• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.432-441
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• 2016

Machaerium cuspidatum, a canopy liana, is a species of genus legume in the Fabaceae family and contributes to the total species richness in the tropical rain forests. In the present study, we investigated the antioxidative and anti-cancer effects of M. cuspidatum and its mode of action. The methanol extract of M. cuspidatum (MEMC) exhibited anti-oxidative activity with an $IC_{50}$ value of $1.66{\mu}g/ml$, and this was attributable to its 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity. MEMC also exhibited a cytotoxic effect and induced morphological changes in a dose-dependent manner in several cancer cell lines including human lung adenocarcinoma A549 cells, human hepatocellular carcinoma HepG2 cells, and human colon carcinoma HT29 cells. Moreover, MEMC treatment induced the accumulation of subG1 population, which is indicative of apoptosis in A549 and HepG2 cells. MEMC-induced apoptosis was confirmed by the increase in Annexin V-positive apoptotic cells and apoptotic bodies using Annexin-V staining and DAPI staining, respectively. Further investigation showed that MEMC-induced apoptosis was associated with the increase in p53 and Bax expression, and the decrease in Bcl-2 expression. In addition, MEMC treatment led to proteolytic activation of caspase-3, 8, and 9 and degradation of poly-ADP ribose polymerase (PARP). Taken together, these results suggest that MEMC may exert a beneficial anti-cancer effect by inducing apoptosis via both the extrinsic and intrinsic pathways in A549 and HepG2 cells.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10407-10412
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• 2015

Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.7
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• pp.949-955
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• 2011

[ ${\beta}$ ]Glucan is a polysaccharide expressed on the cell walls of fungi. It is known that ${\beta}$-glucan is recognized by a family of C-type lectin receptors, dectin-1, which is expressed mainly on myeloid immune cells, including macrophages, neutrophils and dendritic cells. Raw 264.7 cells were treated with ${\beta}$-glucan from Schizophyllum commune. ${\beta}$-Glucan was not cytotoxic up to 400 ${\mu}g$/mL as measured by MTT assay. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cells. Treatment with ${\beta}$-glucan for 24 hr significantly increased production of NO and TNF-${\alpha}$ compared with control groups (p<0.05), indicating activation of macrophages. To measure inhibition of breast cancer cell proliferation, MTT assay was performed in MDA-MB-231 cells. Cell viability was significantly decreased in the group treated with 400 ${\mu}g$/mL of ${\beta}$-glucan for 48 hr (p<0.05) compared to the control group. However, tumor volume was decreased in the groups administered 200 ${\mu}g$ of ${\beta}$-glucan/mouse compared to the control group. These results indicate that ${\beta}$-glucan inhibits breast cancer cell growth through the induction of apoptosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1015-1021
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• 2013

Glycyrrhiza inflata Batal, an important species of licorice, is one of the most widely used medicinal plants for over 4000 years. Glycyrrhiza plant species has been well known for its various therapeutic activities such as anti-inflammatory, anti-allergic, and anti-ulcer. The purpose of this study was to determine the effects of Glycyrrhiza inflata Batal ethanol extracts (GBE) on adipocyte and osteoblast differentiation. Mesenchymal C3H10T1/2 cells were treated with sub-cytotoxic doses of GBE, and its effects on adipocyte differentiation were assessed. We found that GBE dose-dependently increased lipid accumulation and also induced the expression of adipocyte markers, such as $PPAR{\gamma}$ and its target genes, aP2, and adiponectin, in C3H10T1/2 cells. Consistently, similar effects of GBE on lipid accumulation were also observed in preadipocyte 3T3-L1 cells that further supports the pro-adipogenic activities of GBE. We also investigated the effects of GBE on osteoblast differentiation of mesenchymal C3H10T1/2 cells. As a results, we found that GBE increased the activity of alkaline phosphatase in a dose-dependent manner and also promoted the expression of osteoblast markers, such as ALP and RUNX2, during osteoblast differentiation of C3H10T1/2 cells. Similar pro-osteogenic effects of GBE were also observed in preosteoblast MC3T3-E1 cells. Finally, our data show that a major bioactive compound found in Glycyrrhiza inflata Batal, licochalcone A (LA) but not glycyrrhizic acid (GA), can mediate the pro-adipogenic and pro-osteogenic effects of GBE. Taken together, this study provides data to show the possibility of GBE and its bioactive component LA as putative strategies for type 2 diabetes and bone diseases.

• Parasites, Hosts and Diseases
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• v.28 no.3
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• pp.143-154
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• 1990

Observations were made on the differences of cell-mediated responses in mice of three infectiorl groups di여erently scheduled in their severity with pathogenic Acanthamoeba culbertseni. Infections were done by dropping $5{\;}{\mu}l$ saline suspension containing $3{\times}10^3,{\;}1{\times}10^4,{\;}or{\;}1{\times}10^5$ trophosoites, respectively. Amoebae were cultured anenically in CGV medium and inoculated into the right nasal cavity of CSH/HeJ mice aging around 6∼8 weeks, under the anesthesia by intraperitoneal injection of secobarbital. Delayed type hypersensitivity (DTH) responses in footpad and blastogenlc responses of mouse spleen cells using ($^3H$)-thymidine and the serum antibody titer were measured up to day 14 after infection, and natural killer cell activities were measured up to day, i after infection. The results obtained in this study were as follows: 1. The mice infected with $3{\times}10^3$ trophosoites showed mortality rate of 17%, and 345 in the mice infected with $1{\times}10^4$ trophozoites and 65% with $1{\times}10^5$ trophozoites. 2. In regard to DTH responses in all experimental groups, the level increased on day 7 and declined on day 14 after infection, but their differences could not be noted between infected and control groups. 3. The blastogenic responses of splenocytes treated with amoeba Iysates and lipopolysaccharides (LPS) showed no difference from the control group. The blastogenic responses of splenocytes treated with concanavalin A were declined significantly in the experimental group as compared with the control group, but the blastogenic responses of splenocytes treated with polyinosinic acid were not different from the control group. There was also no difference among three infected groups. 4. The cytotoxic activity of the natural killer cells was activated on day 1 after infection and declined to the level of control group on day 2 in all experimental groups. On day 5 after infection, the natural killer cell cytotoxicity was significantly suppressed as compared with the control groups. 5. The serum antibody titers of the infected mice increased after day 7, but there was no statistical difference between the three infected groups. In summary of the results, there was no difference in cell-mediated immune responses of three experimental groups scheduled with different infection intensities. But there was a significant difference in cell$.$mediated immune responses between infected and control mice. It is considered that cell-mediated immune responses should be involved in murine model infected with A. culbertsoni.

• Journal of Life Science
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• v.20 no.1
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• pp.103-112
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• 2010

Pemetrexed has demonstrated clinical activity in non-small cell lung cancer (NSCLC) as well as other solid tumors. It transports into the cells via reduced folate carrier (RFC) and is polyglutamated by folypolyglutamate synthetase (FPGS). Pemetrexed directly inhibits several folate-dependent enzymes such as thymidylate synthase (TS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). We investigated the effects of genetic variations and the expression of RFC, FPGS, TS and DHFR enzymes on drug sensitivity to pemetrexed in NSCLC cells. Polymorphisms in RFC, FPGS, and DHFR were genotyped in four NSCLC cells - A549, PC14, HCC-1588, and H226. Real-time RT-PCR and Western blot was performed to evaluate mRNA transcripts and protein of these genes. The cytotoxicity of pemetrexed was measured by SRB assay. In PC14 and H226 cells, increased mRNA expressions of RFC and FPGS were associated with higher cytotoxicity to pemetrexed. 2R/2R genotype of TS and its increased mRNA expression were associated with drug resistance to pemetrexed in A549 cells, whereas 3R/3R genotype in TS with decreased mRNA expression was associated with higher sensitivity in H226 cells. After pemetrexed treatment, an inverse change of DHFR mRNA and protein expression was found. The strongest linkage disequilibrium (LD) was discovered between-1726C>T and -1188A>C SNP of DHFR gene. Our findings suggest the cytotoxic effect of pemetrexed may be associated with genetic polymorphisms and the expression level of genes involved in pemetrexed metabolisms in NSCLC cells.

• Radiation Oncology Journal
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• v.24 no.1
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• pp.51-57
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• 2006

Puroose: We examined whether intratumoral (i.t.) administration of dendritic cells (DCs) into a treated tumor could induce local and systemic antitumor effects in a mouse tumor model. Methods and Materials: C57BL/6 mice were inoculated s.c. in the right and left thighs with MCA-102 fibrosarcoma cells on day 0 and on day 7, respectively. On day 7, the tumors (usually 6 mm in diameter) on the right thigh were heated by immersing the tumor-bearing leg in a circulating water bath at $43^{\circ}C$ for 30 min; thereafter, the immature DCs were i.t administered to the right thigh tumors. This immunization procedure was repeated on days 7, 14 and 21. The tumors in both the right and left thighs were measured every 7 days and the average sizes were determined by applying the following formula, tumor $size=0.5{\times}(length+width)$. Cytotoxicity assay was done to determine tumor-specific cytotoxic T-lymphocyte activity. Results: Hyperthermia induced apoptosis and heat shock proteins (HSPs) in tumor occurred maximally after 6 hr. For the local treated tumor, hyperthermia (HT) alone inhibited tumor growth compared with the untreated tumors (p<0.05), and furthermore, the i.t. administered DCs combined with hyperthermia (HT + DCs) additively inhibited tumor growth compared with HT alone (p<0.05). On the distant untreated tumor, HT alone significantly inhibited tumor growth (p<0.05), and also HT + DCs potently inhibited tumor growth (p<0.001); however, compared with HT alone, the difference was not statistically significant. In addition, HT + DCs induced strong cytotoxicity of the splenocytes against tumor cells compared to DCs or HT alone. Conclusion: HT + DCs induced apoptosis and increased the expression of HSPs, and so this induced a potent local and systemic antitumor response in tumor-bearing mice. This regimen may be beneficial for the treatment of human cancers.

• The Journal of Korean Obstetrics and Gynecology
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• v.29 no.1
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• pp.14-34
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• 2016

Objectives: The objective of this study was to evaluate the effects of cytotoxicity, skin regeneration, anti-wrinkle, whitening and skin moisturizing of Oncheongeum (OCE).Methods: The cytotoxicity of OCE lyophilized aqueous extracts (yield=13.82%) was observed against human normal fibroblast cells and B16/F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also evaluated through the assay of collagen type I synthesis compared to the transformation of the growth factor (TGF)-β1, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays compared to oleanolic acid (OA), and elastase inhibitory effects compared to phosphoramidon disodium salt (PP). In addition, OCE’s whitening effects were measured by a tyrosinase inhibitory assay and melanin formation test in B16/F10 murine melanoma cells compared to arbutin, and skin moisturizing effects were observed through a mouse skin water content test, respectively. Results: No OCE treatment-related cytotoxic effects appeared on human normal fibroblasts and B16/F10 murine melanoma cells. OCE concentration-dependently increased the collagen Type I synthesis on human normal fibroblast cells, and also effectively inhibited hyaluronidase, elastase, collagenase and MMP-1 activities. In addition, OCE inhibited melanin production of B16/F10 murine melanoma cells and activity of tyrosinase. And significant and dose-dependent increases of skin water content were detected in OCE-treated mouse skin compared to vehicle control skins. Conclusions: OCE showed favorable and sufficient effects in skin regeneration, anti-wrinkle, whitening and skin moisturizing in this experiment. But more detail mechanisms and studies on the skin protective efficiency of in vivo are needed with the screening of active biological compounds in individual OCE herbs.

• Journal of Life Science
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• v.31 no.3
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• pp.287-297
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• 2021

Cornus officinalis Siebold & Zucc. is traditionally used as an edible and medicinal plant in many countries in East Asia. Previous studies have shown the pharmacological potential of extracts and components of C. officinalis, but comparative analysis of the composition of the leaf, stem, and fruit extracts has been insufficient to date. In the present study, the content of active antioxidant and anti-inflammatory ingredients was verified in different C. officinalis parts (under-ripe sansuyu, ripe sansuyu, seed, leaf, stem, and dried sansuyu). One active component, morroniside, was high in fruit (under-ripe and ripe sansuyu), while loganin was high in fruit (under-ripe sansuyu) and cornin was high in seeds. Total polyphenol contents were highest in fruit (ripe sansuyu) and flavonoids were highest in leaves. DPPH radical scavenging activity was highest in leaves, followed by seeds and then ripe sansuyu extract. The anti-inflammatory efficacy of leaf extracts of C. officinalis (LCO) was further investigated by measuring their effects on levels of nitric oxide (NO) and the pro-inflammatory cytokines interleukin (IL)-1β and IL-6 in RAW 264.7 macrophages. Non-cytotoxic concentrations of LCO effectively decreased the lipopolysaccharide (LPS)-induced expression of inducible NO synthase, resulting in decreased NO production. LCO also significantly suppressed LPS-induced production and expression of IL-1β and IL-6. Taken together, the present findings suggest that C. officinalis leaves have potential as natural materials for the development of antioxidant and anti-inflammatory agents.

• Korean Journal of Food Science and Technology
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• v.53 no.1
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• pp.46-54
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• 2021

The biological activities of non-edible extracts of asparagus stems and roots were investigated using hot water and ethanol. The highest contents of rutin and total polyphenol were 31.74 mg/g and 20.14 mg GAE/g, respectively, in the stem hot water extract. ABTS and DPPH radical scavenging activities were 541.1±21.0 and 649.5±6.6 ㎍/mL, respectively, in stem hot water extract. All extracts were non-cytotoxic in HepG2 cells, but 200 ㎍/mL stem extracts tended to decrease the viability of RAW 264.7 cells. The highest xanthine oxidase inhibitory activity was 43.68% in the root hot water extract at 200 ㎍/mL. The expression level of MMP-9 was significantly decreased in the asparagus extracts. The highest GGT, AST, and LDH activities showed a concentration-dependent decrease in the stem ethanol extract. In conclusion, the presence of bioactive substances in the non-edible extracts of asparagus was confirmed for the development of extracts with antioxidant, hepatoprotective and anti-gout activities.