• Journal of Oral Medicine and Pain
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• v.34 no.3
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• pp.261-276
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• 2009

Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.57 no.4
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• pp.343-352
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• 2012

This experiment was carried out to report some data such as survival rate, tetraploid production efficiency, and agronomic characteristics of offspring from the induced tetraploid by the colchicine treatment in rye. The colchicine was soaked with 0.05%, 12 hours in dark condition and at two growth stages (green seed and 2nd leaf stage) in diploid ryes. Flow cytometry (FC) was proved to be efficient and rapid tool for screening ploidy levels in rye, showing around 40 to 60 in DNA amount (DAP1) corresponding to diploid and 80 to 110 tetraploid. There were 18.5% of survival rate at green seed treatment and 78% at 2nd leaf stage in average of two rye cultivars, Gogu and Jogreen, but in reverse 50.9% and 1.1% in the ratio of tetraploid to total tillers among the plants survived, respectively, resulting in 9.42% of tetraploid production rate in green seed treatment and 0.86% at 2nd leaf stage, respectively. In green seed treatment, there were 33% of survival rate in Gogu, 4% in Jogreen in 1st year, but 56% in Gogu, 21% in Jogreen and 49% in Charmgreen, respectively. The rate of tetraploid to total spikes among survived was 53.7% in Gogu, 32.4% in Jogreen, and 50.9% in average in 1st year, and 64.1% in Gogu, 51.5% in Jogreen, 60% in Charmgreen, and 60.5% in average in 2nd year. In green seed treatment, tetraploid production rate (survival rate ${\times}$ tetraploid ratio ${\times}$ 100) was 17.7% in Gogu and 1.3% in Jogreen and 9.42% in average in 1st year, and 35.9% in Gogu, 10.8% in Jogreen, 29.4% in Charmgreen, and 25.4% in average of three diploid rye cultivars. By the colchicine treatment with 0.05% for 12 hours in Gogu and Jogreen, 35 tetraploid plants were obtained and they produced 2,673 seeds with 148 spikes. There were 3.3-4.4 in the number of spikes per plant, 15.6-18.3 in grain number per spike, and 37.6 g in Gogu and 46.8 g in Jogreen in the 1,000-grain weight.

• Sim-seong Yeon-gu
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• v.26 no.1
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• pp.67-88
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• 2011

Pungsu is an ancient logic that systemizes geography based on the Yin-Yang and The Five Elements Principle. It is defined as the unique and highly systemized ancient Eastern art of selecting auspicious sites and arranging harmonious structures such as graves, houses, and cities on them by evaluating the surrounding landscape and cosmological directions. Pungsu helps allegedly one improve life by receiving vital energy(Shengqi, 生氣)-energy flow that flows under the ground. It is traditional belief that the living lead their lives on the ground, indirectly receiving the energy coming out of the ground, whereas the dead are buried under the ground, allowing them to directly absorb energy from the ground, which makes Shengqi the dead receive bigger and more obvious than that the living receive. This energy absorbed by the dead from the ground was believed to be passed on to their descendants. This phenomenon is called "Induction of vital energy between Ancestors and Descendants". People searched for the sites which were believed to contain rich and positive vital energy flow. They also tried to bury their ancestors under such sites hoping to receive the Shengqi coming from underground which they believed would help them thrive and prosper. The efforts to locate the sites which have the most vital energy, auspicious sites or "Bright Yard(明堂)", are easily observed in Asia including China and Korea. The ultimate goal of searching for auspicious sites lies in human(whether alive or dead) receiving vital energy from the nature to enjoy happy lives. In choosing a place to live or to bury their dead ancestors, people tried to understand the energy flow of the site considering the factors related with mountain, water, and direction. If we take a closer look into the methods of finding auspicious sites, we can see that people have tried to see the outer conditions of lands, mountains and waters within the perfect harmony if possible. Auspicious site or Bright Yard is the site with those elements in perfect order and harmony, that is, it is the place which derives the most vital energy from the best order and harmony of nature. As this shows, an auspicious site symbolizes totality-the Self, and it seems to be projected to the land. It is believed to be an attempt that the reason why we try to find auspicious sites to internalize the totality that we projected to the outer world. Therefore, this auspicious site is what our foremost values, symbol of the Self, such as harmony, equilibrium, perfection, and uniqueness are reflected to the land. Through the process of finding such a site, we try to gain totality of psyche.

• Journal of Life Science
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• v.21 no.7
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• pp.961-968
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• 2011

The formation of reactive lipid aldehydes, 4-hydroxynonenal (HNE) is shown to be derived from fatty acid hydroperoxides through the oxidative process. Among its known effects in cytotoxicity, HNE has been implicated in apoptotic cell death. To delineate its putative role as a potential mediator, we investigated the mechanism by which HNE induces apoptosis of endothelial cells (ECs). The anti-proliferative effects of HNE were tested through MTT assay after exposure to various concentrations ($5\sim15\;{\mu}M$) of HNE. We observed apoptotic bodies with propidium iodide staining, and measured the HNE induction of endothelial apoptosis by flow cytometry assay. We observed that cells exposed to HNE for 24 hr resulted in increased poly(ADP-ribose) polymerase cleavage and up-regulation of Bax. Data on the HNE action strongly indicated the involvement of reactive species, namely, intracellular ROS, nitrite, and peroxynitrite. To obtain evidence on the implication of ROS and peroxynitrite in HNE-induced apoptosis, a ROS scavenger, N-acetylcysteine (NAC), and a peroxynitrite scavenger, penicillamine, were tested. Results clearly indicate that the induction of apoptosis by HNE was effectively inhibited by NAC and penicillamine. Based on the present data, we conclude that the endothelial apoptosis induced by HNE involves both ROS generation and peroxynitrite activity. Our new data could lead to a redefinition of HNE action on apoptosis in ECs.

• Korean Journal of Plant Resources
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• v.26 no.2
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• pp.284-288
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• 2013

This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.

• Parasites, Hosts and Diseases
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• v.55 no.4
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• pp.439-444
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• 2017

The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of $IFN-{\gamma}$, $TNF-{\alpha}$, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. $Foxp3^+Treg$ were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs ($5.2{\pm}0.4$) was significantly higher than the control ($3.6{\pm}0.5$) (P<0.05). The levels of IL-10 ($55.3{\pm}2.2$ vs $35.2{\pm}3.2$), IL-17 ($52.9{\pm}3.8$ vs $41{\pm}1.8$), $IFN-{\gamma}$ ($44.8{\pm}4.8$ vs $22.3{\pm}2.3$) and $TNF-{\alpha}$ ($71.1{\pm}5.8$ vs $60.1{\pm}3.3$) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.726-733
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• 2005

Histone deacetylase (HDAC) inhibitors target key steps of tumor development. They inhibit proliferation, induce differentiation and/or apoptotic cell death, and exhibit potent antimetastatic and antiangiogenic properties in cancer cells in vitro and in vivo. Although they are emerging as a promising new treatment strategy in malignancy, how they exert their effect on human non-small cell lung cancer cells is as yet unclear. The present study was undertaken to investiate the underlying mechanism of a HDAC inhibitor trichostatin A (TSA)-induced growth arrest and its effect on the cell cycle control gene products in a human lung carcinoma cell line A549. TSA treaoent induced the growth inhibition and morphological changes in a concentration-dependent manner. Treatment of A549 cells with TSA resulted in a concentration-dependent increased G1 (under 100 ng/ml) and/or G2/M (200 ng/ml) cell population of the cell cycle as determined by flow cytometry Moreover, 200 ng/ml TSA treatment significantly induced the population of sub-G1 cells (23.0 fold of control). This anti-proliferative effect of TSA was accompanied by a marked inhibition of cyclins, positive regulators of cell cycle progression, and cyclin-dependent kinases (Cdks) expression and concomitant induction of tumor suppressor p53 and Cdk inhibitors such as p21 and p27 Although further studies are needed, these findings provide important insights into the possible molecular mechanisms of the anti-cancer activity of TSA in human lung carcinoma cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.1
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• pp.41-51
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• 2008

Mucoepidermoid carcinoma (MEC) is the most common malignant salivary gland tumor which compromises about 6$\sim$8% of all tumors followed by the adenoid cystic carcinoma (ACC) and adenocarcinoma. Most deaths from salivary carcinomas are caused by recurrent or metastatic lesions that are resistant to conventional therapy. Therefore, knowledge of cellular properties and tumor-host interactions that influence the vascular metastasis is important for the design of more effective therapy of salivary carcinomas. Neoangiogenesis is essential for tumor growth, which is postulated to be fundamentally dependent on the induction of stromal neovascularization. However, how neovascularization takes place in live tissue has not been fully established, especially in recruitment and differentiation of endothelial cells in the salivary gland tumors. Vascular endothelial growth factor (VEGF) is a heparin-binding, dimeric polypeptide growth factor known to exert its mitogenic activity specifically on endothelial cells. VEGF has been shown th be directly involved in angiogenesis, which in essential for the pathogenesis of many solid tumors. von Willebrand factor (vWF) is a large multimeric protein synthesized by megakaryocytes and endothelial cells that enable platelets to adhere to exposed subendothelium and, as well, to respond to changes in the blood flow. Recent studies suggest that increased levels of vWF correlate with progression of disease, metastasis, or survival time and thus may have a prognostic significance. vWF is explained as an acute phase proteins which is increased in cancer or as a result of increased endothelial cell synthesis associated with tumor-induced angiogenesis. Due to adhesive properties of vWF, its increased concentrations may also contribute metastasis of tumor. In this study, we determined the mRNA expression of VEGF and vWF in salivary ACC, MEC and pleomorphic adenoma by in situ hybridization. As a result, stronger expression of VEGF and vWF was seen in salivary ACC and MEC which has more invasive nature than the salivary benign tumor.

• Imaging Science in Dentistry
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• v.31 no.3
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• pp.135-143
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• 2001

Purpose : To evaluate the effect of all-trans-retinoic acid on radiosensitivity and radiation-induced apoptosis in NHOK, HEp-2 and FaDu cell lines. Material and Methods: We measured the changes in survival fraction at 2 Gy (SF2), α and β after treatment of retinoic acid (1μM) prior to irradiation with doses of 2, 4, 6 and 10 Gy and correlated the radiosensitizing effect of retinoic acid with them. Also, apoptosis induction was assayed with the flow cytometry on days 1,2, 3, 4 and 5 after irradiation (2, 10 and 20 Gy) combined with retinoic acid. Results and Conclusion: SF2 values for NHOK, HEp-2 and FaDu cell lines were 0.54, 0.64 and 0.41, respectively and the cell line of FaDu was the most radiosensitive. For cell lines of NHOK and HEp-2, pretreatment of cells with retinoic acid resulted in a significant decrease of the SF2 values. The α/β ratios of x-ray survival curve were 8.714 (NHOK), 4.098 (HEp-2) and 11.79 (FaDu). The α/β ratio for NHOK decreased on pretreatment with retinoic acid, whereas those for HEp-2 and FaDu increased. Radiation induced apoptosis in all cell lines but, retinoic acid did not affect the apoptosis.

• Imaging Science in Dentistry
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• v.33 no.3
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• pp.143-149
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• 2003

Purpose: To investigate the radiosensitivity of the normal human oral keratinocytes (NHOK), and the effect of irradiation on cell cycle and protein expression. Materials and Methods: To evaluate the radiosensitivity of NHOK, the number of colonies and cells were counted after irradiation and the SF2 (survival fraction at 2Gy) value, and the cell survival curve fitted on a linear-quadratic model were obtained. LDH analysis was carried out to evaluate the necrosis of NHOK at 1, 2, 3, and 4 days after 2, 10, and 20 Gy irradiation. Cell cycle arrest and the induction of apoptosis were analyzed using flow cytometry at 1, 2, 3, and 4 days after 2, 10, and 200y irradiation. Finally, proteins related cell cycle arrest and apoptosis were analysed by Western blot. Results: The number of survived cell was significantly decreased in a dose-dependent manner. The cell survival curve showed SF2, α, and β values to be 0.568, 0.209, and 0.020 respectively. At 200y irradiated cells showed higher optical density than the control group. After irradiation, apoptosis was not observed but G2 arrest was observed in the NHOK cells. 1 day after 10 Gy irradiation, the expression of p53 remained unchanged, the p2l/sup WAF1/Cipl/ increased and the mdm2 decreased. The expression of bax, bcl-2, cyclin B1, and cyclin D remained unchanged. Conclusion: These results indicate that NHOK responds to irradiation by G2 arrest, which is possibly mediated by the expression of p21/sup WAFl/Cipl/, and that cell necrosis occurs by high dose irradiation.