• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4331-4338
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• 2014

$N^1$-(2, 5-dimethoxyphenyl)-$N^8$-hydroxyoctanediamide (N25) is a novel SAHA cap derivative of HDACi, with a patent (No. CN 103159646). This invention is a hydroxamic acid compound with a structural formula of $RNHCO(CH_2)6CONHOH$ (wherein R=2, 5dimethoxyaniline), a pharmaceutically acceptable salt which is soluble. In the present study, we investigated the effects of N25 with regard to drug distribution and molecular docking, and anti-proliferation, apoptosis, cell cycling, and $LD_{50}$. First, we designed a molecular approach for modeling selected SAHA derivatives based on available structural information regarding human HDAC8 in complex with SAHA (PDB code 1T69). N25 was found to be stabilized by direct interaction with the HDAC8. Anti-proliferative activity was observed in human glioma U251, U87, T98G cells and human lung cancer H460, A549, H1299 cells at moderate concentrations ($0.5-30{\mu}M$). Compared with SAHA, N25 displayed an increased antitumor activity in U251 and H460 cells. We further analyzed cell death mechanisms activated by N25 in U251 and H460 cells. N25 significantly increased acetylation of Histone 3 and inhibited HDAC4. On RT-PCR analysis, N25 increased the mRNA levels of p21, however, decreased the levels of p53. These resulted in promotion of apoptosis, inducing G0/G1 arrest in U251 cells and G2/M arrest in H460 cells in a time-dependent and dose-dependent manner. In addition, N25 was able to distribute to brain tissue through the blood-brain barrier of mice ($LD_{50}$: 240.840mg/kg). In conclusion, our findings demonstrate that N25 will provide an invaluable tool to investigate the molecular mechanism with potential chemotherapeutic value in several malignancies, especially human glioma.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.9
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• pp.1143-1150
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• 2017

The aim of this study was to determine the chemical and intracellular antioxidant activities of ${\beta}$-carotene and lycopene and to compare their quantitative structure-activity relationship (QSAR). In our previous study, the second ionization energy of lycopene was higher than that of ${\beta}$-carotene, as calculated by QSAR. Chemical antioxidant activities of ${\beta}$-carotene, lycopene, and Trolox were examined by measuring ferric reducing antioxidant power (FRAP) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Intracellular antioxidant activities were evaluated by intracellular reactive oxygen species (ROS) and DNA fragmentation. The FRAP of lycopene was higher than that of ${\beta}$-carotene (P<0.05), and the two carotenoids had similar antioxidant activities in DPPH radical scavenging activity assay. Trolox had the greatest chemical antioxidant activities (P<0.05). When RAW264.7 cells were treated with lipopolysaccharide (LPS) (100 ng/mL) for 20 h, intracellular ROS and DNA fragmentation significantly increased (P<0.05). RAW 264.7 cells pretreated with ${\beta}$-carotene ($4{\mu}M$) and lycopene ($0.4{\sim}2{\mu}M$) for 4 h formed significantly less intracellular ROS than LPS-treated control cells (P<0.05), whereas cells with Trolox did not reduce production of intracellular ROS. In addition, cells pretreated with $2{\mu}M$ lycopene produced less intracellular ROS than those treated with ${\beta}$-carotene (P<0.05). DNA fragmentation of cells with ${\beta}$-carotene and lycopene was similar to that of LPS-treated control cells as measured by Hoechst staining. The antioxidant ability of lycopene was greater than that of ${\beta}$-carotene in the QSAR, FRAP, and intracellular ROS assays (P<0.05). ${\beta}$-Carotene and lycopene had lower antioxidant activities as measured by FRAP (P<0.05) but higher intracellular protective effects against LPS-induced oxidative stress in comparison with Trolox.

• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.511-518
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• 1998

Anti-complementary assay for immuno-stimulating polysaccharide screening, human tumor colony-forming assay for discovering anti-tumor drugs, and toxic assay against mouse were performed to examine pharmacological activities of polysaccharides extracted from fruiting bodies of Jang-soo mushroom (Fomitella fraxinea). Hot water $(100^{\circ}C,\;FF-I)$, 1% ammonium oxalate solution $(80^{\circ}C,\;FF-II)$, and 5% sodium hydroxide solution $(80^{\circ}C,\;FF-III)$ were used for extraction of three polysaccharides from fruiting bodies of it. Anti-complementary activity of FF-I was more effective than the others. FF-I was further fractionated into three groups of polysaccharide by DEAE-Sephadex A25 column chromatography (FF-NP, FF-AP1, and FF-AP2). FF-AP1 showed not only the highest anti-complementary activity but also the growth-inhibitory activity against Snu-I (human stomach cancer cell) among 9 kinds of human tumor cell lines. But FF-AP2 exhibited its activity against Hep-2(larynx cancer) and KB(mouth epidermal cancer) cell lines at $500\;{\mu}g/ml$ although its anti-complementary activity was lower than it of FF-AP1. When FF-I was orally administrated to mice with dosage of 5000 mg/kg, no remarkable changes were observed in viewpoint of tissue-pathology.

• Journal of Photoscience
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• v.9 no.2
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• pp.182-185
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• 2002

There are 3 UV DNA repair endonuclease activities in mammalian cells that cleave UV -irradiated DNA. Interestingly, mammalian UV endonuclease III with MW of 26.7kD has a lyase activity on AP sites. It also cleaves the phosphodiester bond within a cyclobutane pyrimidine dimer. Genomic analysis of human repair endonuclease III gene revealed that this gene has 100% sequence identity with ribosomal protein S3 (rpS3). Therefore, rpS3 seems to function both in translation and in DNA repair. This gene of about 6.1 kb contains 6 introns and 7 exons, and the first and fifth introns of human rpS3 gene contain functional U15 small nucleolar (sno) RNAs which appear to be involved in ribosome assembly. It is to be noted that the column profile of the endonuclease activity of rpS3 appears to be altered in Xeroderma Pigmentosum (XP) group D cells compared to normal cells indicating that this protein is involved in XP disease as well. XP is a human disease characterized by high sensitivity of skin by UV- or sun-light irradiation and by high frequency of developing skin cancers. We also report here that rpS3 protein is involved in other cellular functions.

• Toxicological Research
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• v.16 no.2
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• pp.95-100
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• 2000

In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

• Biomolecules & Therapeutics
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• v.24 no.4
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• pp.380-386
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• 2016

Silymarin from milk thistle (Silybum marianum) has been reported to show an anti-cancer activity. In previous study, we reported that silymarin induces cyclin D1 proteasomal degradation through NF-${\kappa}B$-mediated threonine-286 phosphorylation. However, mechanism for the inhibition of Wnt signaling by silymarin still remains unanswered. Thus, we investigated whether silymarin affects Wnt signaling in human colorectal cancer cells to elucidate the additional anti-cancer mechanism of silymarin. Transient transfection with a TOP and FOP FLASH luciferase construct indicated that silymarin suppressed the transcriptional activity of ${\beta}$-catenin/TCF. Silymarin treatment resulted in a decrease of intracellular ${\beta}$-catenin protein but not mRNA. The inhibition of proteasome by MG132 and $GSK3{\beta}$ inhibition by SB216763 blocked silymarin-mediated downregulation of ${\beta}$-catenin. In addition, silymarin increased phosphorylation of ${\beta}$-catenin and a point mutation of S33Y attenuated silymarin-mediated ${\beta}$-catenin downregulation. In addition, silymarin decreased TCF4 and increased Axin expression in both protein and mRNA level. From these results, we suggest that silymarin-mediated downregulation of ${\beta}$-catenin and TCF4 may result in the inhibition of Wnt signaling in human colorectal cancer cells.

• Korean Journal of Plant Resources
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• v.22 no.4
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• pp.312-316
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• 2009

In the present study, the anti-cancer activity of 80% ethanol extracts from 120 kinds of medicinal herbs and native plants were investigated. Among them, the barks of Melia azedarach L. var. japonica Makino showed the highest cytotoxicity in HCT-15 human colon cancer cell. With this result, we carried out hollow fiber (HF) assay and anti-metastasis study to confirm the anti-cancer effects of M. azedarach var. japonica. In MTT assay, M. azedarach var. japonica.inhibited the proliferation of HCT-15 cells in dose-dependent manner. HF assay was carried out using A549 human adenocarcinoma cell, HCT-15 and SK-Hep1 human liver cancer cell via intraperitoneal (IP) and subcutaneous (SC) site. As a results, SK-Hep1 implanted in IP site showed the highest cytotoxicity. The result from metastatic model using B16/BL6 mouse corresponded to that of HF assay. These results suggest that the ethanol extract from M. azedarach var. japonica. might have a potent anti-cancer activity and advanced study is needed for the development of novel natural anti-cancer drug.

• The Journal of Korean Medicine
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• v.27 no.4
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• pp.162-173
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• 2006

The water-extracts of Scutellaria barbata Don (SBDE) were isolated from Chinese medicinal plant sources. The extracts showed strong growth-inhibitory activity and cancer chemopreventive activity on the growth and DNA incorporation of MG63 human osteosarcoma and K562 human leukemia cell lines. The growth of human cancer cells was inhibited in the presence of the extracts (20, 50 and 100 ${\mu}$g/ml), and the effects were concentration-dependent and incubation time-dependent up to 8 days. When 50 ${\mu}$g/ml of the extracts was added to the media of MG63 and K562, cell growth after 8 days or 6 days of incubation was retarded by 93.2 to 97.3% of the control group. Morphological changes of MG63 and K562 cell lines were observed. As the concentration of the extracts increased up to 50 ${\mu}$g/ml, degree of cell aggregation decreased. Moreover, the DNA incorporation of the cells which were labeled with [3H] thymidine was significantly reduced after 3 days of incubation at $37^{\circ}C$ with the extract. Therefore, it is suggested that the extract is highly effective on inhibition of cancer cell growth. The extract also inhibited gene expression of IGF-II in transcriptional level. Since IGF-II works as a mitogenic effector on MG63 and K562 cell lines, these results suggest that the growth inhibition is in part mediated through the inhibition of IGF-II gene expression.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1139-1141
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• 2009

Clitocybin D, a novel human neutrophil elastase inhibitor, was isolated from the culture broth of Clitocybe aurantiaca. This compound was purified by solvent extraction, silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative HPLC. The compound was determined to be 4-(4,6-dihydroxy-3-methoxy-3H-isoindol-1-yl)-benzoic acid on the basis of 1D and 2D NMRs and MS spectroscopic analysis. Analysis of the human neutrophil elastase (HNE) inhibitory activity of the isolated compound revealed that it showed significant HNE inhibitory activity with an $IC_{50}$ value of $17.8{\mu}M$.

• Nutrition Research and Practice
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• v.3 no.2
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• pp.77-83
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• 2009

Retinoic acids (RAs) modulate growth, differentiation, and apoptosis in normal, pre-malignant & malignant cells. In the present study, the effects of RA isomers (all-trans RA, 13-cis RA, and 9-cis RA) on the cell signal transduction of human breast cancer cells have been studied. The relationship between RAs and an enzymatic antioxidant system was also determined. Estrogen-receptor (ER) positive MCF-7 and ER-negative MDA-MB-231 human breast cancer cells were treated with different doses of each RA isomers, all-trans RA, 13-cis RA, or 9-cis RA. Treatment of RA isomers inhibited cell viability and induced apoptosis of MCF-7 cells as a result of increased caspase activity in cytoplasm and cytochrome C released from mitochondria. All-trans RA was the most effective RA isomer in both cell growth inhibition and induction of apoptosis in MCF-7 cells. However, no significant effect of RA isomers was observed on the cell growth or apoptosis in ER-negative MDA-MB-231 cells. In addition, activities of antioxidant enzymes such as catalase and glutathione peroxidase were decreased effectively after treatment of RA in MCF-7 cells, whereas SOD activity was rarely affected. Thus, the present data suggest that all-trans RA is the most potential inducer of apoptosis and modulator of antioxidant enzymes among RA isomers in MCF-7 human breast cancer cells.