• Journal of Ginseng Research
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• v.45 no.1
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• pp.48-57
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• 2021

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

• Korean Journal of Medicinal Crop Science
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• v.12 no.1
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• pp.36-42
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• 2004

The immune activation and anticancer activities of the water and ethanol extracts from Artemisia capillaris Thunb. were studied. The growth of human hepatocarcinoma and human gastric cancer cell was inhibited by the addition of $1.0\;mg/m{\ell}$ of the water extract, by about 77% and 95%, respectively. The growth of human breast cancer cells was also inhibited by addition of $0.5\;mg/m{\ell}$ of both water and ethanol extracts by 88%. The growth of human normal lung cell, HEL299 was inhibited by 15% indicating very low cytotoxicity of both extracts. Overall selectivity of the both extracts on several human cancer cell line was over 2.5. The growth of both human B and T cells was enhanced up to 1.6 to 2.1 times by adding the ethanol extracts. The secretion of cytokines, $TNF-{\alpha}$ and IL-6, from human B cells was also increased showing $68\;pg/m{\ell}$ and $67\;pg/m{\ell}$, respectively, compared to $35{\sim}40\;pg/m{\ell}$ of the control. In terms of the immune activity, there was not much difference between water and ethanol extracts of Artemisia capillaris Thunb. It implies that the extraction solvent could not differ the biological activities of the extracts. Based on these results, Artemisia capillaris Thunb. can be developed into a potentially useful cancer chemoprentive agent.

• Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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• v.1 no.1
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• pp.53-90
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• 2005

Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.20
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• pp.8693-8698
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• 2014

Background: Up-regulation of hsp90 gene expression occurs in numerous cancers such as lung cancer. D,L-lactic-co-glycolic acid-poly ethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG may inhibit the expression. The purpose of this study was to examine whether nanocapsulating 17DMAG improves the anti cancer effect over free 17DMAG in the A549 lung cancer cell line. Materials and Methods: Cells were grown in RPMI 1640 supplemented with 10% FBS. Capsulation of 17DMAG is conducted through double emulsion, then the amount of loaded drug was calculated. Other properties of this copolymer were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity on the grown of lung cancer cell line was carried out through MTT assay. After treatment, RNA was extracted and cDNA was synthesized. In order to assess the amount of hsp90 gene expression, real-time PCR was performed. Results: In regard to the amount of the drug load, IC50 was significant decreased in nanocapsulated(NC) 17DMAG in comparison with free 17DMAG. This was confirmed through decrease of HSP90 gene expression by real-time PCR. Conclusions: The results demonstrated that PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of hsp90 expression by enhancing uptake by cells. Therefore, PLGA-PEG could be a superior carrier for this kind of hydrophobic agent.

• Korean Journal of Plant Resources
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• v.24 no.1
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• pp.40-47
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• 2011

Sphagnum palustre is a semi aquatic moss. S. palustre has been used as Korean traditional medicine to treat cardiac pain and stroke. This study was carried out to analyze phytochemical constituents of S. palustre and investigate the biological activity for the promotion of human health. At first, we isolated seven compounds from the ethanolic extract of this plant, and their structures were characterized by spectroscopic methods. Their structures were characterized to be Coumarin(1), Caffeic acid(2), Quercetin(3), Astragalin(4), Luteolin(5), Chlorogenic acid(6), Rutin(7) were for the first time reported from this source. The ethanol extract from S. palustre which was tested for its anticancer activity against three human tumor cell line by in vitro assay.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.57 no.4
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• pp.353-358
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• 2012

This study was carried out to evaluate antioxidant and anticancer activity of crudes extracts from colored corn (Zea mays. L.) among particle size with different pulverizing methods (pin mill and ultra fine pulverizer). In cytotoxicity test using extracts from the flours grounded by pin mill and ultra fine pulverizer respectively, it showed that A-549 was the highest anticancer activity among in vitro using 4 cancer cell line types (Hep-G2, A-549, HCT-116 and MCF-7). Assessing the antioxidant activities of crude extracted from different pulverizing methods was measured by trolox equivalent antioxidant capacity (TEAC) and ferric reducing antioxidant power (FRAP). Higher scanvening activity against free radical was observed in the crude extracted from small particle size flours of colored corn than in those of the big particle size.

• Food Science and Preservation
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• v.20 no.5
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• pp.691-698
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• 2013

The biological activities of Smilax china L. rhizome (SCR), hot water (SCRW) and 70% ethanol extract (SCRE) were analyzed. The total phenolic contents of SCRW and SCRE were 51.7 and 100.5 mg/g, respectively. The measured flavonoid content of SCRW ($67.7{\mu}g/g$) was almost double that of SCRE ($31.7{\mu}g/g$). SCRE ($IC_{50}=42.4{\mu}g/mL$) exhibited stronger antioxidant activity in the DPPH system than the positive control ${\alpha}$-tocopherol ($71.3{\mu}g/mL$) or butylated hydroxy anisole ($53.8{\mu}g/mL$) did. SCRE ($IC_{50}=50.3{\mu}g/mL$) also showed stronger ABTS radical scavenging activity, as did ${\alpha}$-tocopherol ($67.1{\mu}g/mL$). The SOD-like activity and Tyrosinase inhibition activity of SCRW and SCRE showed almost the same pattern. The best SOD-like activity and tyrosinase inhibition activity were measured as 24.9% and 20.3% in SCRW at $1,000{\mu}g/mL$, respectively. The cytotoxic effects of the SCR extracts were analyzed via MTT assay on human cancer and normal cells. SCRW and SCRE did not show cytotoxicity up to the concentration of $1,000{\mu}g/mL$ against the normal human cell line HEK293. Against human breast cancer cells (MCF-7), SCRW inhibited MCF-7 growth (by 27.6%) better than the anticancer drug cyclophosphamide (15.5%) at $1,000{\mu}g/mL$. SCRE ($1,000{\mu}g/mL$) inhibited the growth of human lung cancer cells A549 (37.6%) and human stomach cancer cells AGS (53.6%) more effective than did SCRW (21.0% and 35.4%) or CPA (22.2% and 31.7%). These results suggest the potential use of SCRE and SCRW as an excellent antioxidant and antiproliferative substance, respectively.

• Food Science and Preservation
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• v.14 no.2
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• pp.220-225
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• 2007

This study was performed to measure the antioxidative, antimutagenic, and cytotoxic properties of Prunus armeniaca using the DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical donating method, the Ames test, and cytotoxicity measurements, respectively. Electron-donating abilities were 48.3, 43.9, 14.8 and 12.9 per g dry matter of P. armeniaca seed (PAS), P. armeniaca flesh(PAF), butylated hydroxytoluene, and ${\alpha}-tocopherol$, respectively. The direct antimutagenic effects of an ethanol extract of P. armeniaca were examined in Ames tests using Salmonella typhimurium TA98 and TA100 as reporter organisms. In the Ames test, the ethanol extract of P. armenicaca alone did not exhibit any mutagenicity but the extract did show substantial inhibitory effects against mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitroquinoline-1-oxide(4NQO). The ethanol extract of PAS(200g dry matter/plate) inhibited strain TA98 mutagenesis induced by 4NQO by ca. 37.9%, and mutation inhibition values of 42.1% and 69.4%, respectively, were observed when 4NQO and MNNG acted on the TA100 strain. The cytotoxic effects of ethanol extracts of P. armeniaca against cell lines of human lung carcinoma(A549), human breast adenocarcinoma(MCF-7), human hepatocellular carcinoma(Hep3B), human cervical adenocarcinoma(HeLa), and human gastric carcinoma(AGS) rose with increases in extract concentration. An ethanol extract(4mg/mL dry matter) of PAF showed strong cytotoxicities of 88.2%, 58%, 72.8%, 89.4%, and 91.9% against A549, AGS, MCF-7, HeLa, and Hep3B cells, respectively. In contrast, the same extract showed only 13 37% cytotoxicity for a nomal human kiney cell line(293). It is suggested that P. armeniaca possesses useful antioxidative, antimutagenic, and anticancer properties.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.69-73
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• 2014

The effects of erlotinib combined with celecoxib in a lung cancer xenograft model were here explored with a focus on possible mechanisms. A xenotransplanted lung cancer model was established in nude mice using the human lung cancer cell A549 cell line and animals demonstrating tumour growth were randomly divided into four groups: control, erlotinib, celecoxib and combined (erotinib and celecoxib). The tumor major axis and short diameter were measured twice a week and after 40 days tissues were collected for immunohistochemical analyses of Bcl-2 and Bax positive cells and Western-blotting analyses for the epidermal growth factor recepto (EGFR), P-EGFR, and cyclooxygenase-2 (COX-2). Tumor size in the combined group was smaller than in the others (p<0.01) and the percentage of Bcl-2 positive cells was fewer in most cases (p<0.01), while that of Bax positive cells was greater than in the erlotinib and celecoxib groups (P>0.05). Western blotting showed decreased expression of P-EGFR and COX-2 with both erlotinib and celecoxib treatments, but most pronouncedly in the combined group (P<0.05). Simultaneous blockage of the EGFR and COX-2 signal pathways exerted stronger growth effects in our human xenotransplanted lung cancer model than inhibition of either pathway alone. The anti-tumor effects were accompanied by synergetic inhibition of tumor cell apoptosis, activation of p-EGFR and expression of COX-2.

• Journal of Society of Preventive Korean Medicine
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• v.8 no.2
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• pp.81-98
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• 2004

1. The cell viability was determined by MTT method. Their cytotoxic activities against three cancer cell lines such as A549, MDA-MB-231 and SNU-C4 cell line were tested. Among them, The methanol extract of Saururus chinensis Baill. showed the strongest cytotoxic effect against SNU-C4 cells. These results suggest that the methanol extract of Saururus Chinensis Baill. possessed a potential antitumorous agent. 2. In vitro the antitoxic activity of ethanol extract of Saururus Chinensis Bail on NIH 3T3 fibroblasts was evaluate by the MTT (3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl-2H-tetra캐lium bromide) and SRB (sulforhodamine B protein) assays. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that Saururus Chinensis Bail extract retains a potential antitoxic activity. 3. Complexation of Cd (II) ion with ligands such as quercetin has been determined by UV-vis spectrophotometric method in tris buffer solution at various pH. It was found that only 1 : 1 Cd-complex is formed by the interaction between catechol moiety in ring B and cadmium [Cd(II)] in aqueous solution. The spectral parameters for Cd-complex were determined by Beer's law at various pH. It has shown that 1 : 1 Cd-complex has a maximum absorbance and red shift by the alkaline pH.