• Journal of Convergence for Information Technology
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• v.11 no.8
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• pp.201-206
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• 2021

The aim of this study was to examined the dermatoxicity of ferrous chloride (FeCl2) and the antioxidative effect of Stachys japonica Miq (SJ) extract on FeCl2-induced cytotoxicity. For this study, superoxide anion-radical (SAR)-scavenging and superoxide dismutase (SOD)-like abilities with cell viability were done. FeCl2 showed a significant decrease of cell viability in dose-dependent manner, and it was mid-toxic. The caffeic acid showed a significant increase of cell viability against FeCl2-induced cytotoxicity. In the protective effect of SJ extract on FeCl2-induced cytotoxicity, it showed SAR-scavenging and SOD-like abilities with a significant increase of cell viability. From these results, the cytotoxicity of FeCl2 is correlated with oxidative stress, and SJ extract effectively protected the cytotoxicity of FeCl2 by antioxidative effect. Conclusively, the natural resources like SJ extract may be a useful fundamental materials for the development of an alternative antioxidant.

• Korean Chemical Engineering Research
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• v.57 no.6
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• pp.898-906
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• 2019

In this study, basic research was conducted to provide guidelines for selecting fluorescent dye and using proper concentration of fluorescent dye to use evaluation of cell viability and fluorescent dye delivery efficiency. Propidium iodide and Yo-Pro-1 were used as fluorescent dyes. In the evaluation of cell viability and the efficiency of delivery using Propidium Iodide and Yo-Pro-1, the histogram of each fluorescent dye was different depending on the type of fluorescent dye and the concentration used. These results were related to the characteristics of the fluorescent dyes used. This was related to the properties of the fluorescent dyes used. From these results, it was found that the analytical results depending on the characteristics of the fluorescent dyes used in the cell analysis. The effect of the fluorescent dye on the cell was confirmed, but it was confirmed that it did not affect the analysis result. In addition, the influence of interference between fluorescent signals was confirmed when two or more kinds of fluorescent dyes were used for analysis. The higher the concentration of Yo-Pro-1 was, the larger the effect of interference was, and the concentration of Propidium Iodide did not affect the interference of fluorescence signal. This study confirmed that the evaluation of the cell viability and the evaluation of the delivery efficiency were influenced by the type and concentration of the fluorescent dyes and it was related to the characteristics of the fluorescent dyes. Based on the results, appropriate concentrations of fluorescent dyes suitable for evaluation of cell viability and delivery efficiency were suggested.

• Radiation Oncology Journal
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• v.19 no.4
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• pp.381-388
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• 2001

Purpose : This study was conducted to assess the effects of x-irradiation on the expression of the novel glutathione S-transferase K1 gene. Materials and methods : Human glutathione S-transferase K1 (hGSTK1) DNA was purified and ligated to a pcDNA3.1/Myc-His(+) vector for the overexpression of hGSTK1 gene. MCF-7 cells were transfected with or without the recombinant hGSTK1 gene, and irradiated with 6 MV x-ray. After incubation of 14 days, cell survival was measured and compared. The expression of hGSTK1 and the effect of x-irradiation on hGSTK1 expression were also estimated in MCF-7 cells transfected with or without the hGSTK1 gene by RT-PCR. Results : Following 2 to 12 Gy of x-irradiation, the cell survivals were higher in the MCF-7 cells transfected with the hGSTK1 gene than in those without transfection. Despite the higher cell survival in the hGSTK1-transfected cells, RT-PCR for hGSTK1 mRNA revealed no significant differences according to radiation dose, fractionation, and time after irradiation. Conclusion : The MCF-7 cells transfected with the hGSTK1 gene showed higher cell survival than those without transfection of the gene. The hGSTK1 gene might be associated with the radiosensitivity of MCF-7 cell line and further analysis should be needed.

• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.385-393
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• 1997

다세포 생물에서 몸의 효율적 생존을 위한 각 기관의 homeostasis는 세포 증식과 사망에 의해 조절된다. 따라서, apoptosis라 명명된 세포사망은 정교한 기전에 의한 능동적이고 자발적인 사망기전으로써 몸의 정상적 유지를 위한 필수적인 현상이다. 발생기 세포나 신경세포 또는 흉선세포 분화 동안 과다한 세포의 제거가 apoptosis의 대표적인 예이며, 각종 호르몬에 의해 그 기능이 조절되는 난소세포에서도 apoptosis가 활발히 일어난다. Rat난소에는 태어날 때 수십 만개의 난포를 지니고 있는데, 이 중 단지 1%만이 배란에 사용되어질 뿐이고 나머지는 모두 사망하게 된다. 이러한 난포사망은 난소의 적절한 세포 수를 유지하기 위한 필수적 과정이며, 인위적으로 apoptosis를 억제하는 유전자인 bcl-2를 과다 발현시키면 난소암이 발생하는 연구결과가 이를 입증해주고 있다. 이처럼 중요한 난포 사망기전은 apoptosis라는 개념이 정립되면서 최근 들어 점차 그 연구가 활발해지고 있다. Apoptosis의 특징 중 뚜렷한 점은 DNA가 일정한 간격으로(180∼200 bp)잘려지는 DNA fragmentation현상으로, 이를 이용하여 DNA3'-end 부위에 방사선동위원소를 label한 후 이를 전기영동으로 분리하면 apoptosis를 손쉽게 측정할 수 있다. 난소의 기능은 시상하부호르몬인 LH와 FSH 뿐만 아니라 난소에서 분비되는 각종 난소국부호르몬들에 의해 조절된다. 특정한 발육단계의 난포는 특정한 호르몬에 의해 그 기능을 조절 받는데, 이러한 난소기능 조절기작은 매우 복잡한 경로를 지니고 있다. 이러한 복잡한 기작으로 인해 초기 연구에서첨 생체 내에서 밝히려는 연구 시도는 어려움에 부딪치게 되었다. 생체내 실험은 난소가 다양한 발육단계의 난포를 동시에 지니고 있어 특정한 발육단계의 난포 사망기전을 연구하기 어렵다. 또한 난포는 생체 내에서 다양한 호르몬을 동시에 분비하기 때문에 특정한 난소국부호르몬이 사망기전에 미치는 영향을 조사하기 힘든 점이 있다. 최근 들어 난포체외배양이 다양하게 개발되면서, 이러한 어려운 점을 극복할 수 있게 되었다. 본 논문은 각 발육단계의 난포를 절단해 체외배양하면서, apoptosis DNA 절단 현상을 이용하여 각종 난소국부 호르몬들이 난포발육단계별로 사망기전에 미치는 영향을 요약해 보였다. 난포는 발육하면서 점차 복잡한 호르몬 경로를 생존을 위해 필요로 한다. Prevulatory난포생존에 필요한 난소국부호르몬들은 early antral 단계의 난포에서는 그 미치는 영향이 감소되다가 preantral단계의 난포에서는 영향을 전혀 미치지 못했다. 단지 예외는 cGMP처리로써, 세포내 cGMP수준을 일정하게 유지시켜주는 것이 난포발육단계에 무관하게 생존에 중요한 인자로, 장래 연구는 난포 세포내의 cGMP수준을 조절하는 기작을 규명하는데 있을 것이다.

• Radiation Oncology Journal
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• v.19 no.2
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• pp.163-170
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• 2001

Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

• Tuberculosis and Respiratory Diseases
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• v.44 no.6
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• pp.1285-1295
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• 1997

Background : EGFR is one of the initial step in signal transduction pathway about multistep carcinogenesis. It is homologous to oncogene erbB-2 and is the receptor for EGF and TGF alpha. EGFR has important role in the growth and differentiation of tumor cells. So, EGFR in non-small cell lung cancer was examined to search for possible evidence as clinical prognostic factor. Methods : To investigate the role of EGFR in lung cancer, the author performed immunohistochemical stain of EGFR on 57 resected primary non-small cell lung cancer specimens. And the author analyzed the correlation between EGFR expression, clinical parameters, Sand $G_1$ phase fraction and survival. Results : 1) EGFR were detected in 56% of total 57 patients (according to histologic type, squamous cancer 50%, adenocarcinoma 63%, large cell cancer 75%) (according to TNM stage, stage I 64%, stage II 38%, stage III 55%) (according to cellular differentiation, well 50%, moderately 52%, poorly 65%). All differences were insignificant 2) Using the flow cytometric analysis, mean S-phase fraction of EGFR (+) and (-) group were 22.3(${\pm}10.5$)%. 18.0(${\pm}10.9$)% (p>0.05), mean $G_1$-phase fraction of EGFR (+) and (-) group were 68.4(${\pm}11.6$)%, 71.1(${\pm}12.8$)%, (p>0.05) 3) Two-year survival rate of EGFR (+) and (-) group were 53%, 84%, median survival time of EGFR (+) and (-) group were 26, 53 months. (p<0.05, Kaplan-Meier, generalized Wilcox) Conclusion : EGFR immunostaining may be a simple and useful method for survival prediction in non-small cell lung cancer.

• Journal of Chest Surgery
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• v.31 no.10
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• pp.973-981
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• 1998

Background: About 30% to 40% of the patients with pathologic stage I non-small cell lung cancer (NSCLC) die within 5 years after complete resection. The identification of poor prognostic factors and the application of additional treatment are very important to improve the survival rate in resected stage I NSCLC. Materials and methods: Sixty-eight(68) patients who had been diagnosed postoperatively between Janury 1989 and December 1995 as having stage I non-small cell lung cancer according to the TNM classification were studied. The postoperative 5-year survival rate was calculated with the Kaplan-Meier method, and clinico- histopathologic factors including age, sex, operative method, type of tumor cell, T factor, grade of the differentiation in a squamous cell carcinoma, invasion of blood vessel and expression of the nm23-H1 protein were investigated and analyzed. Results: The median survival of the entire group of patients was 58$\pm$3 months, with a 5-year survival of 58.9%. In univariate analysis, invasion of blood vessel and poor differentiation of the tumor cell in a squamous cell carcinoma significantly worsened the survival. In multivariate analysis, invasion of blood vessel and grade of the differentiation of the tumor cells in a squamous cell carcinoma remained independent prognostic factors. High expression of the nm23-H1 protein was related to a high postoperative 5-year survival in comparision with low expression of the nm23-H1 pretein (73.0% vs 50.7%), but there was no statistical significance. Conclusions: These results highlight the negative prognostic value of poor differentiation of tumor cells in a squamous cell carcinoma and invasion of blood vessel in stage I non-small cell lung cancer. Also, further studies are necessary to be determined prognostic value of the T factor and expression of the nm23 protein in non-small cell lung cancer.

• Korean Journal of Food Science and Technology
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• v.43 no.6
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• pp.787-790
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• 2011

The objective of this study was to investigate the anticancer effects of Acer tegmentosum M. extracts. HepG2 hepatocarcinoma cells were treated with ethanol, chloroform, ethylacetate, butanol, aqueous fraction and hot water extract. The antiproliferative effect was evaluated by trypan blue exclusion, MTT-based viability assay and morphology. The trypan blue test showed that anticancer effect of the A. tegmentosum M. extracts on HepG2 cells increased gradually in proportion to the increasing concentration of the fractions. The butanol fraction showed the highest anticancer activity against HepG2 cells (p<0.05). The MTT assay indicated that the growth inhibition by the butanol fraction was dose-dependent. These results suggest that A. tegmentosum M. has the potential to inhibit the growth of hepatocarcinoma cells.

• Journal of Dental Rehabilitation and Applied Science
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• v.36 no.3
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• pp.145-157
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• 2020

Purpose: This study was to determine the possible effects of trichloroacetic acid (TCA) and epidermal growth factor (EGF) through cell survival genes of the PI3K-AKT signaling pathway when applying an hydrophobically modified glycol chitosan (HGC)-based nanocontrolled release system to human gingival fibroblasts in oral soft tissue regeneration. Materials and Methods: An HGC-based nano-controlled release system was produced, followed by the loading of TCA and EGF. The group was divided into control (CON), TCA-loaded nano-controlled release system (EXP1), and the TCA- and EGF- individually loaded nano-controlled release system (EXP2). A total for 29 genes related to the PI3K-AKT signaling pathway were analyzed after 48h of culture in human gingival fibroblasts. Real-time PCR, 1- way ANOVA and multiple regression analysis were performed. Results: Cell survival genes were significantly upregulated in EXP1 and EXP2. From multiple regression analysis, ITGB1 was determined to be the most influential factor for AKT1 expression. Conclusion: The application of TCA and EGF through the HGC-based nano-controlled release system can up-regulate the cell survival pathway.

• Korean Journal of Food Science and Technology
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• v.34 no.3
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• pp.510-517
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• 2002

Effects of wheat arabinoxylan on mouse spleen lymphocytes and peritoneal macrophages were examined in vitro. Among three wheat arabinoxylans (A1: low MW, A2: medium MW, A3: high MW), A3$(50{\sim}100\;{\mu}g/mL)$ increased the viability of spleen lymphocytes up to $114{\sim}125%$ of the control. A1 and A3 $(20\;{\mu}g/mL)$ increased the viability of lipopolysaccharide-treated lymphocytes synergistically. Viability of murine peritoneal macrophages treated with wheat arabinoxylans $(10{\sim}100{\mu}g/mL)$ was increased up to $135{\sim}175%$ of the control. The cytotoxic activity of macrophages against murine lymphocytic leukemic cell increased in the presence of wheat arabinoxylan. Phagocytic index of macrophages treated with wheat arabinozylans $(20\;{\mu}g/mL)$ significantly increased $197{\sim}232%$ compared with the control, and lysosomal phosphatase and myeloperoxidase activities also increased significantly (p<0.05). Treatment of wheat arabinoxylans tended to decrease nitrite production, but significantly stimulated $H_2O_2\;and\;O_2$ productions of macrophages (p<0.05). These results indicate that the immunostimulating effect of wheat arabinoxylan may be closely related with lysosomal enzyme activity and reactive oxygen intermediate production of macrophages.