• 대한한방내과학회지
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• 제25권2호
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• pp.202-213
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• 2004

Object : Objective: This study was conducted to investigate the anti-cancer effects of Mylabris phalerata (반모) in some kinds of cancer cells. Materials and Methods: Some kinds of cancer cells lines were treated. We used nine kinds of cancer cell lines, such as stomach cancer cells (Kato), lung cancer cells (Calu-1, NCI-H 1395), urinary bladder cancer cells (HS789T), bone cancer cells (Saos-2), brain cancer cells (SK-N-MC), liver cancer cells (Hep-G2), skin cancer cells (Mo-1) and prostate cancer cells (PC-3) with the water decoction of Mylabris phalerata. The histological changes of all cell lines in the media (RPMI-1640) containing the decoction of Mylabris phalerata were observed and we examined cell death assay by trypan blue exclusion testing was examined. Finally, the change of mitochondrial membrane potential was measurd and the inhibitory effect of Mylabris phalerata on cell increase was examined by analyzing the cell cycle. Results: In histologic change all cancer cell lines showed withdrawn and floating appearance that is typical in cellular impairment. Most of the cell lines showed over 50% death rate after 24 hours in trypan blue exclusion tests. Especially the stomach, urinary bladder. brain and liver cell lines showed over 30% death rate after 12 hours. All cell lines treated with Mylabris phalerata were less stained than the control group and the mitochondrial membrane potential in the Mylabris phalerata treated cell lines was markedly lower than that in the control group. The measurement of DNA quantity in all cell lines showed the disappearance of the peak and the thickened left axis, which suggests that all cellular DNA degraded. Conclusion: Mylabris phalerata had cytotoxicity on various kinds of cancer cell lines and the mechanism of that was the impairment of mitochondria by the breakdown of the mitochondrial cell membrane. We propose that this is in part attributable to the destruction of DNA in cancer cells.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.435-442
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• 2001

Poly(3-hydroxy-5-phenylvalerate) [P(3HPV)] was efficiently accumulated from 5-phenylvalerate (5PV) in Pseudomonas putida BM01 in a mineral salts medium containing butyric acid (BA) as the cosubstrate. A nove aromatic copolyester, poly(5 mol% 3-hydroxy-4-phenylbutyrate-co- 95 mol% 3-hydroxy-6-phenylhexanoate) [P(3HPB-co-3HPC)] was also synthesized from 6-phenylhexanoate (6PC) plus Ba. The two aromatic polymers, P(3HPV) and P(3HPB-co-3HPC), were found to be amorphous and showed different glass-transition temperatures at $15^{\circ}C$ and $10^{\circ}C$, respectively. When the bacterium was grown ina medium containing 20 mM 5PV as the sole carbon source for 140 h, 0.4 g/l of dry cells was obtained in a flask cultivation and 20 wt% of P(3HPV) homopolymer was accumulated in the cells. However, when it was grown with a mixture of 2 mM 5PV and 50 mM BA for 40 h, the yield of dry biomass was increased up to 2.5 g/l and the content of P(3HPV) in the dry cells was optimally 56 wt%. This efficient production of P(3HPV) homopolymer from the mixed substrate was feasible because BA only supported cell growth and did not induce any aliphatic PHA accumulation. The metabolites released into the PHA synthesis medium were analyzed using GC or GC/MS. Two $\beta$-oxidation derivatives, 3-phenylpropionic acid and trans-cinnamic acid, were found in the 5V-grown cell medium and these comprised 55-88 mol% of the 5PV consumed. In the 6PC-grown medium containing Ba, seven ${\beta}$-oxidation and related intermediates were found, which included phenylacetic acid, 4-phenylbutyric acid, cis-4-phenyl-2-butenoic acid, trans-4-phenyl-3-butenoic acid, trans-4-phenyl-2-butenoic acid, 3-hydroxy-4-phenylbutyric acid, and 3-hydroxy-6-phenylhexanoic acid. Accordingly, based on the metabolite analysis, PHA synthesis pathways from the two aromatic carbon sources are suggested.

• 대한약침학회지
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• 제18권3호
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• pp.32-41
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• 2015

Objectives: Condurango (Gonolobus condurango) extract is used by complementary and alternative medicine (CAM) practitioners as a traditional medicine, including homeopathy, mainly for the treatment of syphilis. Condurango bark extract is also known to reduce tumor volume, but the underlying molecular mechanisms still remain unclear. Methods: Using a cervical cancer cell line (HeLa) as our model, the molecular events behind condurango extract's (CE's) anticancer effect were investigated by using flow cytometry, immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Other included cell types were prostate cancer cells (PC3), transformed liver cells (WRL-68), and peripheral blood mononuclear cells (PBMCs). Results: Condurango extract (CE) was found to be cytotoxic against target cells, and this was significantly deactivated in the presence of N-acetyl cysteine (NAC), a scavenger of reactive oxygen species (ROS), suggesting that its action could be mediated through ROS generation. CE caused an increase in the HeLa cell population containing deoxyribonucleic acid (DNA) damage at the G zero/Growth 1 (G0/G1) stage. Further, CE increased the tumor necrosis factor alpha ($TNF-{\alpha}$) and the fas receptor (FasR) levels both at the ribonucleic acid (RNA) and the protein levels, indicating that CE might have a cytotoxic mechanism of action. CE also triggered a sharp decrease in the expression of nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) both at the RNA and the protein levels, a possible route to attenuation of B-cell lymphoma 2 (Bcl-2), and caused an opening of the mitochondrial membrane's permeability transition (MPT) pores, thus enhancing caspase activities. Conclusion: Overall, our results suggest possible pathways for CE mediated cytotoxicity in model cancer cells.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of International Convention of the Pharmaceutical Society of Korea
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• pp.109-113
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• 2001

Oxidized low-density lipoprotein (oxLDL) has been shown to modulate transactivations by the peroxisome proliferator activated receptor (PPAR)$\gamma$ and nuclear factor-kappa B (NF$\kappa$B). In this study, the oxLDL signaling pathways involved with the NF$\kappa$B transactivation were investigated by utilizing a reporter construct driven by three upstream NF$\kappa$B binding sites, and various pharmacological inhibitors. OxLDL and its constituent lysophophatidylcholine (lysoPC) induced a rapid and transient increase of intracellular calcium and stimulated the NF-KB transactivation in resting RAW264.7 macrophage cells in an oxidation-dependent manner. The NF$\kappa$B activation by oxLDL or lysoPC was inhibited by protein kinase C inhibitors or an intracellular calcium chelator. Tyrosine kinase or PI3 kinase inhibitors did not block the NF$\kappa$B transactivation. Furthermore, the oxLDL-induced NF$\kappa$B activity was abolished by the PPAR$\gamma$ ligands. When the endocytosis of oxLDL was blocked by cytochalasin B, the NF$\kappa$B transactivation by oxLDL was synergistically increased, while PPAR transactivation was blocked. These results suggest that oxLDL activates NF-$\kappa$B in resting macrophages via protein kinase C- and/or calcium-dependent pathways, which does not involve the endocytic processing of oxLDL. The endocytosis-dependent PPAR$\gamma$ activation by oxLDL may function as an inactivation route of the oxLDL induced NF$\kappa$B signal. Short heterodimer partner (SHP), specifically expressed in liver and a limited number of other tissues, is an unusual orphan nuclear receptor that lacks the conventional DNA-binding domain. In this work, we found that SHP expression is abundant in murine macrophage cell line RAW 264.7 but suppressed by oxLDL and its constituent I3-HODE, a ligand for peroxisome proliferator-activated receptor y. Furthermore, SHP acted as a transcription coactivator of nuclear factor-$\kappa$B (NF$\kappa$B) and was essential for the previously described NF$\kappa$B transactivation by lysoPC, one of the oxLDL constituents. Accordingly, NF$\kappa$B, transcriptionally active in the beginning, became progressively inert in oxLDL-treated RAW 264.7 cells, as oxLDL decreased the SHP expression. Thus, SHP appears to be an important modulatory component to regulate the transcriptional activities of NF$\kappa$B in oxLDL-treated, resting macrophage cells.

• 농업생명과학연구
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• 제45권4호
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• pp.113-120
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• 2011

본 연구에서는 생마늘 추출 숙성물의 용매 분획물에 대한 항산화 효과와 acetylcholinesterase (AChE) 저해 효과에 대해서 조사하였다. 생마늘 추출 숙성물의 헥산, 클로포름, 에틸아세테이트 및 물 분획물을 통한 총 페놀성 화합물의 함량은 각각 3.70, 23.63, 31.27, 2.35 mg/g으로 나타났다. 또한, 에틸아세테이트 분획물이 ABTS radical scavenging activity, ferric reducing antioxidant power 및 linoleic acid를 활용한 지질의 자동산화 저해 효과에서 가장 높다는 것을 확인하였다. 추가적으로, PC12 신경 세포에서의 에틸아세테이트 분획물의 처리가 $H_2O_2$가 유발시키는 산화적 스트레스의 수준을 감소시켰다. 결국 생마늘 추출 숙성물의 에틸아세테이트 분획물이 농도 의존적으로 AChE를 저해한다는 것을 확인하였다. 그러므로 본 연구는 생마늘 추출 숙성물의 에틸아세테이트 분획물이 PC12 신경세포에 있어서 산화적 스트레스 수준을 감소시키고, 또한 AChE 저해제로서 활용될 수 있는 가능성을 확인하였다.

• 한국자원식물학회지
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• 제36권3호
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• pp.237-255
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• 2023

Variations in phenotypic traits are important for onion genetic improvement. The aim of this study was to identify the phenotypic traits of temporary genetic resources and the best accessions for the development of onion breeding programs. Sixteen phenotypic traits of 79 onion accessions were studied. The descriptive statistics of phenotypic traits exhibited a high variation in onion accessions. Among the 79 evaluated accessions, 64.55% had a large bulb neck width and 44.30% had a circular bulb shape. Principal component analysis showed that six principal components (PCs) accounted for 72.65% of the total variation. The main factors contributing to PC1 were bulb weight, equatorial and bulb polar diameters, plant height, and degree of splitting into bulblets, whereas those contributing to PC2 were the bulb color of the epidermal cells of the fleshy scales and color of the dry skin on the bulb. The accessions were classified into three groups-clusters 1, 2, and 3. Cluster 2 accessions were the most suitable for selecting large and circular bulb-shaped onion cultivars. The phenotypic variation observed in this study may help to select potential accessions for breeding new onion cultivars.

• 생명과학회지
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• 제18권9호
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• pp.1312-1315
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• 2008

DAPI 염색을 이용하여 세포사멸 유도물질의 초고속스크리닝에 적합한 protocol을 제작하였다. Liquid handler에서 dispensing과 multilabel counter에서의 상대적인 fluorescent intensity를 측정하는 과정을 자동화한 protocol을 제작하였고, 이의 과정은 매우 효율적으로 가동되었음을 확인하였다. DAPI에 binding된 DNA의 intensity를 측정하는 조건검토에서 가장자리의 36개를 reading하지 않는 것이 edge effect를 줄일수 있는 것을 확인하였고, 0.1 sec reading으로 시간을 절약할 수 있었다. 이의 결과를 효과적으로 이용할 경우, 세포사멸 유도물질의 초고속스크리닝 또는 이에 적합한 스크리닝이 가능함을 확인하였다.

• 대한의생명과학회지
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• 제9권2호
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• pp.67-74
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• 2003

Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

• Molecules and Cells
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• 제38권2호
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• pp.171-179
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• 2015

Prostatic acid phosphatase (PAP) expression increases proportionally with prostate cancer progression, making it useful in prognosticating intermediate to high-risk prostate cancers. A novel ligand that can specifically bind to PAP would be very helpful for guiding prostate cancer therapy. RNA aptamers bind to target molecules with high specificity and have key advantages such as low immunogenicity and easy synthesis. Here, human PAP-specific aptamers were screened from a 2'-fluoropyrimidine (FY)-modified RNA library by SELEX. The candidate aptamer families were identified within six rounds followed by analysis of their sequences and PAP-specific binding. A gel shift assay was used to identify PAP binding aptamers and the 6N aptamer specifically bound to PAP with a Kd value of 118 nM. RT-PCR and fluorescence labeling analyses revealed that the 6N aptamer bound to PAP-positive mammalian cells, such as PC-3 and LNCaP. IMR-90 negative control cells did not bind the 6N aptamer. Systematic minimization analyses revealed that 50 nucleotide sequences and their two hairpin structures in the 6N 2'-FY RNA aptamer were equally important for PAP binding. Renewed interest in PAP combined with the versatility of RNA aptamers, including conjugation of anti-cancer drugs and nano-imaging probes, could open up a new route for early theragnosis of prostate cancer.

• Journal of Acupuncture Research
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• 제22권5호
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• pp.37-48
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• 2005

Objectives : The purpose of this study was to investigate the anti-caner effect of cobrotoxin on the prostatic cancer cell line(PC-3). Methods : After the treatment of PC-3 cells with cobrotoxin, we performed fluorescence microscope, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify $NF-{\kappa}B$ the change of calcium and NO. Results : 1. The expression of $NF-{\kappa}B$ was decreased at 1nM and It·as decreased significantly at 2, 4, 8nM. 2. $I{\kappa}B,\;NF-{\kappa}B$ inhibitor, was decreased significantly at 8nM and $p-l{\kappa}Ba$, phosphrylation of $I{\kappa}B$, was decreased significantly at all concentrations of cobrotoxin. 3. The expressions of p50 and p65 were decreased significantly and dose-dependently at 1, 2, 4, 8nM. 4. The expression of p53 was increased significantly at 1, 2, 4, 8nM. 5. The calcium concentration in cell wasn't changed at 1, 2, 4, 8nM, but was increased dose-dependently at 30, 70, 130, 250nM comparing with lower dose of cobrotoxin. 6. The NO concentration in cell was increased significantly at 1, 2, 4, 8nM. 7. In immunochemical staining, we found that cobrotoxin-immunochemical complex move into intracellular space dose-dependently. Conclusion : These results indicate that cobrotoxin has anti-cancer effects by inducing apoptosis.