• Biotechnology and Bioprocess Engineering:BBE
• /
• v.8 no.2
• /
• pp.76-82
• /
• 2003

An increased understanding of apoptosis makes anti-apoptosis engineering possible, which is an approach used to inhibit apoptosis for the purpose of therapeutic, or industrial applications in the treatment of the diseases associated with increased apoptosis, or to improve the productivity of animal cell cultures, respectively. Some known anti-apoptosis proteins are the Bcl-2 family, IAP (inhibitor of apoptosis) and Hsps (heat shock proteins), with which anti-apoptosis engineering has progressed. This article reviews anti-apoptosis engineering using known anti-apoptosis compounds, and introduces a 30 K protein, isolated from silkworm hemolymph, as a novel anti-apoptotic protein, that Shows no homology with other known anti-apoptotic proteins. The regulation of apoptosis, using anti-apoptotic proteins and genes originating from the silkworm, Bombyx mori, may provide a new strategy in this field.

• The Journal of Internal Korean Medicine
• /
• v.35 no.1
• /
• pp.79-91
• /
• 2014

Objectives : To investigate the anti-cancer effect of Palbohoichoon-tang (PBHCT) extracts. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were microscopically analyzed after staining with $10{\mu}M$ 2-[4-amidinophenyl]-6-indolecarbamidine dihydrochloride (DAPI) and TUNEL. We also analyzed expression of Bcl2, $Bcl_{xL}$, Bax, procaspase-3, procaspase-9, and procyclic acidic repetitive protein (PARP) by western blot method. Results : Observations showed that PBHCT induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. Western blot analysis of total cell lysates revealed that the PBHCT induced cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP). In addition, PBHCT dose-dependently increased the activity of caspase-9, caspase-3 and PARP-1. Furthermore, PBHCT reduced anti-apoptotic Bcl2, $Bcl_{xL}$ expression which contributed to the loss of mitochondrial membrane potential and the activations of caspase-9 and caspase-3. Conclusions : These findings suggest that PBHCT exerts anti-cancer effects on human neuroblastoma SH-SY5Y cells by inducing apoptotic death via down-regulation of anti-apoptotic proteins such as Bcl2 and $Bcl_{xL}$, up-regulation of pro-apoptotic proteins such as Bax, and activation of caspase cascades and PARP-1.

• Toxicological Research
• /
• v.22 no.1
• /
• pp.31-37
• /
• 2006

It was previously found that melittin inhibited $NF-{\kappa}B$ activity by reacting with signal molecules of $NF-{\kappa}B$ which is critical contributor in cancer cell growth by induction of apoptotic cell death. We here investigated whether melittin inhibits cell growth of human prostate cancer cells through induction of apoptotic cell death, and the possible signal pathways. Melittin ($0{\sim}1\;{\mu}g/ml$) inhibited prostate cancer cell growth in a dose dependent manner. Conversely related to the growth inhibitory effect, melittin increased the induction of apoptotic cell death in a dose dependent manner. Melittin also inhibited DNA binding activity of $NF-{\kappa}B$, an anti-apoptotic transcriptional factor. Consistent with the induction of apoptotic cell death and inhibition of $NF-{\kappa}B$, melittin increased the expression of pro-apoptotic proteins caspase-3, and Bax but down-regulated anti-apoptotic protein Bcl-2. These findings suggest that melittin could inhibit prostate cancer cell growth, and this effect may be related with the induction of apoptotic cell death via inactivation of $NF-{\kappa}B$.

• Korean Journal of Plant Resources
• /
• v.31 no.1
• /
• pp.16-23
• /
• 2018

Paeonia japonica is a perennial flowering plant used in traditional medicine therapy. The purpose of this study was to investigate the effect of water extract and solvent fractions obtained from P. japonica on anti-oxidative, anti-thrombin, anti-invasive and pro-apoptotic activities in YD-10B cells, human oral squamous carcinoma cell line. Water fraction revealed the highest extraction yield at 11.44% (w/w). Anti-oxidative activity was the highest in ethyl acetate fraction (85.13%). In the thrombin inhibitory activity test, ethyl fraction was the highest, with a value of 87.54%. Release and activation of MMP-2/pro-MMP-2 ratio in thrombin-treated YD-10B cells were significantly inhibited in the ethyl acetate fraction. At a concentration of $120{\mu}g/m{\ell}$ water extract and solvent fractions of P. japonica inhibited cell proliferation in YD-10B cells except water fraction. Pro-apoptotic effect on human oral squamous carcinoma cell using the Bax/Bcl-2 ratio analysis was higher in water extract than other fractions. These findings suggest that the ethyl acetate fraction of P. japonica potentiates a promising antioxidant, anti-thrombin and anti-invasive agents.

• The Journal of Internal Korean Medicine
• /
• v.29 no.1
• /
• pp.42-66
• /
• 2008

Aconiti Tuber is a traditional medicinal plant generally used in Oriental medicine therapy. In this study, we investigated the biochemical mechanisms of anti-proliferative effects by the methanol extract of Aconiti tuber (MEBJ) in Caki-1 human renal cell carcinoma cells. It was found that MEBJ could inhibit, in a dose-dependent manner, cell growth which was associated with apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of apoptotic-sub G1 phase. Apoptosis of Caki-1 cells by MEBJ was associated with an up-regulation of pro-apoptotic Bax expression, and a down-regulation of anti-apoptotic Bcl-2 in a dose-dependent manner; however, the levels of IAP family were not affected. MEBJ treatment also induced the proteolytic activation of caspase-3 and -8, and a inhibition of poly(ADP-ribose) polymerase (PARP) and $PLC{\gamma}1$ protein. Furthermore, MEBJ treatment caused a dose-dependent inhibition of iNOS and cyclooxygenase-2 (Cox-2). Though further studies will be needed to identify the active compounds that confer the anti-cancer activity of MEBJ, the present findings provide important new insights into the possible molecular mechanisms of the apoptotic activity of MEBJ in cancer cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.27 no.6
• /
• pp.511-518
• /
• 2001

The effect of mycolactone, a recently reported apoptosis-inducing factor, was investigated in SCC15 oral squamous cell carcinoma(OSCC) cell line. Mycolactone rapidly induced cell death in OSCC cells in 2days, which was similar to that found in apoptotic cell such as detaching from culture plate and rounding-up of cells. Apoptotic cells were increased 4hrs after mycolactone treatment and more than half of cells showed apoptosis after 72hrs. Caspase 3 activation a biochemical evidence of apoptosis, was determined by Western blotting. Caspase 3 activation was started at 2hrs that lasted until 8hrs after mycolactone treatment. The expression of bcl-2 family genes was determined to explain the mechanism of apoptosis found in OSCC cells. The expressions of bad, bak, and bax (pro-apoptotic genes) and bcl-w and bcl-2 genes (anti-apoptotic genes) were not changed by mycolactone treatment. The expression of bcl-xi was decreased 8 hrs after mycolactone treatment. Mcl-1 expression was initially increased at 2 hrs which was decreased 8 hrs after mycolactone treatment. The down-regulation of these two anti-apoptotic genes might explain the mycolactone-induced apoptosis in OSCC cells. In this study, mycholactone was revealed to induce cell death in OSCC cells apoptosis and the apoptosis mechanism of OSCC cells was shown to be down-regulation of anti-apoptotic genes, bcl-xi and mcl-1. These results suggested the applicability of mycolactone for the development of an anti-cancer drug candidate by inducing apoptosis of OSCC cancer cell.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.21
• /
• pp.9445-9451
• /
• 2014

Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK-N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

• The Korea Journal of Herbology
• /
• v.27 no.4
• /
• pp.45-51
• /
• 2012

Objectives : WHW is a polyherbal medicine for the treatment of chronic renal failure (CRF). WHW previously reported various biological property such as anti-inflammation, anti-oxidation and anti-renal fibrosis in CRF. This study aimed to investigate the anti-apoptotic effect of WHW on staurosporin(SSP)-induced apoptosis in canine kidney epithelial cells (MDCK). Methods : MDCK cells were treated with different concentrations of WHW (0.1, 0.2, 0.5 and $1mg/m{\ell}$) for 1 h, and then induced apoptosis by treatment of SSP ($1{\mu}M$) for 24 h. Cell viability was measured by WST-1 assay. The expression of apoptotic proteins such as caspase-3, Bax and Bcl-2 was determined by Western blot. Caspase-3 activity and ROS levels were also measured by their commercial available assay kits. Cell apoptosis was observed by Hoechst and DNA fragmentation. Results : WHW significantly increased the cell viability on SSP-treated MDCK cells. WHW inhibited SSP-induced expression of apoptotic proteins such as caspase-3 and Bax, and significantly decreased caspase-3 activity in MDCK cells. WHW significantly decreased SSP-induced production of ROS, and suppressed SSP-induced chromatin condensation and DNA fragmentation in MDCK cells. Conclusions : These results suggest that WHW has an anti-apoptotic effect in renal cells through suppressing the expression of apoptotic proteins, ROS production and DNA damages.

• 한국생물공학회:학술대회논문집
• /
• 2005.04a
• /
• pp.210-213
• /
• 2005

The suppression of apoptosis during the cell culture might increase recombinant protein production. In the present study, the effects of anti-apoptotic agents on the apoptosis of recombinant Chinese Hamster Ovary cells and the production of Iduronate 2-sulphatase(IDS) were investigated Cell density slightly increased when $2{\mu}M$ of EGCG and $10{\mu}g/mL$ of STR-G were added to culture medium after two days. It was observed that the percentage of apoptotic cells was decreased in the culture with STR-G, and Bcl-2 expression level was enhanced in both culture with STR-G and EGCG. These results suggest that G418 and EGCG are effective anti-apoptotic agents for increasing the productivity of IDS with recombinant CHO-DG44.

• Molecules and Cells
• /
• v.28 no.6
• /
• pp.509-513
• /
• 2009

Here, we show that JNK1 and JNK3 have different roles in ${\alpha}-$ or etoposide-induced apoptosis in HeLa cells. Dominant negative JNK1 inhibited $TNF-{\alpha}-$ or etoposide-induced apoptosis, while dominant negative JNK3 promoted $TNF-{\alpha}-$ or etoposide-induced apoptosis. During $TNF-{\alpha}$-induced apoptosis, JNK1 was activated in a biphasic manner, exhibiting both transient and sustained activity, whereas JNK3 was activated early and in a transient manner. The role of JNK3 activation was an anti-apoptotic effect, while the role of JNK1 activation was a pro-apoptotic effect. These results suggest that the anti-apoptotic mechanism of JNK3 in $TNF-{\alpha}$-induced apoptosis originates before the apoptotic machinery is triggered.