• Microbiology and Biotechnology Letters
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• v.50 no.3
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• pp.387-394
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• 2022

Verotoxin-1 (VT-1) or Shiga-like toxin 1 (Stx-1) is produced by enterohemorrhagic Escherichia coli (EHEC) and is an AB5 holotoxin with a strong inhibitor of protein synthesis. VT-1 is a type 2 ribosome-inactivating protein (RIP) that has been shown to have cytotoxic and anticancer potential by inducing necrosis, apoptosis, and cell cycle arrest, making it a promising antitumor candidate. Here, we tested the cytotoxicity of VT-1 on CaCo2 and NCM425 cell lines and the results showed that VT-1 was more potent on CaCo2. Morphological changes were also evaluated on the cellular level and the results showed that VT-1 caused a decrease in viable cell count, altered cell membrane permeability, and an increase in total nuclear intensity. On the other hand, VT-1 displayed a lesser impact on mitochondrial membrane potential (MMP) and cytochrome c release. On the expression of caspases 3 and 9, VT-1 exhibited an insignificant effect on both which alongside the mitochondrial membrane potential (MMP) and cytochrome c results, might indicate that CaCo2 suffered from the necrosis process as a mechanism of cell death after exposure to VT-1.

• 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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1089-1097
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• 2004

This study was designed to elucidate the synergistic cytotoxic mechanisms of the cotreatment of Gagamsibjeondaebo-tang (GSD) and AS₂O₃ in human lung cancer cell line, H-460. The combination of GSD and AS₂O₃ synergistically augmented the cytotoxicity of GSD and AS₂O₃ in H-460 cells. The nature of cytotoxicity was revealed as apoptosis which was characterized by chromatin condensation and fragmentaton in DAPI staining. The apoptotic cytotoxicity of GSD and AS₂O₃ was accompanied by the cleavage of PARP. Of note, the expression of pro-apoptotic BclXS protein was increased, but the expressions of Sax and BclXL was not affected in H-460 cells treated with GSD and AS₂O₃. In addition, antioxidant NAC completely blocked the apoptotic death of H-460 cells by GSD and AS₂O₃. In conclusion, this results suggest that the cotreatment of GSD and AS203 induces the synergistic apoptotis of human large cell lung cancer cell line, H-460 via the induction of BclXS and reactive oxygen species (ROS).

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7335-7338
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• 2013

There is increasing evidence that natural killer (NK) cells play an important role in antitumor immunity following dendritic cell (DC) vaccination. Little is known, however, about the optimal stimulation of DCs by epitopes and NK interactions for cytotoxicity in tumors. In this study, DC cells activated by the HPV16E7.49-57 epitope and LPS were co-cultured with NK cells in vitro, and then used ot immunize mice to study CTL activity of TC-1, which constitutively expresses HPV16E6E7, with an LDH release assay. Cytotoxicity in mice immunized with DC loaded with epitope HPVE7.49-57 vaccine co-cultured with NK was enhanced significantly (p<0.01). In conclusion, talk-across between DC and NK cells enhances their functions, also improving cytotoxicity againsttumor cells, suggesting that activated DC-NK by epitopes has potential application for cancer-specific immuno-cellular therapy.

• Journal of Korea Foresty Energy
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• v.20 no.2
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• pp.1-8
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• 2001

The 90% methanol extracts of eight magnoliaceae plants were collected and tested the cytotoxicity against SK-OV-3 and SiHa cells. Also six pure compounds such as magnonol, honokiol, dihydroxybiphenyl ether, linodenine, anonaine, asimilobine which were previously isolated from Magnolia obovata Thunb. were evaluated the cytotoxicities and their mechanism study using the Lactate dehydrogenase assay(LDH) and FACScan analysis system. Of the tested six compounds, magnonol, honokiol, dihydroxybiphenyl ether showed high cytotoxicities against human cancer cell lines, SK-OV-3 and SiHa cells. In addition, one of the plausible mechanisms of their antitumor activities suggested that they could induce the early stage of apoptosis. For the quantitative analysis, the methanol extractives were fractionated with chloroform, ethylacetate, $H_2O$ and then the ethylacetate fraction was chromatographed on silica gel using n-Hexane ; Acetone(4:1, v/v) as eluent. This fraction was subjected for the quantitative analysis in the HPLC system. The result suggested that the methanol extractives of Magnolia obovata Thunb. contained with magnonol, honokiol, dihydroxybiphenyl ether, 0.9%, 0.3% and 0.24%, respectively.

• Herbal Formula Science
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• v.17 no.2
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• pp.73-83
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• 2009

Objective : In vitro proapoptotic effect of Hangbaek-Tang (HBT) has been documented by one of us. In the present study, we aimed to demonstrate in vivo effect of HBT on tumor growth. Methods : In vitro selective cytotoxicity of HBT was examined by enumeration of viable cell numbers using BC3A mouse leukemic cells and normal spleen cells. In vivo effect of HBT (25 and 50 mg/mouse) on tumor growth was assayed using BC3A cells innoculated subcutaneously in the flank. Annexin-V apoptosis assay and PI staining was performed to determine the effective serum factor in HBT-treated mice. Leukocyte recruitment into peritoneum were analyzed by microscopy with a stained cytosmear of peritoneal lavage fluid. Results : HBT exhibited in vitro selective cytotoxicity to leukemic cells and did not show any toxicity on immune organs. In vivo i.p. administration of HBT induced significant reduction in tumor growth but not complete regression. Sera obtained from HBT-treated mice strongly inhibited BC3A cell growth in vitro and were revealed to markedly enhance apoptosis and accompanying cell death, when compared to those from PBS-treated mice. Abundant extravasation of leukocytes, especially neutrophils, into peritoneum was observed in HBT-treated mice. Conclusions : HBT causes leukemic, BC3A cell death in vivo via apoptosis as well as in vitro, for which functional involvement of leukocytes is suggested.

• Journal of Ginseng Research
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• v.44 no.5
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• pp.673-679
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• 2020

Background: Panax notoginseng saponin (PNS) is the extraction from the roots and rhizomes of Panax notoginseng (Burk.) F. H. Chen. PNS is the main bioactive component of Xuesaitong, Xueshuantong, and other Chinese patent medicines, which are all bestselling prescriptions in China to treat cardiocerebrovascular diseases. Notoginsenoside R₁ and ginsenoside Rg₁, Rd, Re, and Rb₁ are the principal effective constituents of PNS, but a systematic research on the rare saponin compositions has not been conducted. Objective: The objective of this study was to conduct a systematic chemical study on PNS and establish the HPLC fingerprint of PNS to provide scientific evidence in quality control. In addition, the cytotoxicity of the new compounds was tested. Methods: Pure saponins from PNS were isolated by means of many chromatographic methods, and their structures were determined by extensive analyses of NMR and HR-ESI-MS studies. The fingerprint was established by HPLC-UV method. The cytotoxicity of the compounds was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5 -diphenyltetrazolium bromide assay. Results and Conclusion: Three new triterpenoid saponins (1-3) together with 25 known rare saponins (4-28) were isolated from PNS, except for the five main compounds (notoginsenoside R1 and ginsenoside Rg1, Rd, Re, and Rb1). In addition, the HPLC fingerprint of PNS was established, and the peaks of the isolated compounds were marked. The study of chemical constituents and fingerprint was useful for the quality control of PNS. The study on antitumor activities showed that new Compound 2 exhibited significant inhibitory activity against the tested cell lines.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.28 no.1
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• pp.271-283
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• 1998

The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for human squamous cell carcinoma KB cell line after radiation exposure and/or administration of antitumor drugs. 2, 4, 6, 8, 10Gy were irradiated at a dose rate of 210cGy/min using /sup 60/Co Irradiator ALDORADO 8. After irradiation, KB cell lines (3×104cells/ml) were exposed to 2㎍/ml of bleomycin or cisplatin for 1 hour. The viable cells were determined by MTT assay for each radiation dose and/or each drug at the 4th day. And they were compared to control values. The obtained results were as follows : 1. The slope of the surviving curve after irradiation of 2, 4, 6, 8, 10Gy on KB cell line was relatively steep. 2. There was no significant difference between the cytotoxicity of bleomycin compared to control group. But, there was significant difference between the cytotoxicity of cisplatin compared to control group. And the cytotoxicity of cisplatin was greater than that of bleomycin on KB cell line. 3. There were significant differences of surviving fractions after irradiation of 2Gy and 10Gy with 2㎍/ml of bleomycin compared with the groups of irradiation only on KB cell line. 4. There were significant differences of surviving fractions after irradiation of 2, 4, 6, 8, 10Gy with 2㎍/ml of cisplatin compared with the groups of irradiation only on KB cell line. 5. There was significant difference of surviving fraction between groups after irradiation of 10Gy with 2㎍/ml of bleomycin and cisplatin.

• The Korean Journal of Food And Nutrition
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• v.16 no.1
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• pp.15-21
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• 2003

The purpose of this study is to observe the effect of protein-bound polysaccharide (PBP) on proliferation of Th1 cells and cytotoxicity of cancer cell. Mushrooms (Ganoderma lucidum, Agaricus blazei, Lentinus edodes, Coriolus versicolor and Phellinus linteus) were fractionated by 100$^{\circ}C$ hot water for 3hr. PBP was stimulated and proliferated Th1 cells most at 10 mg/$m\ell$ concentration and the percentage of cell proliferation was 40%. It was estimated cytotoxicity of PBP against 7 kinds of cancer cell lines. Antitumor activities of Agaricus blazei against P388D1 and L1210 (tumor cell lines) were 2.4% and 39.7% survival rate, and Lentinus edodes was 48.4% and 52.5% survival rate, respectively. PBP mixtures of Agaricus and Lentinus edodes prolonged (27∼40%) significantly the survival rate of mice intraperitoneally implanted with sarcoma 180.

• The Korean Journal of Mycology
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• v.18 no.2
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• pp.58-69
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• 1990

ABSTRACT: In order to find physiologically active components from higher fungi, hot-water soluble components were extracted from the basidiocarps of Ganoderma lucidum. The extract was purified and separated by DEAE cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions were designated CR, IN, IA, GL and GH. Fraction GL showed the highest antitumor activity among the fractions and its molecular weight was found to be 47 KD. The tumor inhibition ratio of Fr. GL was 81 % at the dose of peritoneal administration of 20 mg/kg/day for 10 days in mice. Chemical analysis of this fraction showed 82% polysaccharide, 8% protein and 0.9% hexosamine. The polysaccharide moiety consisted of 63% glucose, 27% galactose, 7% mannose and 3% fucose. Fraction IN was found to increase the amount of superoxide anion in activated macrophages to 1.6-fold and the number of plaques in hemolytic plaque assay to 6-fold, respectively. These results indicate that the antitumor activity was exerted through immunopotentiation, but not through direct cytotoxicity against the tumor.