• The Journal of Korean Obstetrics and Gynecology
• /
• v.20 no.3
• /
• pp.1-12
• /
• 2007

Purpose: Uterine leiomyomas (fibroids) are common estrogen-dependent uterine tumors. Houttuynia cordata thunberg has cancer-preventing properties and often used in Chinese medicine. In the present study we used Houttuynia cordata thunberg to determine its effect on cell proliferation and apoptosis in human uterine leiomyoma cells. Methods: Primary cultured human uterine leiomyoma cells were treated with Houttuynia cordata thunberg. Cell viability analysis was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTS) assay and Flow cytometry was performed to ascertain the effects Houttuynia cordata thunberg. Expression of cell cycle related proteins and apoptosis related proteins were evaluated by Western blot analysis. Results: Cell viability was significantly influenced by Houttuynia cordata thunberg treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. Flow cytometric analysis showed that Houttuynia cordata thunberg induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increase in p21 was observed. Houttuynia cordata thunberg treatment of uterine leiomyoma cellsresulted in a concentration-dependent cell death induced via the caspase dependent mechanism. Conclusion: These results suggest that Houttuynia cordata thunberg treatment in uterine leiomyoma cells leads to growth inhibition and induced apoptosis. These results suggest that Houttuynia cordata thunberg will be a promising agent for use in therapeutics agents against human uterine endometrial cancer.

• Molecules and Cells
• /
• v.24 no.2
• /
• pp.276-282
• /
• 2007

Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

• The Journal of the Convergence on Culture Technology
• /
• v.3 no.4
• /
• pp.1-19
• /
• 2017

Death culture continues to lead human history with complementarity in the sense that it is half of life culture. The three sacred gates and the two tombs are connected to the olegil space. In this space, the principle of complementarity in which coexistence exist between life and death is hidden in Jeju culture. It is a question and wait. Contrarily, the opposite is complementary. (Contraria Sunt Complementa Latin) This refers to the relationship of each other in relation to one another and in a mutually dependent relationship. The Jong Nang is used as basic logic in DNA codon of human body as well as communication principle. In addition, there is a pattern of similarity of the linkage between the square oval ear of the grave, the Korean Taegeukguk and 卦, the genome (DNA) 卦. The Jong Nang 'Batdam' and 'Sandam' are conneted like 'black dragon ten thousand', and stone walls are connected to each other as the stone network, which is called as the 'Goendam' (and conventionally is told as the 'Goendang') and is related to the Family networks. The relationship between the Tomb of the ancestors and their offspring is believed to be "Soul Synchronizing the Ancestor to Offspring" and becomes "Change in Future", and Genetic factors in the physical blood are transmitted directly to the children. The DNA RNA was represented by the determinant.

• Environmental Mutagens and Carcinogens
• /
• v.17 no.2
• /
• pp.97-108
• /
• 1997

The drugs or xenobiotics introduced to the body, are detoxified through the process of biotransformation in the body. In this process, most of the insoluble compounds become more polar, soluble and easily excretable. But, parts of introduced materials are metabolized to highly reactive electrophilic carcinogens through activation pathways. These metabolites are toxic and can react with DNA, RNA and proteins which are nucleophilic compounds. The objective of this study is to illustrate the aleactivation pathways of two highly reactive epoxide compounds, vinyl carbamate epoxide (VCO) and 2'-(4-nitrophenoxy)oxirane (NPO). They are the ultimate electrophilic carcinogens of ethyl carbamate(urethane) and 4-nitrophenyl vinyl ether, respectively. In this research, we studied the inhibition of the mutagenic activities of VCO or NPO by nuchieophiles [glutahione(GSH) and N-acetylcysteine(NAC)], detoxifying enzymes[epoxide hydrolase and glutathione-S-transferase(GST)] and intracellular organelles (microsomes and cytosol). In addition we also tested the suppression of DNA adducts formation by GSH and NAC. The results are summerized as follow. 1. The microsomes and cytosol which contain epoxide hydrolase and GST, respectively, decreased the mutagenicity of VCO (74% and 95%, respecfivel), and NPO (35% and 93%, respectively). The nucleophilic GSH and NAC decreased the mutagenicity by 86% (VCO) and 80% (NPO), 76% (VCO) and 40% (NPO), respectively. 2. The purified epoxide hydrolase decreased the mutagenicity of two epoxides in a dose-dependent manner, and GSH also decreased the mutagenicity in the presence of GST. 3. Formation of two DNA adducts, 7-(2'-oxoethyi)guanine (OEG) and N2,3-ethenoguanine(EG), were compared in the presence of calf thymus DNA and epoxide (VCO or NPO) in vitro system. The amounts of DNA adducts were decreased in the presence of GSH (25% and 29% in VCO, 32% and 29% in NPO), and NAC (14% and 16% in VCO, 21% and 11% in NPO), respectively. From these results, it is concluded that the ultimate carcinogenic metabolites, VCO and NPO, can be made in the body, but much of them may be inactivated and detoxified by the nucleophilic GSH, NAC and detoxifying enzymes (epoxide hydrolase and GST). Therefore, by these mechanism, the formation of DNA adducts and mutagenic activities of these two epoxides may be lowered in vivo.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.8 no.1
• /
• pp.37-45
• /
• 1975

The present study was ,conducted to evaluate the effects of condensed phosphates on the refeezing damage of Alaska pollack muscle. The fillets were dipped in such solution as 5 and $10\%$ sodium polyphosphate, 1 and $5\%$ mixture of sodium polyphosphate and sodium pyrophosphate (1:1, w/w) for 1 and 5 minutes, respectively, before refreezing. And fillets were frozen at $27^{\circ}C\~28^{\circ}C$ and stored for 15 days at $-18^{\circ}\~-20^{\circ}$. The degree of denaturation was estimated by determining amounts of drip relased, content of total solids, nitrogen, and DNA in the drip an cooking-weight-loss. Phosphorus absorbed in the muscle was also determined. Phosphorus absorbed in the fillets treated with loft solution of sodium polyphosphate for 5 minutes amounted to 101 mg/100g muscle as $P_2O_5$. The absorption was dependent on tile concentration of treating solution rather than on the dipping time. The increase of phosphorus absorption seemed to affect to reduction of drip. Among the treating conditions, $10\%$, 5 minutes and $10\%$ 1 minute with sodium polyphosphate appeared most effective ones on drip reduction. The effect of $5\%$, minutes with the mixture of sodium polyphosphate and sodium pyrophosphate did not show so benefitable effect in refrozen fillets. As a tendency total solids, nitrogen, and DNA in tile drip varied proportionally to the amount of drip released. And the content of DNA was lower than the amount. Treatment, at higher the concentration and longer the dipping time, resulted in the lower cooking-weight-loss and the better quality on organoleptic test of thawed fillets.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.10
• /
• pp.5941-5948
• /
• 2013

Tumor response to antineoplastic drugs is not always predictable. This is also true for bladder carcinoma, a highly recurrent neoplasia. Currently, the combination of cisplatin and gemcitabine is well accepted as a standard protocol for treating bladder carcinoma. However, in some cases, this treatment protocol causes harmful side effects. Therefore, we investigated the roles of the genes TP53, RASSF1A (a tumor suppressor gene) and hMLH1 (a gene involved in the mismatch repair pathway) in cell susceptibility to cisplatin/gemcitabine treatment. Two bladder transitional carcinoma cell (TCC) lines, RT4 (wild-type TP53) and 5637 (mutated TP53), were used in this study. First, we evaluated whether the genotoxic potential of cisplatin/gemcitabine was dependent on TP53 status. Then, we evaluated whether the two antineoplastic drugs modulated RASSF1A and hMLH1 expression in the two cell lines. Increased DNA damage was observed in both cell lines after treatment with cisplatin or gemcitabine and with the two drugs simultaneously, as depicted by the comet assay. A lack of RASSF1A expression and hypermethylation of its promoter were observed before and after treatment in both cell lines. On the other hand, hMLH1 downregulation, unrelated to methylation status, was observed in RT4 cells after treatment with cisplatin or with cisplatin and gemcitabine simultaneously (wild-type TP53); in 5637 cells, hMLH1 was upregulated only after treatment with gemcitabine. In conclusion, the three treatment protocols were genotoxic, independent of TP53 status. However, cisplatin was the most effective, causing the highest level of DNA damage in both wild-type and mutated TP53 cells. Gemcitabine was the least genotoxic agent in both cell lines. Furthermore, no relationship was observed between the amount of DNA damage and the level of hMLH1 and RASSF1A expression. Therefore, other alternative pathways might be involved in cisplatin and gemcitabine genotoxicity in these two bladder cancer cell lines.

• Journal of Life Science
• /
• v.17 no.4 s.84
• /
• pp.522-528
• /
• 2007

In the present investigation, we studied the modulating effects of caffeic acid, chlorogenic acid, and (-)-epigallocatechin-3-gallate(EGCG) on the methylation status of promoter regions of cell cycle regulator, p16, in human breast cancer T-47D cells. We demonstrated that treatment of T-47D cells with caffeic acid, chlorogenic acid, or EGCG partially inhibited the methylation status of the promoter regions of p16 genes determined by methylation-specific PCR. In contrast, unmethylated p16 genes were increased with the treatment of T-47D cells with $20{\mu}M$ of caffeic acid or chlorogenic acid for 6 days. Treatment of T-47D cells with 5, 20 or $50{\mu}M$ of EGCG increased the unmethylation status of p16 gene up to 100%, and the methylation-specific bands of this gene were decreased up to 50% in a concentration-dependent manner. The finding of present study demonstrated that coffee polyphenols and EGCG have strong inhibitory effects of the cellular DNA methylation process through increased formation of S-adenosyl-homocysteine(SAH) during the catechol-O-methyltransferase (COMT)- mediated O-methylation of these dietary chemicals or an direct inhibition of the DNA methyltransferases. In conclusion, various dietary polyphenols could reverse the methylation status of p16 gene in human breast T-47D cells.

• The Korean Journal of Zoology
• /
• v.34 no.2
• /
• pp.148-158
• /
• 1991

The purpose of this research was to establish the culture condition for dissociated submandibu -lar gland (SG) cells. After trypsin digestion of SG from 3-4 weeks old mice, dissociated cells were cultured in 1OO/o fetal bovine serum-Dulbecco's modified Eagle's medium (FBS-DME) or 0.5-2% low protein serum replacement-DME (LPSR-DME) on plastic surface to form monolayer. The effects of FBS, LPSR and hormones on the growth and function of cultured SG cells were examined. SG cells dissociated by enzyme were successfully cultured and were characterized as epithelial-like cells by light and electron microscope. The maximal DNA synthesis of cultured SG cells was achieved by DME containing 5-10% FBS. The same results were obtained when the effects of LPSR on cell proliferation were examined up to a LPSR concentration of 2%. SG cells cultured in 20/o LPSR-DME expressed a population doubling time of 42.5 hrs and a saturation density of 1.2 $\times$10 5cell/cm$^2$. Dihydrotestosterone (DHT) in medium did not influence on the DNA synthesis of the cultured SG cells, but stimulated protein synthesis of the SG cells. Thyroxine (T4) stimulated protein synthesis of the SCI cells markedly in a dose-dependent fashion. EGF secretion by the cultured SG cells increased significandy by DHT and or T4 trearment. This finding indicated that secretion of EGF by the SG cells was under the control of the hormones such as androgen and thyroid hormones. It seems to be that the culture condition described here can be used as a useful tool for further research on the SCI cells.

• Journal of Marine Bioscience and Biotechnology
• /
• v.2 no.4
• /
• pp.234-237
• /
• 2007

Calmodulin (CaM) is a $Ca^{2+}$-binding protein essential for biological functions mediated through $Ca^{2+}$-dependent mechanism. A cDNA clone for CaM was isolated from a cDNA library of olive flounder Paralichthys olivaceus. The CaM cDNA concists of 782 bp and encodes a polypeptide of 149 amino acids with four $Ca^{2+}$-binding motifs EF-hands (EF-I, EF-II, EF-III, and EF-IV). The deduced amino acid sequence of CaM shows 97-100% amino acid sequence identity to other CaM sequences. Semi-quantitative PCR analysis revealed that the CaM transcription was began during early development and the CaM mRNA is expressed highly in brain and intestine, and moderately in kidney, gill, and eye of healthy olive flounder. Taken together, CaM may be necessary for early olive flounder development and that it may have a part in homeostasis.

• Korean Journal of Plant Resources
• /
• v.37 no.2
• /
• pp.101-109
• /
• 2024

In this study, we investigated the antioxidant effects and active ingredients of Gynura Procumbens extracts obtained through various extraction methods for the development of natural-based cosmetics and pharmaceutical materials. The contents of compounds, total flavonoids, chlorogenic acid, and caffeic acid were compared at different concentrations, revealing the highest content of active ingredients in the 100% ethanol extract. Antioxidant activity assessed by DPPH and ABTS radical scavenging assays showed a concentration-dependent increase in antioxidant activity with the ethanol concentration. Additionally, we validated the DNA damage inhibition and anti-inflammatory effects of Gynura Procumbens extracts through UVB irradiation on Hs68 cell models. The 100% ethanol extract demonstrated significant inhibition of the expression of p-p53, γ-H2AX, iNOS, and COX-2 induced by UVB, indicating its potential in alleviating photodamage effects. Consequently, the efficient extraction of Gynura Procumbens for skin functional material development was confirmed, suggesting the suitability of ethanol or alcohol-based solvents.