• 생명과학회지
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• 제20권4호
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• pp.572-577
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• 2010

본 연구에서 60 주령된 쥐를 대상으로 8 주간 트레드밀 운동강도와 근섬유 유형에 따른 apoptosis 관련 인자를 연구하였다. 결론적으로 노후된 쥐의 apoptosis 관련 인자는 운동강도와 근육유형에 따라 차이가 났다. EDL이 Soleus보다 apoptosis인자의 변화가 운동강도에 의해 더 민감하게 나타났다. 또한 casepase-3의 발현은 고강도 운동에서 나타났으나 DNA 절편화는 나타나지 않았기에 실질적인 apoptosis는 발생되지 않았다. 이는 노후된 개체의 운동 수행방법으로 고강도 운동은 적합하지 않으며, 고강도 운동상태를 지속할 경우 근섬유의 apoptosis를 유도할 가능성이 있다는 것을 시사한다. 그러므로 추후 실험에서는 운동시간과 빈도에 의한 apoptosis의 time-course impact 관찰과 외재적 경로(caspase independent apoptosis)에서의 apoptosis 관련 단백질 변화에 대한 연구를 통하여 노화에 의한 근손실의 원인 규명이 요구된다.

• 한국식품영양과학회지
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• 제37권9호
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• pp.1114-1119
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• 2008

ECCG는 녹차 카테킨의 주요 성분으로 항산화작용으로 인한 항암작용이 보고되고 있다. 본 연구는 EGCG가 전이성이 강한 인체 유방암 세포인 MDA-MB-231의 세포사멸에도 영향을 주는지를 알아보고자 하였다. 인체 유방암 세포 배양액에 EGCG를 0, 5, 10, $20\;{\mu}M$로 첨가시켜, 세포사멸과 관련된 단백질들의 단백질과 mRNA 발현, caspase-3 활성을 관찰하였다. EGCG 첨가 농도가 $5\;{\mu}M$ 이상부터 세포사멸을 억제하는 단백질인 bcl-2의 단백질과 mRNA 발현이 감소하였으며, 세포사멸을 유도하는 단백질인 bax의 단백질과 mRNA 발현은 유의적으로 증가하여 결과적으로 EGCG 첨가에 따라 bcl-2/bax의 비율이 유의적으로 감소하였다. 또한 세포사멸의 마지막 단계인 caspase-3의 활성은 EGCG 농도가 증가할수록 유의적으로 증가하였다. 본 연구 결과를 종합해 보면 전이성이 강한 인체 유방암 세포 MDA-MB-231에서 EGCG는 암세포에서 bcl-2의 발현은 억제시키고 bax의 발현은 증가시키며, caspase-3의 활성을 증가시켜 세포사멸을 유도하는 것으로 확인하였다.

• Molecules and Cells
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• 제24권3호
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• pp.378-387
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• 2007

The Bcl-2 family of proteins interacts at the mitochondria to regulate apoptosis. However, the anti-apoptotic Bcl-2 and $Bcl-X_L$ are not completely localized to the mitochondria. In an attempt to generate Bcl-2 and $Bcl-X_L$ chimeras that are constitutively localized to the mitochondria, we substituted their C-terminal transmembrane tail or both the C-terminal transmembrane tail and the adjacent loop with the equivalent regions from Bak or Bax mutant (BaxS184V) as these regions determine the mitochondrial localization of Bak and Bax. The effects of these substitutions on subcellular localization and their activities were assessed following expression in HeLa and CHO K1 cells. The substitution of the C-terminal tail or the C-terminal tail and the adjacent loop of Bcl-2 with the equivalent regions from Bak or the Bax mutant resulted in its association with the mitochondria. This change in subcellular localization of Bcl-2 chimeras triggered cells to undergo apoptotic-like cell death. The localization of this Bcl-2 chimera to the mitochondria may be associated with the disruption of mitochondrial membrane potential. Unlike Bcl-2, the loop structure adjacent to the C-terminal tail in $Bcl-X_L$ is crucial for its localization. To localize the $Bcl-X_L$ chimeras to the mitochondria, the loop structure next to the C-terminal tail in $Bcl-X_L$ protein must remain intact and cannot be substituted by the loop from Bax or Bak. The chimeric $Bcl-X_L$ with both its C-terminal tail and the loop structure replaced by the equivalent regions of Bak or Bax mutant localized throughout the entire cytosol. The $Bcl-X_L$ chimeras that are targeted to the mitochondria and the wild type $Bcl-X_L$ provided same protection against cell death under several death inducing conditions.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7065-7068
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• 2014

To investigate the anti-proliferative mechanism of mangiferin in a human nasopharyngeal carcinoma cell line, CNE2 cells were incubated with different concentrations of mangiferin (12.5, 25, 50, 100, 150 and $200{\mu}M$) or with PBS as a control for 72 hours. Analyses were made of the cell cycle and apoptosis with measurement of mRNA and protein levels of two apoptosis-related genes, Bcl-2 and Bax. Flow cytometry assays showed mangiferin could inhibit CNE2 cell proliferation via G2/M arrest and induction of early apoptosis. Real time PCR and Western blotting showed the mRNA and protein level of Bcl-2 to be down-regulated, while those of Bax were upregulated, when CNE2 cells were treated with mangiferin. This investigation indicated anti-proliferation effects of mangiferin through induction of cell apoptosis regulated by Bcl-2 and Bax expression.

• 한방재활의학과학회지
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• 제19권3호
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• pp.1-13
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• 2009

Objectives : This study evaluates neuroprotective effect of Gastrodiae Rhizoma on apoptosis in the cerebral infarct. Methods : Cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method in Sprague-Dawley rats. Then ethanol extract of Gastrodiae Rhizoma was administered orally for 3 days. Infarct area and volume were evaluated with TTC staining. Neuronal apoptosis was identified with TUNEL labeling. Apoptosis modulatory effect was observed with immunohistochemical Bax, Bcl-2, iNOS, and MMP-9 expressions in penumbra. Results : 1. Ethanol extract of Gastrodiae Rhizoma reduced infarct size partly and volume significantly of in the MCAO rat brain. 2. Ethanol extract of Gastrodiae Rhizoma reduced TUNEL positive cell ratio in the penumbra of MCAO rat brain significantly. 3. Ethanol extract of Gastrodiae Rhizoma suppressed Bax, iNOS and MMP-9 expression in the penumbra of MCAO rat brain significantly. 4. Ethanol extract of Gastrodiae Rhizoma did not change Bcl-2 expression in the penumbra of MCAO rat brain. But expression ratio of Bcl-2 against Bax was increased in the Gastrodiae Rhizoma group. Conclusions : These results suggest that Gastrodiae Rhizoma plays an anti-apoptotic neuroprotective effect through suppression of Bax, iNOS, and MMP-9 expressions and relative up-regulation of Bcl-2 in the ischemic brain tissue.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.3195-3199
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• 2014

Inonotus obliquus is a medicinal mushroom that has been used as an effective agent to treat various diseases such as diabetes, tuberculosis and cancer. Inotodiol, an included triterpenoid shows significant anti-tumor effect. However, the mechanisms have not been well documented. In this study, we aimed to explore the effect of inotodiol on proliferation and apoptosis in human cervical cancer HeLa cells and investigated the underlying molecular mechanisms. HeLa cells were treated with different concentrations of inotodiol. The MTT assay was used to evaluate cell proliferating ability, flow cytometry (FCM) was employed for cell cycle analysis and cell apoptosis, while expression of cyclinE, p27, bcl-2 and bax was detected by immunocytochemistry. Proliferation of HeLa cells was inhibited by inotodiolin a dose-dependent manner at 24h (r=0.9999, p<0.01). A sub-$G_1$ peak (apoptotic cells) of HeLa cells was detected after treatment and the apoptosis rate with the concentration and longer incubation time (r=1.0, p<0.01), while the percentage of cells in S phase and $G_2$/M phase decreased significantly. Immunocytochemistry assay showed that the expression of cyclin E and bcl-2 in the treated cells significantly decreased, while the expression of p27 and bax obviously increased, compared with the control group (p<0.05). The results of our research indicate that inotodiol isolated from Inonotus obliquus inhibited the proliferation of HeLa cells and induced apoptosis in vitro. The mechanisms may be related to promoting apoptosis through increasing the expression of bax and cutting bcl-2 and affecting the cell cycle by down-regulation the expression of cyclin E and up-regulation of p27. The results further indicate the potential value of inotodiol for treatment of human cervical cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.4893-4898
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• 2016

Objective: To investigate the impact of a Croton tiglium extract on cellular proliferation and apoptosis in a non-small cell lung cancer cell line (A549) in vitro. Methods: A Croton tiglium seed methanol extract was prepare and assessed for effects on A549 cells regarding cellular proliferation, apoptotic rates, and expression of apoptosis related genes and proteins using real-time PCR and immunofluorescence. Results: The tested Croton tiglium extract inhibited A549 cell proliferation in a dose- and time-dependent manner, with significant elevation of apoptotic indexes at various concentrations after 24 h. In addition, rates in both early and late stages were higher in treated than untreated groups, the $100{\mu}g/ml$ dose causing the highest levels of apoptosis. RT-PCR showed that A549 cells treated with $100{\mu}g/ml$ Croton tiglium extract for 24 h has markedly higher Bax mRNA expression levels and obviously lower Bcl-2 expression levels than controls, equivalent results being observed for proteins by immunofluorescence. However, the mRNA expression levels of Fas and caspase-8 were not significantly altered. Conclusion: A Croton tiglium extract can inhibit proliferation of A549 cells and promote apoptosis though Bax/Bcl-2 pathways.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.5031-5035
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• 2016

Docetaxel, recognized as a stabilizing microtubule agent, is frequently administrated as a first line treatment for prostate cancers. Due to high side effects of monotherapy, however, combinations with novel adjuvants have emerged as an alternative strategy in cancer therapy protocols. Here, we investigated the combined effects of stattic and docetaxel on the DU145 prostate cancer cell line. Cytotoxicity was evaluated by MTT assay. To understand molecular mechanisms of stattic action, apoptotic related genes including Bcl-2, Mcl-1, Survivin and Bax were evaluated by real-time RT-PCR. Alteration in the expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 genes and Bax/Bcl-2 ratio were investigated via the $2^{{\Delta}{\Delta}CT}$ method. The $IC_{50}$ values for docetaxel and stattic were $3.7{\pm}0.9nM$ and $4.6{\pm}0.8{\mu}M$, respectively. Evaluation of key gene expression levels revealed a noticeable decrease in antiapoptotic Bcl-2 and Mcl-1 along with an increase in pro-apoptotic Bax mRNA levels (p<0.05). Our results suggest that combination of a STAT3 inhibitor with doctaxel can be considered as a potent strategy for induction of apoptosis via increasing Bax mRNA expression.

• Asian Pacific Journal of Cancer Prevention
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• 제13권8호
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• pp.3857-3862
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• 2012

A total of 96 cases of invasive breast ductal carcinoma were examined for immunohistochemical expression of Bax and Bcl-2 in the epithelial tumor cells and endothelial cells of the blood vessels. We also investigated the association between both proteins in the epithelium in relation to tumor characteristics such as tumor size, grade, lymph node involvement, microvessel density (MVD), hormonal receptors expression and c-erbB-2 overexpression. Bax expression showed a significant association between tumor and endothelial cells (p<0.001) while Bcl-2 expression in tumor cells was inversely associated with that in the endothelial cells (p<0.001). Expression of Bcl-2 in tumor cells was strongly associated with expression of estrogen and progesterone receptors (p=0.003 and p=0.004, respectively). In addition, intratumoral MVD was significantly higher than peritumoral MVD (p<0.001) but not associated with Bax or Bcl-2 expression and other tumor characteristics. We concluded that the number of endothelial cells undergoing apoptosis was in direct linkage with the number of apoptotic tumor cells. Anti-apoptotic activity of the surviving tumor cells appears to propagate cancer progression and this was influenced by the hormonal status of the cells. Tumor angiogenesis was especially promoted in the intratumoral region and angiogenesis was independent of anti-apoptotic activity.

• 한방안이비인후피부과학회지
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• 제29권3호
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• pp.159-167
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• 2016

Prostate cancer is one of the most common non-skin cancers in men. Armeniacae Amarum Semen has traditionally been used for the treatment of inflammation diseases, leprosy, leucoderma, and tumors. Apoptosis, which is also known as programmed cell death, is an important mechanism in cancer treatment.Objectives : We observed whether an aqueous extract of Armeniacae Amarum Semen induces apoptotic cell death in human DU145 prostate cancer cells.Methods : We treated DU145 cells with Armeniacae Amarum Semen extract and investigated characteristics of apoptosis. And investigated whether treated with Armeniacae Amarum Semen extract increased Bax mRNA expression, Bcl-2 mRNA expression, caspase-3 enzyme activity and their protein level.Results : We have shown that Armeniacae Amarum Semen extract can induce apoptotic cell death in human DU145 prostate cancer cells by caspase-3 activation through the down-regulation on Bcl-2 expression and the up-regulation on Bax expression.Conclusions : It can be expected that an aqueous extract of Armeniacae Amarum Semen may offer a valuable means for the treatment of prostate cancers.