• Title/Summary/Keyword: Cell-based assay

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MicroRNA-23a: A Novel Serum Based Diagnostic Biomarker for Lung Adenocarcinoma

  • Lee, Yu-Mi;Cho, Hyun-Jung;Lee, Soo-Young;Yun, Seong-Cheol;Kim, Ji-Hye;Lee, Shin-Yup;Kwon, Sun-Jung;Choi, Eu-Gene;Na, Moon-Jun;Kang, Jae-Ku;Son, Ji-Woong
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.1
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    • pp.8-14
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    • 2011
  • Background: MicroRNAs (miRNAs) have demonstrated their potential as biomarkers for lung cancer diagnosis. In recent years, miRNAs have been found in body fluids such as serum, plasma, urine and saliva. Circulating miRNAs are highly stable and resistant to RNase activity along with, extreme pH and temperatures in serum and plasma. In this study, we investigated serum miRNA profiles that can be used as a diagnostic biomarker of non-small cell lung cancer (NSCLC). Methods: We compared the expression profile of miRNAs in the plasma of patients diagnosed with lung cancer using an miRNA microarray. The data from this assay were validated by quantitative real-time PCR (qRT-PCR). Results: Six miRNAs were overexpressed and three miRNAs were underexpressed in both tissue and serum from squamous cell carcinoma (SCC) patients. Sixteen miRNAs were overexpressed and twenty two miRNAs were underexpressed in both tissue and serum from adenocarcinoma (AC) patients. Of the four miRNAs chosen for qRT-PCR analysis, the expression of miR-23a was consistent with microarray results from AC patients. Receiver operating characteristic (ROC) curve analyses were done and revealed that the level of serum miR-23a was a potential marker for discriminating AC patients from chronic obstructive pulmonary disease (COPD) patients. Conclusion: Although a small number of patients were examined, the results from our study suggest that serum miR-23a can be used in the diagnosis of AC.

Biological Activity and Biochemical Properties of Water Extracts from Bacillus subtilis-fermented Silkworm (Bombyx mori L.) Powder by Origin (산지별 고초균 발효누에의 이화학적 특성 및 생리활성)

  • Kim, Tae-Hoon;Ahn, Hee-Young;Kim, Young-Wan;Sim, So-Yeon;Seo, Kwon-Il;Cho, Young-Su
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1470-1478
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    • 2017
  • The aim of this study was to investigate biological activity and biochemical properties of extracts from Bacillus subtilis-fermented silkworm (Bombyx mori L., SP) powder of different origin (Buan, Namwon, and Boeun). An additional aim was to determine the inhibition of cancer cell (B16-F10, HT-29, LNcaP, and MCF-7) proliferation and nitric oxide (NO) production from lipopolysaccharide (LPS)-induced RAW264.7 cells. Biological activities (${\alpha},{\alpha}^{\prime}$-diphenyl-${\beta}$-picrylhydrazyl [DPPH], free radical scavenging activity, fibrinolytic activity, antiproliferation activity, and anti-inflammatory activity) and biochemical properties (compositional amino acid contents, and mineral contents) were examined in water extracts from silkworm powder and B. subtilis-fermented silkworm powder. The highest amino acid contents were detected in Buan silkworm powder (BU). After fermented, the highest contents were found in B. subtilis-fermented Buan silkworm powder (BBO). The major minerals detected were K, Ca, and Mg. Rates of these minerals, especially those of Na increased after fermented. DPPH radical scavenging activity and fibrinolytic activity were stronger in the fermented group than non-fermented group. DPPH radical scavenging activity and fibrinolytic activity were highest in the extract from BBO. The inhibition activities of LNcaP and MCF-7 cells viability were significantly decreased in the BBO, and there was no inhibition activity in other cancer cells (B16-F10 and HT-29). An SRB assay of the cell viability of RAW 264.7 cells exposed to extracts of silkworm powder and B. subtilis-fermented silkworm powder revealed no toxicity in any of the groups. Compared with the LPS-treated group, the biggest reduction in NO production was detected in the BBO group. Based on these results, extracts from Boeun silkworm powder fermented with B. subtilis could be a candidate material as a dietary supplement for use in healthy functional foods.

Skin Moisturizing Properties and Anti-Inflammatory effects of extracts from Coptis chinensis in HaCaT cells (HaCaT cell에서 황련 추출물(Coptis chinensis)의 피부보습과 항염증 효과)

  • Kim, Eun-Hee;Moon, Young-Lan;Jang, Young-Ah
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.3
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    • pp.870-882
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    • 2021
  • Coptis chinensis has been used in the treatment of various diseases such as soothing, anti-inflammation, antimicrobial and antipyretic in oriental traditional medicine. In this study, we investigated the effect of hot water extract of Coptis chinensis(CCW) on skin barrier and inflammation-related factors in UVB and TNF-α/IFN-γ-induced HaCaT cells and evaluated its potential as a moisturizing and anti-inflammatory material. Based on result, the amount of HA (Hyaluronic acid) production and protein and mRNA expression of filaggrin were measured. In TNF-α/IFN-γ-induced HaCaT cells, CCW increased the amount of HA production in a concentration-dependent manner. In the measurement of protein and mRNA expression of filaggrin, the expression rate increased as the concentration of CCW increased. In UVB-induced HaCaT cells, CCW decreased the production of ROS and showed significant results with EGCG ((-)-epigallocatechin-3-gallate), a positive control. In addition, CCW inhibited the expression of inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-8 in TNF-α/IFN-γ-induced HaCaT cells. It was confirmed that the protein and mRNA expression of COX-2, a major factor in skin inflammation, was decreased in a concentration-dependent manner. These results suggest that hot water extract from Coptis chinensis can be used as a cosmetic material having a moisturizing and anti-inflammatory effect.

Characterization of ginsenoside compound K loaded ionically cross-linked carboxymethyl chitosan-calcium nanoparticles and its cytotoxic potential against prostate cancer cells

  • Zhang, Jianmei;Zhou, Jinyi;Yuan, Qiaoyun;Zhan, Changyi;Shang, Zhi;Gu, Qian;Zhang, Ji;Fu, Guangbo;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.228-235
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    • 2021
  • Backgroud: Ginsenoside compound K (GK) is a major metabolite of protopanaxadiol-type ginsenosides and has remarkable anticancer activities in vitro and in vivo. This work used an ionic cross-linking method to entrap GK within O-carboxymethyl chitosan (OCMC) nanoparticles (Nps) to form GK-loaded OCMC Nps (GK-OCMC Nps), which enhance the aqueous solubility and stability of GK. Methods: The GK-OCMC Nps were characterized using several physicochemical techniques, including x-ray diffraction, transmission electron microscopy, zeta potential analysis, and particle size analysis via dynamic light scattering. GK was released from GK-OCMC Nps and was conducted using the dialysis bag diffusion method. The effects of GK and GK-OCMC Nps on PC3 cell viability were measured by using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Fluorescent technology based on Cy5.5-labeled probes was used to explore the cellular uptake of GK-OCMC Nps. Results: The GK-OCMC NPs had a suitable particle size and zeta potential; they were spherical with good dispersion. In vitro drug release from GK-OCMC NPs was pH dependent. Moreover, the in vitro cytotoxicity study and cellular uptake assays indicated that the GK-OCMC Nps significantly enhanced the cytotoxicity and cellular uptake of GK toward the PC3 cells. GK-OCMC Nps also significantly promoted the activities of both caspase-3 and caspase-9. Conclusion: GK-OCMC Nps are potential nanocarriers for delivering hydrophobic drugs, thereby enhancing water solubility and permeability and improving the antiproliferative effects of GK.

Antioxidant, Anti-inflammatory and Anti-photoaging Activities of Hydrolyzed Peony (Paeonia lactiflora Pall.) Flower (작약꽃 가수분해물의 항산화, 항염 및 광노화 억제 효능)

  • Kyung Ju Lee;You Ah Kim;Byoung Jun Park
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.259-267
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    • 2023
  • This study was conducted to evaluate physiological activity of flower extract of peony (Paeonia lactiflora Pall.) by hydrolysis and to use it as a valuable cosmetic ingredients. Four cultivar of peony flowers were extracted, and the highest active ingredient contents was selected, and that cultivar was used for hydrolyzing. The results showed that high concentration of hydrochloric acid (HCl) hydrolyzed, and biological hydrolysis using enzymes had no activity. The deglycosylation of peonidin 3,5-diglucoside occurred by hydrolysis. The hydrolysate contains 63.3 ppm of peonidin, a red-colored anthocyanin compound. The antioxidant activity of hydrolysate was compared with extract. The results showed the strong antioxidant activity in hydrolysate (96%) than extract (82%). In addition, hydrolysate of peony flower showed higher inhibitory activity of NO release than extract. UVA assay using fibroblast cell (CCD-986Sk) showed that hydrolysate increased cell viability than extract under UVA exposure. Based on these results, we anticipate that hydrolysate of peony flower can be used a valuable cosmetic ingredient.

Anti-proliferative Activities of Solvent Fractions of Lees Extracts in Human Colorectal HCT116 Cells (대장암 세포주에서 주박 추출물의 유기용매 분획물의 항성장 활성)

  • Kang, Hyung-Taek;Lee, Seung Hoon;Kim, Soon Young;Kim, Mi-Sun;Shin, Woo-Chang;Sohn, Ho-Yong;Kim, Jong-Sik
    • Journal of Life Science
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    • v.24 no.9
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    • pp.967-972
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    • 2014
  • In the present study, we prepared eighty-five different kinds of lees extracts and their solvent fractions and investigated their anti-proliferative activities against human colorectal cancer HCT116 cells. HCT116 cells were treated with eighty-five solvent fractions of lees extracts and then cell viability was measured using MTS assay. Among the treated solvent fractions, three solvent fractions (KSD-E1-3, KSD-E2-3, and KSD-E4-3) were selected based on cell viability assay. In addition, we performed an oligo DNA microarray analysis to analyze the gene expression changes by treatment of KSD-E1-3 in HCT116 cells. Among the upregulated genes, we selected 4 genes (NAG-1, ATF3, p21, and DDIT3) and performed RT-PCR using gene-specific primers. Among the treated solvent fractions, KSD-E1-3 dramatically induced the expressions of the four selected genes. In addition, we investigated whether the upregulations of those genes were dependent on the transcription factor p53's presence using p53 null HCT116 cells. The results indicate that the upregulations of NAG-1, ATF3, and DDIT3 are not dependent on the p53 presence, whereas p21 is dependent on the p53 presence. These findings may help to understand the molecular mechanisms of the anti-proliferative activity mediated by rice wine lees in human colorectal cancer cells.

Suppressive Effects of Epigallocatechin Gallate Pretreatment on the Expression of Inflammatory Cytokines in RAW264.7 Cells Activated by Lipopolysaccharide (Lipopolysaccharide로 활성화된 RAW264.7세포에서 염증반응사이토카인 발현에 대한 Epigallocatechin gallate의 억제효능연구)

  • Seo, Eun Ji;Go, Jun;Kim, Ji Eun;Koh, Eun Kyoung;Song, Sung Hwa;Sung, Ji Eun;Park, Chan Kyu;Lee, Hyun Ah;Kim, Dong Seob;Son, Hong Joo;Lee, Cung Yeoul;Lee, Hee Seob;Hwang, Dae Youn
    • Journal of Life Science
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    • v.25 no.9
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    • pp.961-969
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    • 2015
  • Epigallocatechin gallate (EGCG), the main catechin in green tea, has been shown to have some beneficial effects against various human diseases, including diabetes, neurodegenerative disorders, cancer, cardiovascular disease and obesity. To investigate the mechanism of the suppressive effects of EGCG on inflammatory response in macrophages, alterations on the levels of nitric oxide (NO) regulatory factors and inflammatory cytokines were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells. No significant toxicity was detected in RAW264.7 cells treated with 100–400 μM EGCG. Moreover, the optimal concentration of LPS was determined to be 1 μg/ml based on the results of cell viability assay, NO assay and IL-6 enzyme-linked immunsorbent assay (ELISA). Furthermore, NO levels decreased significantly by 68.2% in the 400 μM EGCG/LPS treated group, while the level of inducible nitric oxide synthase (iNOS) expression decreased by 12-17% in the 200 and 400 μM EGCG/LPS treated group. A significant decrease in transcription of pro-inflammatory cytokines (TNF- α and IL-1β) and anti-inflammatory cytokine (IL-10) was also detected in the EGCG/LPS treated group. However, IL-6 transcript and protein was maintained at a constant level when in the LPS treated group relative to the EGCG/LPS treated group. Overall, these results suggest that the differential regulation of inflammatory cytokines is an important factor influencing the suppressive effects of EGCG against LPS-activated inflammatory response in RAW264.7 cells.

Anti-Inflammatory Effects of Picrasma Quassioides (D.DON) BENN Leaves Extracts (소태나무 잎 추출물의 항염증 효과)

  • Jung, Yeon Seop;Eun, Cheong Su;Jung, Young Tae;Kim, Hyun Jeong;Yu, Mi Hee
    • Journal of Life Science
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    • v.23 no.5
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    • pp.629-636
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    • 2013
  • This study was performed to evaluate the anti-inflammatory and antioxidant activities of methanol extract from the leaves of Picrasma quassioides BENNET (PLME). The antioxidant effects of PLME were measured based on polyphenol and flavonoid contents. PLME was found to have $367.52{\mu}g/mg$ and $46.61{\mu}g/mg$ high polyphenol and flavonoid contents. Cell viability was determined by MTT assay. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was measured by Griess assay and enzyme-linked immunosorbent assay (ELISA). In order to effectively anti-inflammatory agents, we examined the inhibitory effects on the production of lipopolysaccharide (LPS)-induced NO and $PGE_2$ in RAW 264.7 cells. PLME significantly decreased the production of NO and $PGE_2$ in a dose-dependent manner, and also reduced the expression of iNOS, a COX-2 protein. In addition, PLME reduced the NF-${\kappa}B$, $I{\kappa}B$ phosphorylation in RAW 264.7 cells upon stimulation with LPS (100 ng/ml) for 24 h. These results provide evidence for the anti-inflammatory and antioxidant effects of Picrasma quassioides leaves.

Development of Human Antibody Inhibiting RNase H Activity of Polymerase of Hepatitis B Virus Using Phage Display Technique (Phage Display 기법을 이용한 B형 간염 바이러스 Polymerase의 RNase H 활성을 억제하는 인간 단세포군 항체의 개발)

  • Lee, Seong-Rak;Song, Eun-Kyoung;Jeong, Young-Joo;Lee Young-Yi;Kim, Ik-Jung;Choi, In-Hak;Park, Sae-Gwang
    • IMMUNE NETWORK
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    • v.4 no.1
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    • pp.16-22
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    • 2004
  • Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.

Antitumor Activity and Nephrotoxicity of the Novel Platinum(II) Coordination Complex (새로운 Platinum (II) Complex [Pt (II)(trans-d-dach)(DPPE)] $(NO_3)_2$의 항암효과 및 신독성에 관한 연구)

  • Jung Jee-Chang;Lee Moon-Ho;Chang Sung-Goo;Rho Young-Soo
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.103-114
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    • 1995
  • Platinum coordination complexes are currently one of the most compounds used in the treatment of solid tumors. However, its use is limited by severe side effects such as renal toxicity. Our platinum-based drug discovery program is aimed at developing drugs capable of diminishing toxicity and improving antitumor activity. We synthesized new Pt (Ⅱ) complex analogue containing 1,2-diaminocyclohexane (dach) as carrier ligand and 1,2-bis(diphenylphosphino) ethane (DPPE) as a leaving group. Furthermore, nitrate was added to improve the solubility. A new series of [Pt(trans-ddach)(DPPE).$2NO_3(PC)$ was synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared (IR), $^{13}carbon$ nuclear magnetic resonance (NMR)]. PC demonstrated acceptable antitumor activity aganist P388, L-1210 lymphocytic leukemia cells and SK=OV3 human ovarian adenocarcinoma cells, and significant. activity as compared with that. cisplatin. The toxicity of PC was found quite less than thar of cisplatin using MTT, $[^3H]$ thymidine uptake and glucose consumption tests in rabbit proximal tubule cells, human kidney cortical cells and human renal cortical tissues. Based on these results, this novel platinum compound represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low toxicity.

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