• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7561-7568
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• 2013

Galectin-3 (Gal-3) is a carbohydrate-binding protein which is thought to be involved in cancer progression but its contribution to epithelial ovarian cancer (EOC) remains unclear. The present study sought to determine the role of Gal-3 in chemoresistance of the human SKOV-3 ovarian cancer cell line to paclitaxel (PTX) using recombinant human Gal-3 (rhGal-3) and PectaSol-C modified citrus pectin (Pect-MCP) as a specific Gal-3 competitive inhibitor. Our results showed 41% increased cell proliferation, 36% decreased caspase-3 activity and 33.6% increased substrate-dependent adhesion in the presence of rhGal-3 compared to the control case (p<0.001). Treatment of cells with a non-effective dose of PTX (100nM) and 0.1% Pect-MCP in combination revealed synergistic cytotoxic effects with 75% reduced cell viability and subsequent 3.9-fold increase in caspase-3 activity. Moreover, there was 39% decrease in substrate-dependent adhesion compared to control (p<0.001). These results suggest that inhibition of Gal-3 could be a useful therapeutic tool for combination therapy of ovarian cancer.

• 환경생물
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• 제26권3호
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• pp.214-219
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• 2008

Changes in stress-associated biomolecules can be used as an important criterion for assessing the levels of environmental pollution because living organisms demonstrate contamination-stimulated stress responses. This study was conducted to determine the environmental status of Masan-Jinhae Bay, Korea, and its effects on marine organisms by investigating the endoplasmic reticulum (ER) dysfunction in the organs of the flat fish, Limanda yokohamae. ER dysfunction was evaluated via Western blot analysis of the ER stress proteins, immunoglobulin heavy chain binding protein (BiP) and C/EBP-homologous protein (CHOP), and the ER stress-associated protein caspase-12. The results showed that the amount of BiP and CHOP immunoreactivity in the flat fish from the bay area was much greater than that from the Gangneung, as a reference site. Similar to the ER stress proteins, the immunoreactivity of caspase-12 was also found to be elevated in the bay area when compared with that of Gangneung. These data suggest that the environmental status of Masan-Jinhae Bay induces the ER stress response, which is able to lead to phenotypic changes in marine organisms including fish.

• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5189-5193
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• 2016

Objective: Dorema glabrum Fisch. & C.A. Mey is a perennial plant that has several curative properties. Anti-proliferative activity of seeds of this plant has been demonstrated in a mouse fibrosarcoma cell line. The aim of the present study was to evaluate cytotoxicity of D. glabrum root extracts in a human gastric adenocarcinoma (AGS) cell line and explore mechanisms of apoptosis induction, cell cycle arrest and altered gene expression in cancer cells. Materials and Methods: The MTT assay was used to evaluate IC50 values, EB/AO staining to analyze the mode of cell death, and flow cytometry to assess the cell cycle. Quantitative real-time polymerase chain reaction (qRT-PCR) amplification was performed with apoptosis and cell cycle-related gene primers, for cyclin D1, c-myc, survivin, VEGF, Bcl-2, Bax, and caspase-3 to determine alteration of gene expression. Results: Our results showed that n-hexane and chloroform extracts had greatest toxic effects on gastric cancer cells with IC50 values of $6.4{\mu}g/ml$ and $4.6{\mu}g/ml$, respectively, after 72 h. Cell cycle analysis revealed that the population of treated cells in the G1 phase was increased in comparison to controls. Cellular morphological changes indicated induction of apoptosis. In addition, mRNA expression levels of Bax and caspase-3 were increased, and of bcl-2 survivin, VEGF, c-myc and cyclin D1 were decreased. Conclusion: Our study results suggest that D. glabrum has cytotoxic effects on AGS cells, characterized by enhanced apoptosis, reduced cell viability and arrest of cell cycling.

• 동의생리병리학회지
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• 제23권6호
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• pp.1368-1378
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• 2009

Unfolded protein response (UPR) is an important genomic response to endoplasmic reticulum (ER) stress. The ER response is characterized by changes in specific proteins, induction of ER chaperones and degradation of misfolded proteins. Also, the pathogenesis of several diseases like Alzheimer's disease, neuronal degenerative diseases, and diabetes reveal the role of ER stress as one of the causative mechanisms. Borneolum has been used for neuronal disease in oriental medicine. In the present study, the protective effect of borneolum on thapsigargin-induced apoptosis in rat C6 glial cells. Treatment with C6 glial cells with 5 uM thapsigargin caused the loss of cell viability, and morphological change, which was associated with the elevation of intracellular $Ca^{++}$ level, the increase in Grp78 and CHOP and cleavage of pro-caspase 12 Furthermore, thapsigargin induced Grp98, XBP1, and ATF4 protein expression in C6 glial cells. Borneolum reduced thapsigargin-induced apoptosis through ER pathways. In the ER pathway, borneolum attenuated thapsigargin-induced elevations in Grp78, CHOP, ATF4, and XBP1 as well as reductions in pro-caspase 12 levels. Also, our data showed that borneolum protected thapsigargin-induced cytotoxicity in astrocytes from rat (P3) brain. Taken together, our data suggest that borneolum is neuroprotective against thapsigargin-induced ER stress in C6 glial cells and astrocytes. Accordingly, borneolum may be therapeutically useful for the treatment of thapsigargin-induced apoptosis in central nervous system.

• Applied Biological Chemistry
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• 제50권1호
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• pp.63-67
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• 2007

본 연구에서는 석곡(Dendrobium nobile Lindl.) 추출물에 대한 신경세포 보호 효과를 확인하기 위하여 $H_{2}O_{2}$에 의해 유도된 스트레스 상태의 PC12 세포주에서 MTT-dye reduction assay와 LDH release assay를 이용하였다. MTT reduction assay 결과, 스트레스 상태에서는 47%의 생존율을 보인데 반하여 석곡 추출물 50${\mu}$g/ml 농도로 처리하였을 때 99.5%의 높은 세포 생존율을 확인할 수 있었다. 이 결과는 LDH release assay 에서도 일치하는 결과를 확인하였다. 그리고 광학 현미경을 이용한 형태학적 변화를 관찰한 결과에서도 신경돌기의 출현 유도를 통한 신경세포 생존을 확인 할 수 있었다. 또한 핵의 변화에 미치는 영향을 관찰한 결과 정상 세포의 핵은 타원형의 온전한 핵 모양을 나타낸 반면 산화적 손상을 입은 세포는 apoptotic body가 핵 주변에 나타나는 전형적인 apoptosis를 나타내었고 석곡 추출물을 처리한 결과 핵의 condersation현상과 fragmentation이 현저히 감소함을 확인 할 수 있었다. 그리고 $H_{2}O_{2}$에 의한 capase-3의 활성은 석곡 추출물을 50${\mu}$g/ml 처리 후 약 1.1배 이하로 감소함을 보였다.

• 한국식품영양과학회지
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• 제40권12호
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• pp.1654-1661
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• 2011

AKE의 농도별 처리가 인체 유방암 세포 MDA-MB-231의 세포사멸에 미치는 영향을 확인하기 위하여 세포 화학적인 방법인 MTT 분석, 이중 핵 염색법(Hoechst 33342/EtBr staining), FACS를 통하여 세포사멸을 관찰하였다. MTT 분석 결과, 150 ${\mu}g$/mL 처리 군에서 대조군에 대비하여 약 50%의 세포사멸을 나타내었으며 세포사멸이 농도 의존적으로 증가되었고(p<0.05), 이중 핵 염색법을 이용하여 세포사의 구분 결과 능동적 세포예정사인 apoptosis가 농도 의존적으로 급격히 증가하였으며(p<0.05), 특히 150 ${\mu}g$/mL 처리군에서 현저한 증가율을 나타내었다. 보다 더 명확한 세포사멸을 확인하기 위하여 FACS를 이용한 apoptosis 측정 결과, 처리군 간 크게 차이를 보이며 농도 의존적으로 증가되었다. 세포사멸관련 mRNA 유전자 발현을 관찰한 결과, 세포사멸 억제 유전자 Bcl-2는 처리농도가 증가할수록 유의적 증가를 보였으며(p<0.05), 세포사멸 유도 유전자 Bax는 유의적 감소를 나타내었다(p<0.05). 세포사멸의 지표인 Bcl-2/Bax의 비율은 농도 의존적인 감소를 나타내었으며(p<0.05), 세포사멸유도의 마지막 단계의 실행자인 caspase-3의 활성도 첨가 농도 의존적으로 증가하여 세포사멸을 유도하는 것으로 확인되었다(p<0.05). 결론적으로, AKE는 유방암 세포 MDA-MB-231의 세포사멸을 유도하는 것으로 나타나 신선초의 항암효과의 가능성을 제시해주었다. 향후 in vivo 실험에서도 신선초의 항암효과에 대한 심층적 연구가 이뤄져야 할 것으로 사료된다.

• 생명과학회지
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• 제12권4호
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• pp.452-459
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• 2002

호중의 세포사멸은 자연적으로 일어나지만 여러 자극에 의한 신호에 의하여 증가하거나 지연된다. 본 연구에서는 세포 내 단백질 분비과정을 억제한다고 알려진 BFA가 호중구의 자연 세포사멸 및 세포사멸 지연에 어떠한 기작으로 작용하는가를 연구하였다. 호중구의 세포사멸은 사람 말초 혈액으로부터 분리하여 세포 배양 20시간 후 형태 변화, annexin V and propidium iodide의 염색, 및 DNA 전기영동 등으로 조사하였다. BFA는 농도 의존형으로 호중구의 세포사멸을 증가시킨다. CM-CSF나 LPS에 의한 세포사멸의 지연도 BFA에 의하여 억제되었다. 그러나 BFA의 영향은 db-cAMP, dexamethasone, 및 IL-8을 처리한 세포에서는 큰 영향을 받지 않았다. PKC-5의 억제제인 rottlerin에 의한 세포사멸의 지연은 BFA에 의하여 감소하였다. 그러나 BFA에 의한 세포사멸의 유도는 caspase-3 억제제인 zDEVD-fmk에 의하여는 영향을 받지 않았다. 한편, 세포사멸 억제에 관여하는 Mcl-1 단백질의 발현은 BFA의 처리에 의하여 감소하였다. 이들 결과들은 세포 내 단백질 분비 과정의 억제가 호중구의 세포사멸에 관여하며 이들의 작용은 Mcl-1 발현의 조절에 의한다는 것을 제시하고 있다.

• 동의신경정신과학회지
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• 제20권2호
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• pp.111-120
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• 2009

Objectives : This study was performed to investigate the effects of Needle Electrode Electrical Stimulation (NEES) on ischemia-induced cerebrovascular accidents. After obstruction and reperfusion of ${\ast}{\ast}$ arteries in white mice, the amounts of necrosis and inflammation related substances IL-6, Caspase-3, and PARP, C-fos were measured in neurons of the hippocampus. The following results were obtained. Methods : This study used 21 male specific pathogen free (SPF) SD (Sprague Dawley) rats, 8 weeks of age and approximately 300 g in weight, that were given at least 1 week to adapt to the lab environment Each exposed artery was completely occluded with non-absorbent suture thread and kept in that state for 5 minutes. The sutures were then removed to allow reperfusion of blood. Test group is control group for comparison with the common carotid artery occlusion models, a GI group that underwent common carotid artery occlusion, and a needle electrode electrical stimulation (NEES) group that underwent NEES after artery occlusion. The GI and NEES groups were given 12, 24, or 48 hours of reperfusion before NEES. NEES device (PG6, ITO, Japan, 9V, current, 2Hz) was used to stimulate the right and left acupoint ST36 of the SD rats for 30 minutes while they were sedated with 3% isoflurane. An immunohistochemistry test was done on the forebrains of the GI induced rats. All the data collected from this study was symbolized and analyzed using a statistics processing program (SPSS 12.0K/PC). The level of significance was set at ${\alpha}$=0.05 and a T-TEST analysis was used to find out the effects of treatment on each of the groups: the normal group, the CVA induced group, and the treatment after CVA induction group. Results : Both PARP and C-fos immuno-reactive cells, related to apoptosis, were greater in the GI groups than the NEES group. Caspase and IL-6 immuno-reactive cells, related to inflammation, were greater in the GI and NEES groups than the control group. Conclusions : This research was conducted to study the effects of NEES on CVA due to ischemia. Occlusion and reperfusion was performed on the common carotid arteries of white rats, after which amounts of substances related to neuron necrosis and inflammation - PARP, IL-6, Caspase-3, and C-fos - were measured in the Hippocampus

• 한국임상수의학회지
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• 제30권3호
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• pp.159-165
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• 2013

본 연구는 내측반월판 절제와 전십자인대 단열로 유발된 랫드 골관절염 모델을 이용하여 계피 (Cinnamomum cassia Blume, 육계(肉桂)) 추출물의 연골손상 방지에 대한 효과를 평가하였다. 골관절염유발 랫드 48마리를 군당 8마리씩 6군으로 음성 대조군과 골관절염 대조군, 체중당 diclofenac 2 mg 투여군, 계피 추출물 25 mg과 50 mg, 100 mg 투여군으로 각각 분류하여 수술 1주 후부터 12주동안 투여하였다. 연골 소실과 관절의 불안정성은 계피 추출물과 diclofenac 투여군이 골관절염 대조군과 비교할 때 유의하게 감소하였다(p < 0.05). 병리조직학적 평가에서 연골의 퇴행은 계피 추출물 투여량에 따라 용량 의존적으로 개선됨이 확인되었다(p < 0.01). 계피 추출물 투여군에서 관절구조 퇴행성 변화의 감소와 Safranin-O 염색 정도의 용량 의존적인 증가를 보여 연골 퇴행이 억제됨을 확인하였다. Diclofenac이 골관절염 진행에 있어 활성화된 caspase-3와 절단된 poly(ADP-ribose) 중합효소에 유도된 세포자멸사 표지율 증가에 별 다른 영향을 주지 않았지만 계피 추출물 투여군에서는 이들 세포자멸 표지자에 대한 반응세포는 유의하게 감소되었다(p < 0.05). 그러나, diclofenac과 계피 추출물 투여군은 5-bromo-2-deoxyuridine의 섭취에는 영향을 주지 않았다. 이러한 결과에서 계피추출물이 항염활성과 항세포 자멸 활성을 통해 관절연골에 보호 효과가 있음을 보였다.

• Nutrition Research and Practice
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• 제12권2호
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• pp.93-100
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• 2018

BACKGROUND/OBJECTIVE: Oxidative stress plays a key role in neuronal cell damage, which is associated with neurodegenerative disease. The aim of present study was to investigate the neuroprotective effects of perilla oil (PO) and its active component, alpha-linolenic acid (ALA), against hydrogen peroxide $(H_2O_2)$-induced oxidative stress in SH-SY5Y neuronal cells. MATERIALS/METHODS: The SH-SY5Y human neuroblastoma cells exposed to $250{\mu}M$ $H_2O_2$ for 24 h were treated with different concentrations of PO (25, 125, 250 and $500{\mu}g/mL$) and its major fatty acid, ALA (1, 2.5, 5 and $25{\mu}g/mL$). We examined the effects of PO and ALA on $H_2O_2$-induced cell viability, lactate dehydrogenase (LDH) release, and nuclear condensation. Moreover, we determined whether PO and ALA regulated the apoptosis-related protein expressions, such as cleaved-poly ADP ribose polymerase (PARP), cleaved caspase-9 and -3, BCL-2 and BAX. RESULTS: Treatment of $H_2O_2$ resulted in decreased cell viability, increased LDH release, and increase in the nuclei condensation as indicated by Hoechst 33342 staining. However, PO and ALA treatment significantly attenuated the neuronal cell death, indicating that PO and ALA potently blocked the $H_2O_2$-induced neuronal apoptosis. Furthermore, cleaved-PARP, cleaved caspase-9 and -3 activations were significantly decreased in the presence of PO and ALA, and the $H_2O_2$-induced up-regulated BAX/BCL-2 ratio was blocked after treatment with PO and ALA. CONCLUSIONS: PO and its main fatty acid, ALA, exerted the protective activity from neuronal oxidative stress induced by $H_2O_2$. They regulated apoptotic pathway in neuronal cell death by alleviation of BAX/BCL-2 ratio, and down-regulation of cleaved-PARP and cleaved caspase-9 and -3. Although further studies are required to verify the protective mechanisms of PO and ALA from neuronal damage, PO and ALA are the promising agent against oxidative stress-induced apoptotic neuronal cell death.