• The Korean Journal of Physiology and Pharmacology
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• 제14권6호
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• pp.407-412
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• 2010

3-Deazaadenosine (DZA), a potent inhibitor of S-adenosylhomocysteine hydrolase, was previously proposed to induce intrinsic apoptosis in human leukemic cells. In the present study, we analyzed the mechanism underlying the DZA-induced intrinsic apoptotic pathway. DZA activated typical caspase-dependent apoptosis in HL-60 cells, as demonstrated by an accumulation of hypo-diploidic cells, the processing of multiple procaspases and an inhibitory effect of z-VAD-Fmk on this cell death. During DZA-induced apoptosis, cytochrome c (cyt c) was released into the cytosol. This was neither prevented by z-VAD-Fmk and nor was it associated with the dissipation of mitochondrial membrane potential (${\Delta}{\Psi}_m$). Prior to the release of cyt c, BAX was translocated from the cytosol to mitochondria and underwent oligomerization. Finally, the overexpression of BCL-XL protected HL-60 cells from apoptosis by blocking both the cyt c release and BAX oligomerization. Collectively, these findings suggest that DZA may activate intrinsic apoptosis by stimulating BAX activation and thereby the release of cyt c.

• Clinical and Experimental Reproductive Medicine
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• 제29권3호
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• pp.195-200
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• 2002

Objective : To investigate the expression of apoptosis related proteins and apoptotic cells on the human ovarian follicles. Materials and Methods: Thirty five Formalin-fixed paraffin-embedded human ovarian tissue blocks were selected from the surgical pathology files of the department of pathology, College of Medicine, Yonsei University, for the period from 1996 to 1998. All specimen were from premenopausal women aged from $32{\sim}45$. Ovarian tissues were collected from the patients performing hysterectomy for benign uterine diseases. Immunohistochemical staining was performed for the detection of DNA fragmented cell, Bcl-2, Bax, Fas and Fas-ligand. Results: Bcl-2 and bax were not expressed on the surrounding cells and oocyte of the primary, primordial and preantral follicles. Fas and Fas-ligand (Fas-L) were not expressed on the surrounding cells on the primordial and primary follicles. But expressed on the surrounding granulosa cells and oocyte in the primordial and primary follicles. In the healthy follicles, Bcl-2 was expressed on the granulosa cells, however, Bax was not expressed. DNA fragmented cells were expressed on the inner granulosa cell layer of atretic follicles. Conclusion: Fas, Fas-ligand, and Bax may be responsible for the follicular atresia and Bcl-2 may be involved in the follicular survival in the human ovary.

• 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.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.734-740
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• 2005

The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8617-8622
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• 2014

The goal of this study was to establish paclitaxel resistant MCF-7 cells, as in vitro model, to identify the molecular mechanisms leading to acquired chemoresistance in breast cancer cells. Resistant cells were developed by stepwise increasing exposure to paclitaxel. Gene expression levels of Bax and Bcl-2 along with protein levels of caspase-8 and caspase-9 were evaluated in two resistant cell lines (MCF-7/Pac64 and MCF-7/Pac5 nM). Morphological modifications in paclitaxel resistance cells were examined by light microscopy and fluorescence activated cell sorting (FACS). As an important indicator of resistance to chemotheraputic agents, the Bcl-2/Bax ratio showed a significant increase in both MCF-7/Pac5nM and MCF-7/Pac 64nM cells (p<0.001), while caspase-9 levels were decreased (p<0.001) and caspase-8 was increased (p<0.001). FACS analysis demonstrated that MCF-7/Pac64 cells were smaller than MCF-7 cells with no difference in their granularity. Our results support the idea that paclitaxel induces apoptosis in a mitochondrial-dependent manner. Identifying breast cancer patients with a higher Bcl-2/Bax ratio and caspase 9 level and then inhibiting the activity of these proteins may improve the efficacy of chemotheraputic agents.

• 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.

• 대한한방부인과학회지
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• 제18권1호
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• pp.29-44
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• 2005

To address the ability of Herba Portulacae(HP) to induce cell death, we investigated the effect of HP on cell viability. Twenty-four hours later, loss of viability occurred following HP exposure in a dose-dependent manner. The treatment of HP, a commonly used herb formulation in Korea, Japan and China, caused a decrease in cell viability. HP also resulted in apoptotic morphology a brightly blue-fluorescent condensed nuclei by Hoechst 33258-staining, and reduction of cell volume. Our results show that 2mg/ml HP induces mitochondria membrane potential collapse. Immunoblotting data also shows that the expression of Bcl-2, antiaoptotic protein, decrease by the addition of HP. This GFP-Bax overexpression system shows that an important pro-apoptotic Bcl-2-family protein, Bax is translocated to mitochondria by the addition of 2mg/ml HP. Inerestingly, MAPK inhibitor study shows that p38 MAPK inhibitor, SB203580 inhibits HP-induced cell death and caspase-3 activation in HP-treated HeLa cells. Furthermore, HP transiently but significantly induces p38 activation. But P38 MAPK inhibitor does not have any effect on the translocation of Bax. Considering these results, HP induces apoptosis via p38 MAPK activation. But the pathway does not involve the translocation of Bax.

• Clinical and Experimental Pediatrics
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• 제62권3호
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• pp.95-101
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• 2019

Purpose: Increased apoptosis was recently found in the hypertrophied left ventricle of spontaneously hypertensive rats (SHRs). Although the available evidence suggests that apoptosis can be induced in cardiac cells by various insults including pressure overload, cardiac apoptosis appears to result from an exaggerated local production of angiotensin in adult SHRs. Altered expressions of Bcl associated X (Bax), Bcl-2, chemokine receptor (CCR)-2, monocyte chemoattractant protein (MCP)-1, transforming growth factor $(TGF)-{\beta}1$, phosphorylated extracellular signal-regulated kinases (PERK), and connexin 43 proteins, and kallikrein mRNA were investigated to explore the effects of losartan on the SHR model. Methods: Twelve-week-old male rats were grouped as follows: control (C), SHR (hypertension: H), and losartan (L; SHRs were treated with losartan [10 mg/kg/day] for 5 weeks). Western blot and reverse transcription polymerase chain reaction assays were performed. Results: Expression of Bax, CCR-2, MCP-1, $TGF-{\beta}1$, PERK, and connexin 43 proteins, and kallikrein mRNA was significantly increased in the H group compared to that in the C group at weeks 3 and 5. Expression of Bax, CCR-2, MCP-1, $TGF-{\beta}1$, and connexin 43 proteins and kallikrein mRNA was significantly decreased after losartan treatment at week 5. PERK protein expression was significantly decreased after losartan treatment at weeks 3 and 5. Bcl-2 protein expression was significantly decreased in the H group compared to that in the C group at weeks 3 and 5. Conclusion: Losartan treatment reduced expression of Bax, CCR-2, MCP-1, $TGF-{\beta}1$, PERK, and connexin 43 proteins, and kallikrein mRNA in SHRs, along with decreased inflammation and apoptosis.

• 운동영양학회지
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• 제15권4호
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• pp.173-182
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• 2011

Post-traumatic stress disorder (PTSD) is a stress-related mental disorder induced by severe external stressors such as assault, disaster or severe accident. We investigated the effects of treadmill exercise on short-term memory in relation to apoptosis and cell proliferation in the hippocampus following PTSD. Stress to the pregnant rats was induced by exposure of maternal rats to the hunting dog in an enclosed room. Exposure time was 10 min, repeated three times per day, with 1 hour interval. Exposure of maternal rats to the hunting dog was continued 7 days after pregnancy until delivery. The pregnant rats in the exercise groups were forced to run on a treadmill for 30 min once a day for the same duration of stress exposure. Step-down avoidance task for short-term memory, western blot for Bcl-2, Bax, and immunohistochemistry for caspase-3, 5-bromo-2'-deoxyuridine (BrdU), and Ki-67 were conducted. Maternal rats exposed to stress during pregnancy showed short-term memory impairment. Expressions of Bax, Bcl-2, ratio of Bax to Bcl-2, and caspase-3 in the hippocampus were increased in the PTSD rats. Cell proliferation in the hippocampal dentate gyrus was decreased in the PTSD rats. Treadmill exercise alleviated short-term memory impairment and suppressed expressions of Bax, the ratio of Bax to Bcl-2, and caspase-3. Treadmill exercise also increased cell proliferation. The present results indicate that treadmill exercise alleviated PTSD-induced short-term memory impairment by suppressing apoptotic cell death and enhancing cell proliferation in the hippocampus.