• Asian Pacific Journal of Cancer Prevention
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• 제17권sup3호
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• pp.293-297
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• 2016

TRAIL, tumor necrosis factor (TNF)-related apoptosis-inducing ligand belongs to one of important cytokine superfamilIES, tumor necrosis factor ($TNF{\alpha}$). TRAIL-2 receptor agonists activate several cell signaling pathways in cells in different manners and could lead to apoptosis or necrosis. Agonistic egg yolk antibodies like IgY which have been developed in a selective manner could activate TRAIL death receptors such as TRAIL-2 (DR5) and thus apoptosis signaling. We here investigated induction of apoptosis in human breast cancer cells (MCF7 cell line) by an IgY produced against an 21 aminoacid epitope of the human TRAIL-2 receptor. As the first step a small peptide of 21 aminoacids choosen from the extracellular domain of DR5 protein was produced with a peptide synthesizer. After control assays and confirmation of the correct amino acid sequence, it was injected to hens immunized to achieve high affinity IgYs. At the next step, the produced IgYs were extracted and examined for specificity against DR5 protein by ELISA assay. Subsequently, the anticancer effect of such IgYs was determined by MTT assay in the MCF7 human breast cancer cell line. The produced peptides successfully immunized hens and the produced antibodies which accumulated in egg yolk specifically recognized the DR5 protein. IgYs exerted significant toxicity and killed MCF7 cells as shown by MTT assay.

• 한국미생물·생명공학회지
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• 제24권1호
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• pp.126-131
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• 1996

HL-60 was cultivated to produce tPA (tissue-type plasminogen activator) and study the mechanism of cell death. Maximum cell density and tPA production were obtained as $5.27{\times}10^6$ cells/ml and 324ng/ml, respectively under perfusion cultivation. tPA production was enhanced to 420ng/ml in adding 160nM of phorbol ester. The cells were gradually differentiated to granulocytes rather than proliferation. By Fluorescent microscope, apoptosis was prevailed except the death phase and in high agitation speed, but necrosis was prevailed in thawed cells and during the latter periods of the cultivation. It was also proved that tPA was most produced in apoptosis. To obtain higher tPA productivity, the cells must be maintained in apoptosis, not necrosis phase when the cells were dying.

• 대한핵의학회지
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• 제38권2호
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• pp.190-197
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• 2004

Apoptosis plays a role in the pathophysiology of many kinds of diseases and in the response of treatment. Compared to the necrosis, the apoptosis is a genetically controlled and energy-dependent process which removes the unwanted cells from the body; programmed cell death or cell suicide. During the apoptosis, phosphatidylserine is expressed in the cytoplasmic outer membrane in the early phase. Annexin V, an endogenous human protein (MW=35 kD), has an affinity of about $10^{-9}\;M$ for the phosphatidylserine exposed on the outer membrane of apoptotic cells. Annexin V can be radiolabeled with $^{99m}Tc$ by HYNIC or EC chelators, which can be used as an radiotracer for the in vivo imaging of apoptosis. In this article, we reviewed the apoptosis, radiolabeling of annexin V, and the experimental and clinical data using annexin V imaging.

• Toxicological Research
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• 제22권4호
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• pp.329-332
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• 2006

Ethanolic extract of Aloe vera (Aloe barbadensis Miller) was examined for the cellular toxicity on HepG2 human hepatocellular carcinoma cells. Treatment with Aloe vera extract resulted in DNA fragmentation but not LDH release, suggesting an apoptosis instead of necrosis. Aloe vera induced cytotoxicity was mediated by decrease in ATP levels, whereas GSH depletion, increase in intracellular $Ca^{2+}$, or activation of caspase-3/7 could not be observed with statistical significance. No activation of caspase-3/7 suggests the possibility of caspase-independent apoptosis. Taken together, our results show that Aloe vera extract induce HepG2 apoptosis by ATP depletion-related impairment of mitochondria, which is caspase-independent.

• Advances in Traditional Medicine
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• 제4권3호
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• pp.171-178
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• 2004

Ginseng radix, the root of Panax ginseng C. A. Meyer (Araliaceae), is a medicinal plant used world-widely and has been reported to have various biological effects. To investigate the effects of Ginseng radix on HL-60 cell apoptosis, MTT assay, DNA fragmentation assay and flow cytometry were performed on HL-60 cells. Cells were treated with Ginseng radix at different concentrations $(10^{-4},\;10^{-3}\;and\;10^{-2};\;dilution\;rate)$. Ginseng radix significantly induced cells apoptosis with a time- and dose-dependent manner. To determine whether Ginseng radix-induced apoptosis is due to increase of tumor necrosis factor $(TNF-{\alpha})$ secretion, enzyme-linked immunosorbent assay was performed on HL-60 cells. Unexpectedly, Ginseng radix $(96\;{\pm}\;5\;pg/ml)$ significantly decreased the $TNF-{\alpha}$ secretion compared with control $(174\;{\pm}\;14\;pg/ml)$. Furthermore, Ginseng radix with $rIFN-{\gamma}$ synergistically increased nitric oxide production in mouse peritoneal macrophages. Taken together, our data indicate that Ginseng radix induce apoptosis on HL-60 cells without increase of $TNF-{\alpha}$ secretion and could be used for a supplementary remedy of cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7069-7074
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• 2013

Programmed cell death is a basic cellular process that is critical to maintaining tissue homeostasis. In contrast to apoptosis, necrosis was previously regarded as an unregulated and uncontrollable process. However, as research has progressed, necrosis, also known as necroptosis or programmed necrosis, is drawing increasing attention, not least becasu of its possible impications for cancer research. Necroptosis exhibits a unique signaling pathway that requires the involvement of receptor interaction protein kinases 1 and 3 (RIP1 and RIP3), mixed lineage kinase domain-like (MLKL), and phosphoglycerate mutase 5 (PGAM5) and can be specifically inhibited by necrostatins. Not only does necroptosis serve as a backup cell death program when apoptosis is inhibited, but it is now recognized to play a pivotal role in regulating various physiological processes and the pathogenesis of a variety of human diseases such as ischemic brain injury, immune system disorders and cancer. The control of necroptosis by various defined trigger factors and signaling pathways now offers the opportunity to target this cellular process for therapeutic purposes. The purpose of this paper is to review current findings concerning the connections between various trigger factors and the RIP1/RIP3 signaling pathway as it relates to necroptosis.

• 한국미생물·생명공학회지
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• 제50권3호
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• pp.387-394
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• 2022

Verotoxin-1 (VT-1) or Shiga-like toxin 1 (Stx-1) is produced by enterohemorrhagic Escherichia coli (EHEC) and is an AB5 holotoxin with a strong inhibitor of protein synthesis. VT-1 is a type 2 ribosome-inactivating protein (RIP) that has been shown to have cytotoxic and anticancer potential by inducing necrosis, apoptosis, and cell cycle arrest, making it a promising antitumor candidate. Here, we tested the cytotoxicity of VT-1 on CaCo2 and NCM425 cell lines and the results showed that VT-1 was more potent on CaCo2. Morphological changes were also evaluated on the cellular level and the results showed that VT-1 caused a decrease in viable cell count, altered cell membrane permeability, and an increase in total nuclear intensity. On the other hand, VT-1 displayed a lesser impact on mitochondrial membrane potential (MMP) and cytochrome c release. On the expression of caspases 3 and 9, VT-1 exhibited an insignificant effect on both which alongside the mitochondrial membrane potential (MMP) and cytochrome c results, might indicate that CaCo2 suffered from the necrosis process as a mechanism of cell death after exposure to VT-1.

• Journal of Pharmaceutical Investigation
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• 제33권4호
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• pp.261-265
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• 2003

Tumor necrosis facto-related apoptosis-inducing ligand (TRAIL) is a recently identified member of the tumor necrosis factor cytokine superfamily. TRAIL has been shown to induce apoptosis in a number of tumor cells whereas cells from most of normal tissues are highly resistant to TRAIL-induced apoptosis. These observations have raised considerable interest in the use of TRAIL in tumor therapy. In this study we report the biodistribution fates and serum expression pattern of plasmid DNA encoding TRAIL (pTRAIL) delivered in erythrocyte ghosts (EG). pTRAIL was loaded into EG by electroportion in a hypotonic medium The mRNA expression of pTRAIL was prolonged following delivery in EG-encapsulated forms. EG containing pTRAIL showed significant levels of mRNA expression in the blood over 9 days. The organ expression patterns of pTRAIL delivered via EG, however, did not significantly differ from those of naked pTRAIL, indicating that the expression-enhancing effect of EG containing pTRAIL was localized to the blood. These results suggest that pTRAIL-loaded EG might be of potential use in the treatment of hematological diseases such as TRAIL-sensitive leukemia.

• 대한수의학회지
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• 제44권3호
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• pp.329-334
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• 2004

Ischemia/reperfusion(I/R) injury of the rat testis causes germ cell death and infiltration of inflammatory cells. To investigate the mechanism of germ cell death in torsion of the rat testis, apoptosis and macrophage activation were studied using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL) method and immunohistochemistry in the testes of Sprague-Dawley rats subjected to 1.5 h of ischemia, followed by 0, 1, 3, 6, 12, 24, 48 and 96 h of reperfusion. Apoptotic, TUNEL-positive cells were found at the base of the seminiferous epithelia after I/R. TUNEL-positive cells were significantly increased 6 h after repair of the torsion, and there was a significant peak in apoptosis 24 h after reperfusion, as compared with normal or sham-operated controls. In contrast, histological evidence of germ cell necrosis in the seminiferous tubules was first visible 24 h after reperfusion. In the testis of sham-operated rats, ED2-positive resident macrophages were found diffusely in the interstitial space, while ED1-positive monocyte-like macrophages were rarely found. After I/R, ED1-positive cells were significantly increased beginning 12 h after reperfusion, while ED2-positive immunoreactivity did not change during the experimental period. Together, the results of this study confirmed that increased numbers of ED1-positive macrophages, but not resident ED2-positive macrophages, infiltrated the interstitial space surrounding damaged tubules and induced germcell death.

• 생명과학회지
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• 제19권12호
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• pp.1705-1711
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• 2009

항암제 내성을 획득한 암세포는 많은 경우 다양한 세포 독성 물질에 대해 교차 내성을 나타낸다. 그러나 온열 치료가 내성 획득 종양에 적용 될 때의 종양 세포의 사멸 효과는 알려져 있지 않다. 본 연구는 시스플라틴에 내성을 갖는 위암 세포 주, SNU601/Cis2이 열충격에 반응하는 민감도와 세포 사멸 방식을 조사함으로써 약물 내성 종양의 온열 치료 효과를 예측하고자 하였다. 정상 위암 세포 주 SNU601/WT은 열충격에 매우 민감하게 반응하며 apoptosis로 사멸하지만, 내성 위암 세포 주 SNU601/Cis2는 미열충격에 내성을 나타내었으며 고열충격에 노출되자 necrosis로 사멸하였다. 또한 SNU601/Cis2에서 necrosis의 발생은 열충격에 의한 JNK1/2의 활성화와 HSP27의 발현저하 현상과 관련되어 있었다. Necrosis의 유도는 세포막 파괴에 의해 세포 내부 물질의 방출로 인한 주변 조직의 염증반응을 수반하는데, 이러한 염증 반응은 암의 성장을 촉진하고 암의 성상을 심화시키는 것으로 보고되고 있다. 이러한 관점에서, 온열 치료가 약물 치료와 병행 될 경우에는 교차 내성과 necrosis로 인한 역효과를 방지하기 위하여, 그 적용이 주의 깊게 이루어져야 할 것으로 판단된다.