingly, when UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, phospho-p53 and GADD45 levels decreased in a concentration dependent manner. Our results suggest that RC and AP extract assist the survival of UVB-exposed cells in parallel with a decrease in levels of UVB-induced DNA damage and damage-response proteins, such as p53 and GADD45.

• Toxicological Research
• /
• v.25 no.3
• /
• pp.119-124
• /
• 2009

Genes related to Mycoplasma hyopneumoniae-induced inflammation were identified using the genefishing technology, an improved method for identifying differentially expressed genes (DEGs) using an annealing control primer (ACP) system in RAW264.7 cells. After treatment with M. hyopneumoniae, 16 DEGs were expressed in RAW264.7 cells using a pre-screening system. Among these 16 DEGs, 11 DEGs (DEGs 1, 4, 5-10, 12-15) were selected and sequenced directly, revealing that DEG12 (Grap), DEG14 (Gadd45), and DEG15 (secreted phosphoprotein 1) were related to inflammatory cytokines. This is the first report that intact M. hyopneumoniae induces the expression of Grap, Gadd 45${\beta}$, and secreted phosphoprotein 1 in RAW264.7 cells. Subsequently, these genes may be targets for screening novel inhibitors of the mycoplasmal inflammatory response.

• Proceedings of the Korea Environmental Mutagen Society Conference
• /
• 2004.11a
• /
• pp.139-139
• /
• 2004

• Proceedings of the Korean Society of Life Science Conference
• /
• 2005.04a
• /
• pp.47-51
• /
• 2005

• The Journal of Korean Medicine
• /
• v.20 no.1 s.37
• /
• pp.91-105
• /
• 1999

The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

• Journal of Ginseng Research
• /
• v.34 no.3
• /
• pp.205-211
• /
• 2010

Korean red ginseng (KRG) has been used worldwide as a traditional medicine for the treatment of various diseases, including cancer. In this study, we determined the effect of KRG on the responses of HaCaT cells to peroxynitrite ($ONOO^-$). Cells has been used worldwide as a traditional medicine for the treatment of various diseases, including cancer. In this study, we determined the effect of KRG on the responses of HaCaT cells to peroxynitrite ($ONOO^-$). Cells treated with $ONOO^-$ (2 mM) prior to incubation with control medium for 12 hours displayed reduced viability, as determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (viability about 48% of that of non-treated control cells). When KRG was added to the post-incubation medium, the negative effects of $ONOO^-$ on cell viability were significantly reduced. Reverse transcription-polymerase chain reaction analysis indicated that KRG alone did not significantly alter p53 or "growth arrest and DNA damage" (GADD)45 mRNA levels. However, the addition of KRG to the post-incubation medium significantly and dose-dependently reduced levels of p53 and GADD45 mRNA in $ONOO^-$-treated cells. Western blot analyses revealed that incubation with KRG decreased p53 and GADD45 protein levels in $ONOO^-$-treated cells, relative to those in cells incubated with control medium. Collectively, these results suggest that Korean red ginseng extract protects cells against $ONOO^-$-induced genotoxicity by increasing cell viability through modulating the expression of p53 signaling intermediates.