• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3293-3299
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• 2015

Background: Arm protein lost in epithelial cancers, on chromosome X (ALEX) is a novel subgroup within the armadillo (ARM) family, which has one or two ARM repeat domains as opposed to more than six-thirteen repeats in the classical Armadillo family members. Materials and Methods: In the study, we explore the biological functions of ALEX1 in breast cancer cells. Overexpression of ALEX1 and silencing of ALEX1 were performed with SK-BR3 and MCF-7 cell lines. Cell proliferation and colony formation assays, along with flow cytometry, were carried out to evaluate the roles of ALEX1. Results: ALEX1 overexpression in SK-BR3 breast cancer cells inhibited proliferation and induced apoptosis. Furthermore, depletion of ALEX1 in MCF-7 breast cancer cells increased proliferation and inhibited apoptosis. Additional analyses demonstrated that the overexpression of ALEX1 activated the intrinsic apoptosis cascades through up-regulating the expression of Bax, cytosol cytochrome c, active caspase-9 and active caspase-3 and down-regulating the levels of Bcl-2 and mitochondria cytochrome c. Simultaneouly, silencing of ALEX1 inhibited intrinsic apoptosis cascades through down-regulating the expression of Bax, cytosol cytochrome c, active caspase-9, and active caspase-3 and up-regulating the level of Bcl-2 and mitochondria cytochrome c. Conclusions: Our data suggest that ALEX1 as a crucial tumor suppressor gene has been involved in cell proliferation and apoptosis in breast cancer, which may serve as a novel candidate therapeutic target.

• 한국식품저장유통학회지
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• 제31권2호
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• pp.276-286
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• 2024

본 연구에서는 후코이단/이고들빼기 혼합물이 HepG2 세포의 apoptosis에 미치는 영향을 확인하고 어떠한 경로를 통해 나타나는지를 조사하였다. 후코이단/이고들빼기 혼합물이 HepG2 세포의 증식을 억제하고 세포 독성을 나타냈다. 이러한 HepG2 세포의 증식 억제 및 세포 독성이 apoptosis에 의한 효과인지를 확인한 결과, DNA fragmentation과 mitochondria membrane potential의 저해를 일으키는 것을 확인하였다. 이러한 결과를 바탕으로 HepG2 세포에서 후코이단/이고들빼기 혼합물이 apoptosis를 유도하는 기전에 관여하는 단백질의 발현 양상을 확인한 결과, intrinsic apoptosis 경로인 p53을 증가시키고, Bcl-2 family인 Bcl-2의 억제 및 BAX의 증가를 통해서 cytochrome c를 증가시켜 caspase-9 활성화하였고, caspase-3를 활성화시켜 결과적으로 apoptosis를 유도하였다. 또한, 전반적으로 후코이단/이고들빼기 혼합물의 apoptosis 유도 효과는 후코이단만 처리한 것보다 더 높은 효과를 나타냈으며, 이는 이고들빼기 추출물과의 혼합물 제조를 통해서 후코이단의 apoptosis 유도 효과가 증대되는 것으로 판단된다. 이러한 결과는 후코이단/이고들빼기 혼합물이 HepG2 세포에서 apoptosis 관련 유전자의 발현 조절에 의해 항암 효과를 나타내며, 이는 후코이단/이고들빼기 혼합물의 항암 작용의 기전을 해석하였을 뿐만 아니라, 이러한 기전 연구를 바탕으로 실질적인 간암 치료제로서의 사용 가능성을 확인하기 위해서는 유용 물질 분석 및 in vivo에서 추가 실험 등이 다양하게 수행되어야 할 것으로 판단된다.

• 한국수정란이식학회지
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• 제32권4호
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• pp.275-285
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• 2017

Lysophosphatidic acid (LPA) is an important signaling molecule. Here, the effect and mechanism of LPA on the preimplantation development of porcine embryos during in vitro culture (IVC) was examined. Porcine embryos were cultured in porcine zygote medium (PZM-3) supplemented with $30{\mu}M$ LPA during different days. There was a significantly higher cleavage rate in Day 1-7 and significantly higher total cell number of blastocysts in Day 1-3 and Day 4-7. It was also found that messenger RNA (mRNA) expression level of PCNA, BCL2 and BAX in blastocysts obtained from D1-7 group were significantly higher and BCL2/BAX mRNA ratio in D1-3 group was significantly lower than control group but Day 4-7 and Day 1-7 groups were comparable with control group. Treatment with $20{\mu}M$ PLC inhibitor significantly decreased the embryo cleavage rate and blastocyst formation rate. Moreover, LPA as an activator of PLCs, enhanced the $30{\mu}M$ LPA + $20{\mu}M$ U73122 group embryo cleavage rate which similar with control group. In conclusion, the results suggest that treatment with LPA during IVC improves the porcine early embryo cleavage by activation of PLC signaling pathway and regulate the mRNA expression that contribute to total cell number of blastocysts during blastocyst formation.

• The Korean Journal of Physiology and Pharmacology
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• 제23권1호
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• pp.21-28
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• 2019

Swertiamarin (STM) is an iridoid compound that is present in the Gentianaceae swertia genus. Here we investigated antiapoptotic effects of STM on carbon tetrachloride ($CCl_4$)-induced liver injury and its possible mechanisms. Adult male Sprague Dawley rats were randomly divided into a control group, an STM 200 mg/kg group, a $CCl_4$ group, a $CCl_4+STM$ 100 mg/kg group, and a $CCl_4+STM$ 200 mg/kg group. Rats in experimental groups were subcutaneously injected with 40% $CCl_4$ twice weekly for 8 weeks. STM (100 and 200 mg/kg per day) was orally given to experimental rats by gavage for 8 consecutive weeks. Hepatocyte apoptosis was determined by TUNEL assay and the expression levels of Bcl-2, Bax, and cleaved caspase-3 proteins were evaluated by western blot analysis. The expression of $TGF-{\beta}1$, collagen I, collagen III, CTGF and fibronectin mRNA were estimated by qRT-PCR. The results showed that STM significantly reduced the number of TUNEL-positive cells compared with the $CCl_4$ group. The levels of Bax and cleaved caspase-3 proteins, and $TGF-{\beta}1$, collagen I, collagen III, CTGF, and fibronectin mRNA were significantly reduced by STM compared with the $CCl_4$ group. In addition, STM markedly abrogated the repression of Bcl-2 by $CCl_4$. STM also attenuated the activation of the PI3K/Akt pathway in the liver. These results suggested that STM ameliorated $CCl_4$-induced hepatocyte apoptosis in rats.

• 대한한의학방제학회지
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• 제24권4호
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• pp.311-321
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• 2016

Chronic or acute alcohol abuse often leads to liver injury associated with alcoholic hepatitis, liver fibrosis, cirrhosis, and liver cancer. In addition to the liver, alcohol abuse also induces a variety of other tissue injuries including pancreatitis, cardiomyopathy, neurotoxicity and muscle loss. Chronic skeletal muscle myopathy, independent of peripheral neuropathy, is well recognised in alcoholic patients. Several mechanisms may be involved in the pathogenesis of alcoholic myopathy. Ethanol is a potent inhibitor of muscle protein synthesis. Gastrocnemius and plantaris muscles are Type II fiber-predominant and usually considered representative of the musculature as a whole. Whereas, soleus muscle is Type I fiber predominant. Shihosogan-san is a traditional Korean medicine that is widely employed to treat indigestion and liver diseases. Muscle diseases are often related to liver diseases and conditions. We therefore tested the hypothesis that treatment with Shihosogan-san could ameliorate the ethanol-induced changes in muscle protein synthesis. Young male Sprague-Dawley rats were orally given 25% ethanol (5ml/kg, body weight) daily with Ethanol for 28 days. Normal group was similarly administrated with saline. In Shihosogan-san treated group, rats were orally administrated Shihosogan-san extract, and rats of EtOH group were given with the vehicle only. After 4 week, the morphology of gastrocnemius and plantaris muscles were assessed by hematoxylin and eosin staining. For comparative purposes, liver function was also investigated. The muscles from rats of EtOH group displayed a significant reduction in average cross section area compared to Normal group. Shihosogan-san treated group had increased fiber compared to the EtOH group. Moreover, Shihosogan-san treated group compared with EtOH group showed significantly decreased pro-apoptotic BAX expression and increased anti-apoptotic Bcl-2 expression. In conclusion, Shihosogan-san extract showed ameliorating effects on chronic alcohol toxicity in skeletal muscle.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.21-26
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• 2011

Ceramide is an important lipid mediator of extracellular signals that control various cellular functions, including apoptosis. In this study, we showed that ceramide induced apoptosis in U373MG human glioblastoma cells associated with G1 cell cycle arrest. Treatment of cells with ceramide increased proapoptotic Bax expression and inhibited the expression of antiapoptotic Bcl-2 and Bcl-xL Ceramide also downregulated cyclin E, cyclin D1, cdk 2, and cdk4 which are involved in regulating cell cycle. In addition, ceramide suppressed phosphorylation of Akt, Bad, p70 S6 kinase, and 4E-BP1, suggesting the involvement of Akt/mTOR signaling pathway. Additionally, okadaic acid, an inhibitor of protein phosphatase 2A, partially blocked the ceramide mediated inhibition of phosphorylation of Akt and 4E-BP1. These results suggest that ceramide induces apoptosis in U373MG glioblastoma cells by regulating multiple signaling pathways that involve cell cycle arrest associated with Akt signaling pathway.

• 한국자원식물학회지
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• 제31권3호
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• pp.268-273
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• 2018

Hepatic ischemia-reperfusion injury (HIRI) is linked with high mortality rate. Several agents have been developed so far to reduce the risk of HIRI. In this study, we investigated the effects of combined treatment of Ginko biloba leaves extract and vitamin C (GLEVC) on hepatic ischemia-reperfusion injury. To explore the protective effects of GLEVC on HIRI rats model were tested. After the development of HIRI by using clamping method rats were then randomly divided into four groups. Different doses of GLEVC were administered in HIRI rat model. The level of ALT, AST, SOD and MDA content in serum were detected in HIRI groups. Moreover, the activity of SOD, content of MDA, and GSH in hepatic tissue were also examined. Bcl-2 and Bax protein expression were detected by immunohistochemical staining method. Compared with sham group, GLEVC has the protective effect on the HIRI-induced model. Level of ALT, AST, and MDA in blood were significantly lower in GLEVC group compared with HIRI-induced group. Moreover, SOD activity and GSH were increased in GLEVC group whereas MDA content was reduced by GLEVC treatment. Furthermore, HIRI-induced Bax protein was reduced upon GLEVC treatment, whereas Bcl-2 protein expression was enhanced. These results demonstrate that GLEVC treatment may provide potential ameliorative therapy by reducing damaged signaling mechanism in hepatic ischemia/reperfusion injury model.

• Journal of Pharmaceutical Investigation
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• 제37권5호
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• pp.287-290
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• 2007

Intrinsic or acquired resistance to chemotherapeutic drugs is one of the major obstacles to effective cancer treatment. Hypoxia is widespread in solid tumors as a consequence of decreased blood flow in the tumor-derived neovasculature. The recent finding of a link between hypoxia and chemoresistance prompted us to investigate whether hypoxia induces doxorubicin resistance in human MCF-7 breast cancer cells. Low oxygen concentration decreased the doxorubicin sensitivity in MCF-7 cells. The expression of p-glycoprotein, a major MDR-related transporter, and those of apoptosis-related proteins (anti-apoptotic Bcl-2, Bcl-XL and pro-apoptotic Bax) were not altered by hypoxia in MCF-7 cells. Intracellular uptake of doxorubicin was significantly decreased under hypoxic conditions. Decreased cellular uptake of doxorubicin under hypoxia may contribute to causing doxorubicin resistance in these cells. The use of agents that can modulate the doxorubicin uptake for adjuvant therapy may contribute to improving the therapeutic efficacy of doxorubicin in breast cancer patients.

• 한국식품영양과학회지
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• 제45권11호
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• pp.1589-1594
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• 2016

본 연구는 후추과 식물의 주성분인 piperine이 위암세포 AGS의 항암 효과에 미치는 영향을 확인하기 위해 수행되었다. Piperine에 의한 위암세포 AGS의 생존율을 확인하기 위해 MTT assay를 수행한 결과 농도 의존적으로 암세포의 생존율이 감소하는 것을 확인하였다. 이러한 암세포의 생존율 억제가 apoptosis에 의한 효과인지를 확인하기 위해 DAPI staining을 실시한 결과 piperine을 처치한 군에서 apoptotic body와 염색질 응축이 증가하는 것을 확인하였다. MTT assay와 DAPI staining의 결과를 바탕으로 piperine이 위암세포 AGS에서 apoptosis와 관련한 단백질 발현 양상에 미치는 영향을 확인하기 위해 western blotting을 실시하였다. 그 결과 piperine에 의해 AGS 세포에서 apoptosis를 유도하는 p53, Bax의 발현은 농도 의존적으로 증가하였고, apoptosis를 억제하는 Bcl-2, XIAP의 발현은 농도 의존적으로 감소하였다. Apoptosis의 궁극적 단계인 caspase-9와 손상된 DNA를 복구하는 PARP의 분절은 증가하였으며, apoptosis를 방해하는 인자인 Akt의 인산화는 농도 의존적으로 감소하였다. Akt 억제제인 LY294002와 piperine을 병행 또는 단독으로 처치하여 western blotting을 진행한 결과 Bcl-2의 발현은 piperine을 단독으로 처치했을 때와 유사한 감소 경향을 보였지만, p-Akt의 발현은 대조군과 비교해 LY294002와 piperine을 단독으로 처치했을 때보다 병행했을 때 더욱 감소하였다. 이와 반대로 Bax와 cleaved-PARP의 발현은 강하게 증가하였다. 본 연구의 결과를 종합해볼 때 piperine이 위암세포 AGS에서 Akt 경로를 통해 apoptosis를 유도하는 것으로 여겨지며, 향후 위암 예방제나 치료제로의 개발 가능성이 있을 것으로 생각한다.

• Clinical and Experimental Pediatrics
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• 제51권3호
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• pp.307-314
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• 2008

목 적 : 한국인 음식에 널리 이용되고 있는 중요한 향신료인 고추에 많이 들어있는 capsaicin이 최근 많은 연구를 통해 암을 예방하고 더 나아가 치료할 수 있는 성분으로 밝혀지고 있다. 따라서 본 연구의 목적은 한국인이 많이 섭취하고 있는 식이성분인 capsaicin이 한국인에 많은 위암세포의 세포자멸사의 유도물질로 가능성을 알아보고 이를 항암치료의 일부분으로 시도하고자 하였다. 방 법 : 한국인의 위암세포주인 SNU-668 세포에 capsaicn를 처리한 후 세포 생존은 trypan blue 와 crystal violet 분석, 세포독성은 MTT 분석, 세포사 분석은 핵 응축과 DNA 분절화 실험, bcl-2와 bax의 mRNA 발현은 역전사 중합효소 연쇄반응, 세포사와 관련된 단백질 발현은 Western immunoblot analysis로 분석하였다. Capsaicin에 의해 항암제의 감수성이 증가되는 지를 알아보기 위하여 일정 농도의 capsaicin과 농도 별 각 항암제를 같이 처리하여 2일 간 배양 하고 MTT assay로 분석하였으며 이와 관련된 세포사와 관련된 단백질 발현을 분석하였다. 결 과 : capsaicin의 농도에 따라 SNU-668 위암세포의 증식을 현저히 억제하였으며, 이러한 capsaicin의 증식 억제는 DNA 단편화, 핵 응축과 caspase 활성화에 의해 세포자멸사의 유도에 의한 것으로 입증되었다. 더욱이 capsaicin에 의해 pro-apoptotic Bax에 대한 anti-apoptotic Bcl-2의 비율이 현저히 감소하고, caspase-3활성이 증가하였다. Capsaicin을 처리한 세포는 처리하지 않은 세포에 비하여 etoposide나 adriamycin에 의해 유도되는 세포사멸에 더욱 감수성을 보였다. 결 론 : 이상의 연구에서 capsaicin은 caspase-3 의존적 경로를 통하여 SNU-668 세포에서 세포자멸사를 유도하며 항암제의 감수성을 증가시키므로 위암의 효과적인 항암치료제와 항암제 민감성 유도제의 일환으로 가능성이 있다.