• Journal of Food Hygiene and Safety
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• v.28 no.1
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• pp.41-44
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• 2013

Although tongcao has been reported to have anti-oxidative, anti-inflammatory and antipyretic effects, there is no report of the chemopreventive effect of tongcao in cancer cells. In the present study, we investigated the anti-proliferative activity of methanol extract of tongcao (MET) and its molecular target in HEp-2 human cervical cancer cells using MTS assay, western blot analysis, and DAPI staining. MET significantly decreases cell viability and induces apoptotic cell death. It affects Bid protein to be truncated resulting in the release of cytochrome c from mitochondria to cytosol whereas it did not affect other Bcl-2 family members. Thus, we clearly suggest that tongcao can be a potential naturally occurring plants having chemopreventive activity in cervical cancer.

• Journal of Acupuncture Research
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• v.31 no.2
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• pp.87-98
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• 2014

Objectives : We investigated whether snake venom toxin(SVT) from Vipera lebetina turanica sensitizes HT29 human epithelial colorectal cancer cells to tumor necrosis factor(TNF)-related apoptosis-inducing ligand(TRAIL) induced apoptosis in cancer cells. Methods : Cell viability assay was used to assess the inhibitory effect of TRAIL on cell growth of HT29 human colorectal cancer cells. And 6-diamidino-2-phenylindole(DAPI), terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay(TUNEL) staining assay were used to evaluate cell-apoptosis. Western blot analysis were conducted to observe apoptosis related proteins and death receptor. To assess whether the synergized inhibitory effect of SVT and TRAIL on reactive oxygen species(ROS) generation was reversed by strong anti-oxidative agent. Results : SVT with TRAIL inhibited HT29 cell growth different from TRAIL alone. Consistent with cell growth inhibition, the expression of TRAIL receptors; Expression of death receptor(DR)4 and DR5 was significantly increased and intrinsic pro-apoptotic cleaved caspase-3, -9 was subsequently increased together with increase of Bax/Bcl-2 ratio and extrinsic pro-apototic caspase-8 was also activated. In addition, the expression of anti-apoptotic survival proteins, a marker of TRAIL resistance(eg, cFLIP, survivin, X-linked inhibitor of apoptosis protein(XIAP) and Bcl-2) was suppressed by the combination treatment of SVT and TRAIL. Pretreatment with the ROS scavenger N-acetylcysteine abolished the SVT and TRAIL-induced upregulation of DR4 and DR5 expression and expression of the intrinsic pro-apoptotic caspase-3 and-9. Conclusion : The collective results suggest that SVT facilitates TRAIL-induced apoptosis in $HT_{29}$ human epithelial colorectal cancer cells through up-regulation of the TRAIL receptors; DR4 and DR5 and consecutive induction of bilateral apoptosis via regulating apoptosis related proteins.

• Journal of Acupuncture Research
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• v.20 no.5
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• pp.93-106
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• 2003

Objective : To investigate the possible molecular mechanism(s) of melittin as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Methods: MTT, morphological changes, DAPI staining, Western blot, RT-PCR and in vitro prostaglandin E2 (PGE2) accumulation assays were performed. Results: The anti-proliferative effect by melittin treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. Melittin induced apoptotic cell death in a concentration-dependent manner, which was associated with inhibition or degradation of apoptotic target proteins such as ${\beta}$-catenin, poly(ADP-ribose) polymerase(PARP) and phospholipase $C-{\gamma}1(PLC-{\gamma}1)$. Melittin treatment inhibited the expression of cyclooxygenase-2(COX-2) and accumulation of PGE2 in aconcentration-dependent fashion. In addition, Melittin treatment induced the down-regulation of telomerase reverse transcriptase(hTERT) and proto-oncogene c-myc expression of A549 cells. Conclusions: Taken together, these findings suggest that melittin-induced inhibition of human lung cancer cell proliferation is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and melittin may have therapeutic potential in human lung cancer.

• Journal of Acupuncture Research
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• v.28 no.3
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• pp.101-110
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• 2011

목적 : 이 연구는 봉독이 NF-${\kappa}$B의 활성억제를 통하여 전립선 암세포주인 DU-145 세포의 성장을 억제하 는지를 확인하고 그 기전을 살펴보고자 하였다. 방법 : 봉독을 처리한 후 DU-145의 성장억제를 관찰하기 위해 WST-1 assay를 시행하였고, 세포자멸사의 관찰에는 DAPI staining assay를 통한 세포형태관찰을 시행하였으며, 염증관련유전자 발현 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}$B의 활성 변화를 관찰하기 위해 EMSA와 luciferase assay를 시행하였으며, DU-145에서 봉독과 NF-${\kappa}$B의 상호작용을 관찰하기 위해 transient transfection assay를 시행하여 세포생존율과 NF-${\kappa}$B의 활성 변동을 측정하였다. 결과 : DU-145 세포에서 봉독을 처리한 후 세포성장이 억제되었으며, 염증관련유전자 발현 및 NF-${\kappa}$B의 활성의 유의한 감소를 나타내었다. DU-145 세포에서 NF-${\kappa}$B의 p50와 IKK들을 치환하여 작용기를 없애고 봉독을 처리하였을 경우에도 세포활성 및 NF-${\kappa}$B의 활성의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 봉독이 NF-${\kappa}$B의 활성 억제를 통하여 인간 전립선암세포주인 DU-145의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것으로 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다. 다만 그 기전에서 봉독은 기존연구와 같은 NF-${\kappa}$B p50 및 IKK들의 작용기와 상호작용 이외에 다른 기전이 관여되는 것으로 심화 연구를 요한다.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.6
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• pp.3855-3859
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• 2013

Purpose: Liver cancer, one of the most common cancers in China, is reported to feature relatively high morbidity and mortality. Curcumin (Cum) is considered as a drug possessing anti-angiogenic, anti-inflammation and anti-oxidation effect. Previous research has demonstrated antitumor effects in a series of cancers. Materials and Methods: In this study the in vitro cytotoxicity of Cum was measured by MTT assay and pro-apoptotic effects were assessed by DAPI staining and measurement of caspase-3 activity. In vivo anti-hepatoma efficacy of Cum was assessed with HepG2 xenografts. Results: It is found that Cum dose-dependently inhibited cell growth in HepG2 cells with activation of apoptosis. Moreover, Cum delayed the growth of liver cancer in a dose-dependent manner in nude mice. Conclusions: Cum might be a promising phytomedicine in cancer therapy and further efforts are needed to explore this therapeutic strategy.

• Journal of Life Science
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• v.23 no.6
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• pp.832-837
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• 2013

In this study, it was investigated the possible mechanisms by which glutamine deprivation exerts its anti-proliferative action in cultured human prostate carcinoma PC3 cells. Glutamine deprivation resulted in inhibition of growth and G2/M arrest of the cell cycle in a time-dependent manner without apoptosis induction, as determined by MTT assay, DAPI staining and flow cytometry analyses. The induction of G2/M arrest by glutamine deprivation was associated with the inhibition of expression of Cdc2, cyclin A and cyclin B1, and up-regulation of the expression of cyclin-dependent kinase (Cdk) inhibitor p21(WAF1/CIP1) in both transcriptional and translational levels. Moreover, glutamine deprivation increased the phosphorylation of checkpoint kinase (Chk)1 and Chk2; however, the levels of Cdc25C phosphorylation were decreased in response to glutamine deprivation in a time-dependent manner. Our data provide a first biochemical evidence that glutamine deprivation suppresses cell viability through G2/M phase arrest without induction of apoptosis in PC3 cells.

• Journal of Nutrition and Health
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• v.48 no.5
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• pp.451-456
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• 2015

Purpose: Neuronal apoptotic events induced by aging and hypoxic/ischemic conditions is an important risk factor in neurodegenerative diseases such as ischemia stroke and Alzheimer's disease. The peel of Citrus sunki Hort. ex Tanaka has long been used as a traditional medicine, based on multiple biological activities including anti-oxidant, anti-inflammation, and anti-obesity. In the current study, we examined the actions of fermented C. sunki peel extract against cobalt chloride ($CoCl_2$)-mediated hypoxic death in human neuroblastoma SH-SY5Y cells. Methods: Cell viability was measured by trypan blue exclusion. Expression of apoptosis related proteins and release of cytochrome c were detected by western blot. Production of intracellular reactive oxygen species (ROS) and apoptotic morphology were examined using 2',7'-dichlorofluorescin diacetate (DCF-DA) and 4',6-diamidino-2-phenylindole (DAPI) staining. Results: Exposure to $CoCl_2$, a well-known mimetic agent of hypoxic/ischemic condition, resulted in neuronal cell death via caspase-3 dependent pathway. Extract of fermented C. sunki peel significantly rescued the $CoCl_2$-induced neuronal toxicity with the cell viability and appearance of apoptotic morphology. Cytoprotection with fermented C. sunki peel extract was associated with a decrease in activities of caspase-3 and cleavage of poly (ADP ribose) polymerase (PARP). In addition, increase in the intracellular ROS and release of cytochrome c from mitochondria to the cytosol were inhibited by treatment with extract of fermented C. sunki peel. Conclusion: Based on these data, fermented C. sunki peel extract might have a protective effect against $CoCl_2$-induced neuronal injury partly through generation of ROS and effectors involved in mitochondrial mediated apoptosis.

• Journal of Acupuncture Research
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• v.18 no.3
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• pp.69-78
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• 2001

목적 : 본 연구는 최근 임상에서 많이 사용하는 자하차(紫河車) 약침액(藥鍼液)이 과산화수소($H_2O_2$)로 야기된 송과선 세포의 apoptosis에 있어서 세포 보호에 미치는 영향과 그 기전을 분석하였다. 방법 및 결과 : MTT assay를 이용하여 분석한 결과 처리 시간 및 농도에 따른 세포 독성의 효과가 $H_2O_2$ 투여로부터 관찰되었다. 또한 자하거 약침액은 세포 증식 효과를 나타내었고 자하거 약침액을 전처치하고 $H_2O_2$를 처치하였을 때 세포 독성이 크게 감소되었다. DAPI staining에서 자하거 약침액 투여군은 $H_2O_2$ 투여군에 비해 fragmentation이 억제되었다. TUNEL assay를 통하여 자하거 약침액 투여군은 $H_2O_2$ 투여군에 비하여 TUNEL 양성세포의 발현이 감소되었다. Flow cytometry를 통하여 자하거 약침액 투여군은 $H_2O_2$ 투여군에 비하여 세포주기 중 sub G1 분획의 증가가 억제되었다. 결론 : 이상의 결과를 통하여 자하거 약침액이 $H_2O_2$로 유발된 apoptosis에서 세포보호 효과가 있음이 확인되었다.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.38
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• pp.17.1-17.6
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• 2016

Background: The aim of this study is to verify the feasibility of using silk fibroin (SF) as a potential membrane for guided bone regeneration (GBR). Methods: Various cellular responses (i.e., cell attachment, viability, and proliferation) of osteoblast-like MG63 cells cultured on an SF membrane were quantified. After culturing on an SF membrane for 1, 5, and 7 days, the attachment and surface morphology of MG63 cells were examined by optical and scanning electron microscopy (SEM), cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and cell proliferation was quantified using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining. Results: Optical microscopy revealed that MG63 cells cultured on the SF membrane proliferated over the 7-day observation period. The viability of cells cultured on SF membranes (SF group) and on control surfaces (control group) increased over time (P < 0.05); however, at respective time points, cell viability was not significantly different between the two groups (P > 0.05). In contrast, cell proliferation was significantly higher in the SF membrane group than in the control group at 7 days (P < 0.05). Conclusions: These results suggest that silk fibroin is a biocompatible material that could be used as a suitable alternative barrier membrane for GBR.

• CELLMED
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• v.2 no.3
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• pp.26.1-26.5
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• 2012

Adverse side-effects and lack of scientific validation of some chemotherapeutic agents prevent the use of many traditional medicines claimed to have anti-cancer effects. Ethanolic extract of Conium maculatum has long been used in traditional and alternative systems of medicine including homeopathy for the treatment of glandular enlargements, cancerous tumours or hard lumps of testicles, prostate, ovaries, breasts and/ or uterus, particularly in the breast. However, if and how it acts still remains scientifically unknown. This study aims to test if Conium extract (CE), used as mother tincture of Conium in homeopathy, has demonstrable anti-cancer potentials without having much cytotoxicity in normal cells. Cytotoxicity of the drug was tested by conducting MTT assay on both normal (peripheral blood mononuclear cells) and HeLa cells. We also evaluated DNA fragmentation and DNA damage by DAPI and diphenylamine assay. The LDH activity assay was done to evaluate the percentages of apoptosis and necrosis. ROS accumulation also was evaluated to pin-point the actual events of apoptosis. Administration of drug clearly demonstrated its anti-cancer potentials as evidenced by the DNA damage analysis. The ROS activity also increased in case of the CE treated cells. LDH data revealed that the mode of cell death was mainly apoptotic and not necrotic. CE appears to induce apoptosis of cancer cells through ROS mediated pathway, and has negligible cytotoxicity against normal cells.