• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.339-353
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• 1994

Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.2
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• pp.200-207
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• 2015

Various natural products or their derivatives, mostly originating from plants, fungi, and bacteria, have been exploited as therapeutic drugs to treat various human diseases. In addition to previously explored organisms, research on natural compounds has now expanded into unexamined living organisms in order to identify novel bioactive substances. Here, we determined whether or not the larval form of the mealworm beetle Tenebrio molitor, a species of darkling beetle, contains cytotoxic substances that exclusively affect cancer cell viability. Ethanol extract and its solvent partitioned fractions, hexane and ethyl acetate fractions, showed anticancer effects against various human cancer cells derived from the prostate (PC3 and 22Rv1), cervix (HeLa), liver (PLC/PRF5, HepG2, Hep3B, and SK-HEP-1), colon (HCT116), lung (NCI-H460), breast (MDA-MB231), and ovary (SKOV3). Cell death induced by the fractions was a mix of apoptosis, necrosis, and autophagy. The hexane fraction was administered intraperitoneally to nude mice bearing a hepatocellular carcinoma SK-HEP-1 and showed inhibition of tumor growth in vivo. Therefore, we concluded that worm extracts contain cytotoxic substances, which can be enriched by proper fractionation protocols, and further separation and purification could lead to the identification of novel molecules to treat human cancers.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.921-927
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• 2001

This study was performed to investigated the effects on the cytotoxicity and antigenotoxicity of Cordyceps militaris extracts on the human cancer cell lines. The ethanol extract and five fractions which were hexane, chloroform, ethylacetate, butanol and aqueous were screened for crytotoxicity on human lung carcinoma(A549). human breast adenocarcinoma (MCF-7) human epitheloid carcinoma(HeLa), human fibrosarcoma(HT1080) human hepatocellular carcinoma(Hep3B), human gastric carcinoma(KATOIII) and chronic myelogenous leukemia(K562) cell by SRB and MTT assays. The results showed that growth inhibition rates of the human cancer cell in the presence of Cordyceps militaris were inhibited with increasing concentration of the extract. The ethanol extract from Cordyceps militaris had strong inhibitory effects in1 mg/mL treatment by SRB assay , showing 89.4%, 85.7%, 72.9% and 65.5% inhibition in HT1080, HeLa, Hep3B and A549, respectively. The treatment of 1 mg/mL hexane fraction by SRB assay had the strongest cytotoxicity with 97.0% on HT1080 followed by MCF-7(92.9%) and HeLA(90.3%). The inhibition ration on KATOIII by MTT assay was much higher in the butanol (83.7%) and aqueous (80.4%) than in the ethanol extract (61.5%) And also, K562 showed similar tendency with KATOIII. The effects of Cordyceps militaris extracts on the frequencies of micronucleated polychromatic erythrocytes (MNPCEs) induced by N-methyl-N-nitro-N-nitrosoguanidime(MNNG) were investigated in the bone-marrow cells of ICR male mice. The amount of 10, 20, 40 and 80 mg/kg of each extract were administered to animals immediately after injection of MNNG, and the exposure time was 36 hours. Significant reductions(p<0.05) with 39.7%, 52.7%, 71.4% and 83.9% were observed in the frequencies of MNPCE when 10, 20, 40 and 80 mg/kg of the hexane fraction of Coryceps militarus extracts were given to the mice.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.10
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• pp.1295-1301
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• 2009

This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effect of Codonopsis lanceolata (CL). CL was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of CL extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. CL extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of CL (200 ${\mu}g$/plate) showed approximately 72.1% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 69.6% and 67.0% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of CL extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (HepG2), human breast adenocarcinoma (MCF-7), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL CL ethyl acetate fraction had the highest cytotoxicity of 74.5%, 70.7% and 80.3% against HeLa, MCF-7 and A549 cells, respectively. In contrast, the extract and its fractions showed only 2$\sim$31% cytotoxicity for a normal human kidney cell line (293). In vivo anticancer effect of CL extract was tested using Balb/c mice transplanted sarcoma-180 cells. CL ethyl acetate fraction showed the highest inhibition rate of 56.4% at the 50 mg/kg concentration.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.6
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• pp.918-922
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• 2016

The objective of this study was to determine the antioxidant and anti-proliferative activities of methanol, ethanol, acetone, and ethyl acetate extracts from oats (Avena sativa L.). Total polyphenol contents of extracts were analyzed by Folin-Ciocalteu assay. The antioxidant activities of extracts were determined by 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities and reducing power. The anti-proliferative activities of colon (HCT116), lung (NCI-H460), and breast (MCF7) cancer cells were investigated. Among solvents, methanol extract showed the highest amount of total polyphenols, which was 8.2 mg gallic acid equivalents/g residue. High levels of ABTS radical [12.1 mg Trolox equivalent antioxidant capacity (TEAC)/g residue] and DPPH radical (4.4 mg TEAC/g residue) scavenging activity and reducing power ($A_{700}=0.39$) were found in methanol extracts. Moreover, methanol extracts indicated higher anti-proliferative activities against HCT116 (69.5%), NCI-H460 (75.2%), and MCF7 (84.8%) cells compared with other extracts. The results show that methanol was the best solvent for extraction of antioxidant and anti-proliferative compounds from oats. Moreover, notable antioxidant and anti-proliferative activities of oats could have significant health benefits.

• Journal of Life Science
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• v.27 no.4
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• pp.456-463
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• 2017

The purpose of this study was to investigate the effect of 3 types of medicinal herbs (Glycyrrhizae radix, Astragali radix and Dioscorea rhizoma) extracts on estrogen-like activities, proliferation and differentiation in osteoblast. Human breast cancer cell line MCF7 was transfected using an estrogen responsive luciferase reporter plasmid for measure the estrogen-like activity. Estrogen-like activities of extracts were in the range of 1.11~5.73 fold to that of negative control. The extract of G. radix showed the strongest estrogen-like activities. The estrogen-like activities of 50 and $500{\mu}g/ml$ extracts of G. radix were similar to that of $10^{-8}$ and $10^{-7}$ M standard solution ($17{\beta}-estradiol$), respectively. G. radix extract showed no cytotoxicity against osteoblast MC3T3-E1 cells at $1{\sim}1,000{\mu}g/ml$. The extract of A. radix showed no significant proliferation of osteoblast. However, the extract of G. radix and D. rhizome showed maximum 148% and 133% proliferation effects. The extract of G. radix also increased alkaline phosphatase activity and the maximum was 122% at $100{\mu}g/ml$ compared to that of control. The nodule formation by the method of the Alizarin red S staining increased compared to control. These results suggest that G. radix is able to perform the bone formation and prevent osteoporosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.3
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• pp.269-275
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• 2008

The avocado is a widely grown and consumed fruit that is high in nutrients and low in calories, sodium, and fats. In this study, antioxidant activities and induction of apoptosis by methanol extracts from sarcocarp, seed and peel of avocado were investigated in vitro. Contents of total polyphenols in methanol extracts from sarcocarp, seed and peel were 13.89, 137.12 and $223.45{\mu}g/mg$ respectively. Radical-scavenging activities of the methanol extracts were examined by using ${\alpha},{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radicals and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assay. The methanol extracts from the peel of avocado showed higher scavenging activities against DPPH, ABTS than those from sarcocarp and seed. Apoptosis in MDA-MB-231 cells mediated by the methanol extracts of avocado was associated with the increase of activation of caspase-3 and caspase-3 target protein, PARP. Therefore, with more researches on identification and action mechanism of active compounds, the methanol extracts from peel and seed of avocado is expected to be a natural source for the developments of functional food and medical agents to prevent human breast cancer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.4
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• pp.416-421
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• 2008

This study was performed to determine the antimutagenic and anticytotoxic effects of soybean paste (doenjang) added deep sea water salt and see tangle in Salmonella Typhimurium TA98, TA100 and human cancer cell lines. In the Ames test, methanol extract of doenjang did not exhibit any mutagenicity but showed substantial inhibitory effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The methanol extracts of doenjang ($200{\mu}g$/plate) added deep sea salt and see tangle (doenjang C) showed approximately 89.1% and 70% inhibitory effect on the mutagenesis induced by MNNG and 4NQO against TA100 strain, whereas 84.4% inhibitions were observed on the mutagenesis induced by 4NQO against TA98 strain. The cytotoxic effects of doenjang methanol extracts against the cell lines with human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human gastric carcinoma (AGS), human lung carcinoma (A549) and human breast adenocarcinoma (MCF-7) were inhibited with the increase of the extract concentration. The treatment of 1.0 mg/mL doenjang C of methanol extracts showed strong cytotoxicities of 71%, 74.4%, 66.2%, 77.3%, and 71.2% against HeLa, Hep3B, AGS, A549, and MCF-7, respectively. In contrast 1 mg/mL treatment of doenjang C methanol extracts had only $10{\sim}40%$ cytotoxicity on normal human embryonal kidney cell (293). Doenjang methanol extract inhibited significantly the tumor growth in mice injected sarcoma-180 cells. Especially, doenjang C methanol extract showed an inhibition of tumor cell activity of 33% by the administration of 25 mg/kg methanol extracts.