• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3395-3403
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• 2016

Conjugated linoleic acid, a functional lipid, produced from Lactobacillus plantarum (LP-CLA), has been demonstrated to possess apoptotic activity. The anti-proliferative and apoptotic potential of LP-CLA was here evaluated in vitro using the MDA-MB-231 human breast cancer cell line as a model system. Proliferation of MDA-MB-231 cells was inhibited with increasing concentrations of LP-CLA with altered morphological features like cell detachment, rounding of cells and oligonucleosomal fragmentation of DNA. Flow cytometry confirmed the apoptotic potential of LP-CLA by ANNEXIN V/PI double staining. Furthermore, outcome results indicated that the apoptosis was mediated by downregulation of the NF-${\kappa}B$ pathway which in turn acted through proteasome degradation of $I{\kappa}B{\alpha}$, inhibition of p65 nuclear translocation, release of cytochrome-C from mitochondria and finally overexpression of Bax protein. Thus, conjugated linoleic acid, a natural product derived from probiotics, could therefore be a possible potential chemotherapeutic agent due to its apoptotic activity against estrogen receptor negative breast cancer cells.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.895-901
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• 2013

Resistance to induction of apoptosis is a major obstacle for bladder cancer treatment. Bcl-2 is thought to be involved in anti-apoptotic signaling. In this study, we investigated the effect of Bcl-2 overexpression on apoptotic resistance and intracellular reactive oxygen species (ROS) generation in bladder cancer cells. A stable Bcl-2 overexpression cell line, BIU87-Bcl-2, was constructed from human bladder cancer cell line BIU87 by transfecting recombinant Bcl-2 [pcDNA3.1(+)-Bcl-2]. The sensitivity of transfected cells to adriamycin (ADR) was assessed by MTT assay. Apoptosis was examined by flow cytometry and acridine orange fluorescence staining. Intracellular ROS was determined using flow cytometry, and the activities of superoxide dismutase (SOD) and catalase (CAT) were also investigated by the xanthinoxidase and visible radiation methods using SOD and CAT detection kits. The susceptibility of BIU87-Bcl-2 cells to ADR treatment was significantly decreased as compared with control BIU87 cells. Enhanced expression of Bcl-2 inhibited intracellular ROS generation following ADR treatment. Moreover, the suppression of SOD and CAT activity induced by ADR treatment was blocked in the BIU87-Bcl-2 case but not in their parental cells. The overexpression of Bcl-2 renders human bladder cancer cells resistant to ADR-induced apoptosis and ROS might act as an important secondary messenger in this process.

• Journal of Oral Medicine and Pain
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• 제21권2호
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• pp.317-329
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• 1996

Allelic frequency and genotype distribution of short tandem repeat(STR) F13A01 locus was analysed by polymerase chain reaction, polyacrylamide gel electrophoresis and silver staining from human genomic deoxyribonucleic acid(DNA) was extracted from 205 unrelated Korean to be applied to forensic identification and parentage testing as a database. The results were as follows : 1. 5 alleles and 11 genotypes of F13A01 locus were detected and heterozygosity value was 62.0% and the observed each alleles and allelic frequency was 3.2(0.363), 4(0.105), 5(0.063), 6(0.466), 16(0.002). 2. The allelic diversity value was 0.639 and the power of discrimination was 0.804.3. Compared with observed number of alleles and allele frequency in ethnic difference, result was appeared to be similar to that of Japanese and Asians, while was appeared to be much different to that of Blacks and Caucasians in the observed number of alleles and frequency of allele 3.2, 5, 7. From the above result of this investigation, the allelic frequency of STR F13A01 locus in the Korean was considerd to be useful for individual identification and parentage testing as a database.

• Toxicological Research
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• 제27권2호
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• pp.71-76
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• 2011

We demonstrate that baicalein, a bioactive flavonoid originally isolated from Scutellaria baicalensis, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Treatment of peritoneal macrophages and RAW 264.7 cells with baicalein inhibited LPS-stimulated nitric oxide production in a dose-related manner. Immunohistochemical staining of iNOS and RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression in RAW 264.7 cells. Immunostaining of p65, EMSA, and reporter gene assay showed that baicalein inhibited NF-${\kappa}$B nuclear translocation, DNA binding, and transcriptional activation, respectively. Collectively, these series of experiments indicate that baicalein inhibits iNOS gene expression by blocking NF-${\kappa}$B nuclear translocation. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of baicalein on iNOS suggest that baicalein may represent a useful anti-inflammatory agent.

• Biomolecules & Therapeutics
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• 제4권4호
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• pp.402-407
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• 1996

Effects of oxidative stress on the induction of apoptosis and the activity of antioxidant enzymes were investigated in HL-60 cells using $H_2O$$_2$and cisplatin which generate oxygen species in the cell. Various concentrations of oxidants were treated to cells and at different incubation time, cells were harvested for assays. Cell viability, morphology by propidium iodide staining and DNA fragmentation by agarose gel electrophoresis were observed to determine whether they induce apoptosis. The activity of antioxidant enzymes such as superoxide dismutase and catalase was also measured to evaluate the cellular response to the oxidative damage. The results are as follows: $H_2O$$_2$ induced apoptosis at 10 $\mu$M after 6h incubation, while it took 12h for cisplatin. Both oxidants induced the superoxide dismutase activity at a tolerable low concentration. However, at a concentration which causes apoptotic cell death, the enzyme level was dropped markedly at first and then recovered to the normal level after which it declined again, probably due to cell death. On the other hand, changes in the activity of catalase were not significant at most concentrations except the statistically significant decrease at 24h after 10 $\mu$M-$H_2O$$_2$treatment. In this study, $H_2O$$_2$- and cisplatintreated cells showed similar results in apoptotic response and enzyme activities, suggesting that anticancer activity of cisplatin may be related, at least in part, to the production of oxygen free radicals.

• 한국식품영양학회지
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• 제20권2호
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• pp.108-113
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• 2007

In this study, a halophilic protease-producing bacterium was isolated from the west seaside mud flats of Korea. The 16S rDNA nucleotide sequences of the isolate showed 99.5% sequence homology with those of Vibrio vulnificus and Vibrio fluvialis; therefore, the isolate was named Vibrio sp. YH-127. Gram staining and the carbohydrate metabolism test results supported the isolate as one from the Vibrio family. Optimum condition for the cell growth and for the protease activity were obtained when the isolate was cultured at 25$^{\circ}C$ and pH 7.0, with the salt concentration of the medium similar to that of sea water. Finally, the addition of Mg$^{++}$ ions into medium increases protease activity suggesting that the protease produced by the isolate was a metalloprotease.

• Animal Systematics, Evolution and Diversity
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• 제31권4호
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• pp.277-283
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• 2015

Two pleurostomatid ciliates, Loxophyllum perihoplophorum Buddenbrock, 1920 and L. rostratum Cohn, 1866, were collected from the coastal waters of the East Sea, Korea. Their morphologies are described based on live observation and protargol staining, and morphometrics are provided. Loxophyllum perihoplophorum is characterized by the following features: 200-650 μm long in vivo; body slender leaf-shaped, flexible and contractile, with thin and wide extrusome-belted zone; 2 macronuclear nodules (Ma) and 1 micronucleus (Mi); 7-9 contractile vacuoles (CV) positioned along dorsal margin; extrusomes (Ex) evenly distributed along edge of entire body, with about 10 dorsal warts (Wa); 9-11 left (LSK) and 19-22 right somatic kineties (RSK), 4-5 furrows (Fu) on left side. Loxophyllum rostratum is about 100-130 μm long in vivo; body oblate leaf-shaped, contractile, convex ventral side and S-shaped dorsal side, beak-like anterior end; 2 Ma and 1 Mi; 1 CV terminally located; Ex distributed along edge of entire body, with about 9-10 dorsal Wa; 7-8 LSK and 15-19 RSK, ca. 5 Fu on left body side. In addition, sequences of small subunit ribosomal DNA were determined from these two Loxophyllum species and compared with the known Loxophyllum sequences.

• 동의생리병리학회지
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• 제19권4호
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• pp.955-959
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• 2005

We previously demonstrated that the methylene chloride fraction of Scutellaria barbata suppessed human leukemic U937 cell proliferation by inducing apoptosis. In the present study, we have isolated luteolin from Scutellaria barbata and evaluated its apoptotic mechanism in Lewis lung carcinoma cells. Luteolin inhibited the proliferation of Lewis lung carcinoma cells in a concentration-dependent manner. Luteolin effectively increased the portion of $sub-G_1$ DNA content (apoptotic portion) and apoptotic Annexin-V positive cells in a concentration-dependent manner by FACS analysis. Caspase 9 and caspase 3 were activated and PARP was effectively cleaved by luteolin. It also increased the ratio of Bax to Bcl-2 through the decrease of Bcl-2 expression by Western blotting and reduced mitochondrial membrane potential following TMRE staining. These results suggest that luteolin can induce apoptosis through the mitochondrial mediated pathway.

• Food Science and Biotechnology
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• 제16권4호
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• pp.537-542
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• 2007

Sophora flavescens AITON (Leguminosae) is a typical traditional Korean medical herb considered to exhibit antibacterial, anti-inflammatory, and antipyretic effects, and is also used for the treatment of skin and mucosal ulcers, sores, diarrhea, gastrointestinal hemorrhage, arrhythmia, and eczema. In this study, the compound sophoraflavanone G was isolated from the dried roots of S. flavescens by bioassay-guided fractionation. We then investigated the effects of various concentrations of sophoraflavanone G on cell viability and the induction of apoptosis in KB cells after an incubation of 24 hr. The results were determined by the following methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-terazolium bromide (MTT) assay, Hoechst-33258 dye staining, flow cytometry (cell cycle), and Western blotting for caspase-3 and poly (ADP-ribose) polymerase (PARP). We found sophoraflavanone G induced the apoptosis of KB cells in a dose-dependent manner that was verified by DNA fragmentation, apoptotic bodies, the sub-G1 ratio, caspase-3 activity, and cleavage of PARP. These results suggest that sophoraflavanone G has potent anti-proliferative effects on human oral epidermoid carcinoma cells, with the induction of apoptosis.

• Korean Journal of Acupuncture
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• 제20권1호
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• pp.57-64
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• 2003

목적: 오가피가 mouse testis에서 유래된 Leydig 세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향을 조사하였다. 방법: TM3 세포주에서의 아폽토시스 변화를 관찰하기 위해서 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI), DNA fragmentation assay, 및 reverse transcription-polymerase chain reaction (RT-PCR) 방법을 이용하였다. 결과: MTT assay를 이용하여 분석한 결과 농도에 따른 세포독성의 효과가 에탄올 투여부터 관찰되었다. 또한 오가피로 전처치하고 에탄올을 처치하였을 때 세포독성이 크게 감소되었다. DAPI staining에서 오가피 투여군은 에탄올 투여군에 비해서 fragmentation이 억제되었다. RT-PCR의 분석에 의하여 caspase-3 mRNA 발현이 오가피 투여군은 알코올 투여군보다 유의성있게 억제됨을 보여주었다. 결론: TM3 Leydig 세포주에서 에탄올에 의해 유발된 아폽토시스는 전형적인 세포사멸 형태를 나타내었다. 반면에 오가피 투여군은 에탄올에 의해서 유발된 아폽토시스에서 세포보호 효과가 있음이 확인되었다.