• BMB Reports
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• 제42권11호
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• pp.719-724
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• 2009

Recent studies have revealed that endoplasmic reticulum (ER) disturbance is involved in the pathophysiology of neurodegenerative disorders, contributing to the activation of the ER stress-mediated apoptotic pathway. Therefore, we investigated here the molecular mechanisms underlying the ER-mitochondria axis, focusing on calcium as a potential mediator of cell death signals. Using NT2 cells treated with brefeldin A or tunicamycin, we observed that ER stress induces changes in the mitochondrial function, impairing mitochondrial membrane potential and distressing mitochondrial respiratory chain complex Moreover, stress stimuli at ER level evoked calcium fluxes between ER and mitochondria. Under these conditions, ER stress activated the unfolded protein response by an overexpression of GRP78, and also caspase-4 and-2, both involved upstream of caspase-9. Our findings show that ER and mitochondria interconnection plays a prominent role in the induction of neuronal cell death under particular stress circumstances.

• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.3865-3869
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• 2016

Curcumin, a polyphenolic compound isolated from the rhizomes of an herbaceous perennial plant, Curcuma longa, is known to possess anticancerous activity. However, the mechanism of apoptosis induction in cancers differs. In this study, we have (1) investigated the anticancerous activity of curcumin on REH and RS4;11 leukemia cells and (2) studied the chemo-sensitizing potential of curcumin for doxorubicin, a drug presently used for leukemia treatment. It was found that curcumin induced a dose dependent decrease in cell viability because of apoptosis induction as visualized by annexin V-FITC/ PI staining. Curcumin-induced apoptosis of leukemia cells was mediated by PARP-1 cleavage. An increased level of caspase-3, apoptosis inducing factor (AIF), cleaved PARP-1 and decreased level of Bcl2 was observed in leukemia cells after 24h of curcumin treatment. In addition, curcumin at doses lower than the $IC_{50}$ value significantly enhanced doxorubicin induced cell death. Therefore, we conclude that curcumin induces apoptosis in leukemia cells via PARP-1 mediated caspase-3 dependent pathway and further may act as a potential chemo-sensitizing agent for doxorubicin. Our study highlights the chemo-preventive and chemo-sensitizing role of curcumin.

• 생명과학회지
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• 제31권11호
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• pp.969-979
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• 2021

당뇨병은 만성대사성 질환으로 우리나라 국민의 사망요인 중 5위를 차지하고 있다. 제 2형 당뇨병에서 나타나는 인슐린 분비 감소는 베타세포의 자가사멸에 의한 베타세포질량의 급격한 감소로 인한 것으로 보고되고 있으며, 베타세포의 자가사멸을 촉진하는 요인으로 고혈당에 의한 당독성 및 활성산소종들의 증강에 의한 산화스트레스 등이다. 벼메뚜기 추출물은 고농도 포도당 처리된 INS-1 췌장 베타 세포에서 세포 생존율을 증가시키고, 지질과산화물, 세포 내 ROS 및 NO 수준을 감소시켰다. 세포사멸 관련 인자의 유전자 발현 결과 pro-세포자가사멸인자인 Bax, Cytochrome C, caspase-3 및 caspase-9의 단백질 발현을 유의적으로 감소시켰고, anti-세포자가사멸인자인 Bcl-2 발현을 증가시켰다. Annexin V/I propidium iodide 염색법을 통하여 벼메뚜기 추출물이 고농도 포도당으로 유도된 세포 사멸을 감소시키고, INS-1 췌장 베타세포에서의 인슐린 분비능을 증가시키는 것으로 사료된다. 그러므로 벼메뚜기 추출물이 고농도 포도당으로 손상된 INS-1 췌장 베타세포의 보호효과를 나타낸다. 본 연구의 결과는 벼메뚜기 추출물이 당뇨병 치료에 도움을 주는 항당뇨 기능성 식품 소재 및 개발에 기여할 수 있음을 시사한다.

• Clinical and Experimental Pediatrics
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• 제46권7호
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• pp.702-709
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• 2003

목 적 : $IFN{\gamma}$는 다양한 암세포에서 $TNF{\alpha}$와 FAS/CD95 수용체 발현을 증가시키거나 caspase나 Bcl-2 가족의 활성화를 조절하여 $TNF{\alpha}$와 FAS/FASL 유도성 세포고사를 촉진한다. 신경모세포종에서 $IFN{\gamma}$와 $TNF{\alpha}$는 협동적으로 세포 분화를 유도하거나 성장 억제를 일으킨다. 또한 일부 신경모세포종에서 자연적인 FAS 수용체 발현에도 불구하고 그 리간드 자극에 의한 세포고사 유도에는 실패하였고 $IFN{\gamma}$ 투여로 이를 극복할 수 있음이 보고되었다. 본 연구에서는 $IFN{\gamma}$가 $TNF{\alpha}$나 길항적 FAS/CD95 항체 유도성 세포고사를 촉진할 수 있는지 여부를 다양한 항암제에 대한 내성을 가지고 있는 신경모세포종 세포주를 이용하여 알아보았다. 방 법 : CHLA-15, CHLA-90와 LA-N-2 신경모세포종 세포주를 IMDM 배지로 배양하였고 유전자 재조합 $IFN{\gamma}$, $TNF{\alpha}$, 길항적 FAS/CD95 항체(CH-11)를 투여하였다. 세포 생존율은 형광기질인 calcein-AM을 이용한 DIMSCAN을 통하여 측정하였고, 세포고사 정도는 Annexin V-PE와 7-ADD염색을 이용한 유식세포 분석기를 통하여 분석하였고 pancaspase and caspase-8 억제 실험을 통하여 확인하였다. TNF와 FAS/CD95 수용체 표현은 각각에 대한 단클론 항체와 PE가 결합된 이차 항체를 이용하여 유식세포 분석기로 알아보았다. 결 과 : $IFN{\gamma}$ 또는 $TNF{\alpha}$ 단독 투여로는 모든 세포주에서 의의있는 세포 독성을 유도하지 못 했으나 $IFN{\gamma}$와 $TNF{\alpha}$을 병행 투여시에는 CHLA-15과 CHLA-90 세포주에서 의의있는 세포 생존율 감소와 공통 capase경로를 통한 세포고사를 협동적으로 촉진하였다. 또한 길항적 FAS/CD95 항체 단독 투여 시에는 모든 세포주에서 세포 생존율의 변화가 없었으나 $IFN{\gamma}$ 전 처치 후 투여 시에는 CHLA-90 세포주에서 현저한 세포 생존율 변화 및 세포고사를 유도하였다. $INF{\gamma}$ 치료 후 TNFRI와 FASR의 발현이 모든 세포주에서 현저히 증가하였는데 이는 일부 감수성이 있는 신경모세포종에서 $INF{\gamma}$에 의한 $TNF{\alpha}$와 FAS/CD95수용체 유도성 세포고사 촉진의 한 기전이 될 것으로 사료된다. 결 론: 일부 신경모세포종에서 $IFN{\gamma}$가 $TNF{\alpha}$와 길항적 FAS/CD95 항체 유도성 세포고사를 감작화 시켰으며 이는 수용체 발현의 증가와 동반되었다.

• 대한의생명과학회지
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• 제19권2호
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• pp.118-123
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• 2013

Tumor necrosis factor-alpha (TNF-${\alpha}$) is a proinflammatory cytokine that mediates the inflammatory response and immune functions, and modulates the proliferation, differentiation and cell death of cancer cells. The differential functions of TNF-${\alpha}$ in various human cells due to the formation of different stimulating pathway upon the binding of TNF-${\alpha}$ to its receptors. In the present study, we examined the different effects of TNF-${\alpha}$ on the survival and apoptosis between normal granulocytes and human myeloid leukemia HL-60 cells. Although TNF-${\alpha}$ did not affect on the constitutive apoptosis of granulocytes, TNF-${\alpha}$ strongly induced the apoptosis of HL-60 cells in a dose- and a time-dependent manner. TNF-${\alpha}$-induced apoptosis was occurred via the activation of caspase 8, caspase 9 and caspase 3/7 and the induction of ROS production in HL-60 cells. Also, BAY-11-7085, a NF-${\kappa}B$ inhibitor, blocked the TNF-${\alpha}$-induced apoptosis in HL-60 cells. NF-${\kappa}B$ may be involved in TNF-${\alpha}$-induced apoptotic signaling pathway in HL-60 cells. These results suggest that TNF-${\alpha}$ activates apoptotic pathways and its process depends on cell type and many cellular factors. A better understanding of the differential effect of TNF-${\alpha}$ on cell apoptosis and survival may provide important information that can be used to elucidate the specific inhibitory effect of TNF-${\alpha}$ on the cancer dis.

• Journal of Microbiology and Biotechnology
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• 제18권11호
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• pp.1862-1867
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• 2008

Streptochlorin is a small molecule isolated from marine Streptomyces sp. that is known to have antiangiogenic and anticancer properties. In this study, we examined the effects of this compound on reactive oxygen species (ROS) production and the association of these effects with apoptotic tumor cell death, using a human hepatocarcinoma Hep3B cell line. The results of this study demonstrated that streptochlorin mediates ROS production, and that this mediation is followed by a decrease in the mitochondrial membrane potential (MMP, ${\Delta}{\Psi}_m$), activation of caspase-3, and downregulation of antiapoptotic Bcl-2 protein. The quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the streptochlorin-induced apoptosis effects via inhibition of ROS production, MMP collapse, and the subsequent activation of caspase-3. These observations clearly indicate that ROS are involved in the early molecular events in the streptochlorin-induced apoptotic pathway. Taken together, our data imply that streptochlorin-induced ROS is a key mediator of MMP collapse, which leads to the caspase-3 activation, culminating in apoptosis.

• 동의생리병리학회지
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• 제20권1호
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• pp.187-192
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• 2006

Herbal medicines have been utilized to treat a variety of diseases, including cancer. Several species of the family rubiaceae have been reported to have antitumor activity. In this study, we report the cytotoxicity and antitumor activity exhibited dy the methanol extracts prepared from Rubia radix (RRME), Uncaria gambir (UGME) and Oldenlandia diffusa (ODME) (family: Rubiaceae) against human promyleloid leukemia cell line, HL-60. The cytotoxicity of RRME (2~20 ${\mu}g/ml$), UGME (20~200 ${\mu}g/ml$) and ODME (20~200 ${\mu}g/ml$) were assessed dy the MTT reduction assay. IC50 values for RRME, UGME and ODME were 11.0, 99.5 and 106.1 ${\mu}g/ml$, respectively. When the HL-60 cells were treated with RRME (10 ${\mu}g/ml$), UGME (120 ${\mu}g/ml$) and ODME (140 ${\mu}g/ml$) for 24 h, several apoptotic characteristics such as DNA fragmentation and morphologic changes were observed. Furthermore, flow cytometric analysis was peformed to determine the percent of apoptotic cells. The poupulation of sub-G1 hypodiploid cells was increased 37.49% in RRME treatment, 12.49% in UGME treatment and 7.21% in ODME treatment compared with untreated control cells (2.64%). To further confirm apoptotic cell death, we assayed caspase-3, -8 and -9 activities in RRME, UGME and ODME-treated cells. After treatment of RRME, UGME and ODME for 12 h, caspase-3, -8 and -9 activities significantly increased.compared to untreated control cells. These results show that RRME, UGME and ODME induced apoptotic cell death in HL-60 cells and may have a possibility of potential antitumor activities.

• Journal of Ginseng Research
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• 제47권3호
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• pp.400-407
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• 2023

Background: Rb3 is a ginsenoside with anti-inflammatory properties in many cell types and has been reported to attenuate inflammation-related metabolic diseases such as insulin resistance, nonalcoholic fatty liver disease, and cardiovascular disease. However, the effect of Rb3 on podocyte apoptosis under hyperlipidemic conditions, which contributes to the development of obesity-mediated renal disease, remains unclear. In the current study, we aimed to investigate the effect of Rb3 on podocyte apoptosis in the presence of palmitate and explore its underlying molecular mechanisms. Methods: Human podocytes (CIHP-1 cells) were exposed to Rb3 in the presence of palmitate as a model of hyperlipidemia. Cell viability was assessed by MTT assay. The effects of Rb3 on the expression of various proteins were analyzed by Western blotting. Apoptosis levels were determined by MTT assay, caspase 3 activity assay, and cleaved caspase 3 expression. Results: We found that Rb3 treatment alleviated the impairment of cell viability and increased caspase 3 activity as well as inflammatory markers in palmitate-treated podocytes. Treatment with Rb3 dosedependently increased PPARδ and SIRT6 expression. Knockdown of PPARδ or SIRT6 reduced the effects of Rb3 on apoptosis as well as inflammation and oxidative stress in cultured podocytes. Conclusions: The current results suggest that Rb3 alleviates inflammation and oxidative stress via PPARδ-or SIRT6-mediated signaling, thereby attenuating apoptosis in podocytes in the presence of palmitate. The present study provides Rb3 as an effective strategy for treating obesity-mediated renal injury.

• 생명과학회지
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• 제19권11호
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• pp.1581-1590
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• 2009

H. fusiformis는 갈조식물, 모자반과의 해조류로서 우리나라의 서해안, 남해안, 및 제주도를 비롯해 중국, 일본 등 아시아 전역에 서식하는 천연자원식물이다. 최근 톳의 항산화 활성에 대한 연구는 활발히 이루어지고 있으나 항암에 대한 연구 및 분자생물학적 기전연구가 많이 미흡한 편이다. 따라서 본 연구에서는 인체 유방암세포주인 MDA-MB-231 및 MCF-7 세포를 대상으로 톳 에탄올 추출물(EAHF)에 의한 증식억제에 대하여 조사한 결과, 두 세포주 모두에서 EAHF을 처리하지 않은 군과 비교하여 EAHF의 처리 농도가 증가할수록 암세포의 증식이 현저하게 억제되었고 심한 형태적 변화를 관찰할 수 있었다. 이러한 유방암 세포의 증식억제 및 형태변화는 MDAMB-231에서는 apoptosis 유발과 밀접한 관련이 있음을 알 수 있었고, EAHF에 의해 유발된 apoptosis 정도를 정량적으로 분석하기 위하여 flow cytometry를 이용하여 apoptosis 유발 세포군에 해당하는 sub-G1기에 속하는 세포들의 빈도를 측정하였다. MDA-MB-231세포의 경우는 EAHF의 처리 농도의 증가에 따라 sub-G1기에 해당하는 세포의 빈도가 증가하여 $400{\mu}g/ml$의 농도에서는 약 23% 정도가 나타났지만 MCF-7 세포에서는 apoptosis 유발보다는 G1 arrest 효과가 높게 나타났다. 또한 염색질 응축과 연관된 apoptotic body 형성과 함께 apoptosis 유발의 직접적인 증거에 해당하는 DNA fragmentation 여부를 agarose gel 전기영동으로 조사하였다. MDAMB-231 세포에서는 apoptosis가 일어난 세포들에서 볼 수 있는 전형적인 DNA laddering을 관찰 할 수 있었지만 MCF-7세포에서는 DNA laddering이 거의 관찰되지 않았다. 또한 apoptosis 유발에 관여하는 여러 유전자들의 발현에 EAHF이 어떠한 영향을 미치는지를 조사한 결과, MDA-MB-231 및 MCF-7 세포 모두에서 death receptor에 속하는 여러 유전자의 발현에는 아무런 변화가 나타나지 않았으나, MDA-MB-231 세포에서 pro-apoptotic Bax의 발현 증가와 caspase-9 및 -3의 활성화, caspase-3의 기질 단백질들의 분해 등이 관찰되었다.

• 대한의생명과학회지
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• 제20권4호
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• pp.244-249
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• 2014

Arazyme is a metalloprotease secreted by Aranicola proteolyticus that was previously shown to suppress cytokine expression of keratinocytes and endothelial cells and inhibit histopathological features in an atopic dermatitis-like animal model. However, the regulatory effects of arazyme in other allergic diseases have yet to be elucidated. In this study, we investigated whether arazyme is effective against neutrophil apoptosis in allergic diseases such as allergic rhinitis and asthma. Arazyme inhibited neutrophil apoptosis of normal subjects in a dose-dependent manner. However, the antiapoptotic effect of arazyme was reversed by LY294002, an inhibitor of PI3K, AKTi, an inhibitor of Akt, PD98059, an inhibitor of MEK, and BAY-11-7085, an inhibitor of NF-${\kappa}B$. Arazyme induced activation of NF-${\kappa}B$ via PI3K/Akt/ERK pathway. The anti-apoptotic effect of arazyme is associated with inhibition of cleavage of caspase 3 and caspase 9. Arazyme inhibited constitutive apoptosis of neutrophil in a dose-dependent manner in allergic subjects, and its mechanism was shown to be associated with PI3K/Akt/ERK/NF-${\kappa}B$. The results presented here improve our understanding of neutrophil apoptosis regulation and will facilitate development of drugs for treatment of allergic diseases.