• Title/Summary/Keyword: A549 Cell

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UBE2S promotes the proliferation and survival of human lung adenocarcinoma cells

  • Liu, Zhi;Xu, Lijun
    • BMB Reports
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    • v.51 no.12
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    • pp.642-647
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    • 2018
  • Ubiquitin-conjugating enzyme E2S (UBE2S), a family of E2 protein in the ubiquitination process, is involved in development of various cancers. However, its role in lung adenocarcinoma, has not been well elucidated. In this report, we attempted to investigate expression and function of UBE2S in lung adenocarcinoma. Up-regulation of UBE2S at mRNA, and protein level, was observed in human cancer tissues and lung adenocarcinoma cells. Higher UBE2S expression correlated with poorer prognosis of lung adenocarcinoma patients. UBE2S expression was efficiently suppressed by lentivirus-mediated shRNA strategy in A549 cells, and UBE2S silencing led to reduced cell proliferation, colony formation, and enhanced apoptosis. Inverse results were observed, in UBE2S over-expressed H1299 cells. Microarray analysis indicated that a large number of genes were regulated by UBE2S, and p53 signaling pathway may be critical, to the role of UBE2S in cancer development. Together, UBE2S could be a potential target for lung adenocarcinoma.

Anti-cancer Potentials of Rhus verniciflua Stokes, Ulmus davidiana var. japonica Nakai and Arsenium Sublimatum in Human Gastric Cancer AGS Cells (AGS 인체위암세포에서 건칠, 유근피 및 신석 추출물의 항암 활성 비교 연구)

  • Baek, Ilsung;Im, Lyeng-Hae;Park, Cheol;Cho, Yung Hyun
    • Journal of Life Science
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    • v.25 no.8
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    • pp.849-860
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    • 2015
  • The anti-cancer activities of Rhus verniciflua Stokes (GC), Ulmus davidiana var. japonica Nakai (UGP) and arsenium sublimatum (SS) extracts, which have been used Oriental medicine therapy for various diseases, were investigated. The treatment of GC, UGP and SS alone, and combined treatment with GC, UGP and SS did not affect the cell viability in the mouse normal cell lines (RAW 264.7 macrophages and C2C12 myoblasts). However, co-treatment with GC, UGP and SS markedly induces apoptosis in human gastric cancer AGS cells, but not in other various cancer cell lines (human lung cancer A549, colon cancer HCT116, liver cancer Hep3B and bladder T24 cells) as evidenced by formation of apoptotic bodies, chromatin condensation, and accumulation of annexin-V positive cells. Co-treatment with GC, UGP and SS effectively induced the expression levels of Fas and Fas ligand, and inhibited the levels IAP family proteins such as XIAP, cIAP-1 and survivin, and anti-apoptotic Bcl-xL proteins compared with treatment with either agent alone. Combined treatment also significantly induced the loss of mitochondrial membrane potential, which was associated with the activation of caspases (-3, -8, and -9) and degradation of poly (ADP-ribose) polymerase. However, the cytotoxic effects induced by co-treatment with GC, UGP and SS were significantly attenuated by pan-caspases inhibitor, z-VAD-fmk, indicating an important role for caspases. These results indicated that the caspases were key regulators of apoptosis in response to co-treatment of GC, UGP and SS in human gastric cancer AGS cells and further studies will be needed to identify the active compounds.

Screening of Anticancer and Immune Activities by the Extracts of Fatsia japonica Decne. et Planch. (팔손이 용매별 추출물의 항암 및 면역 활성 탐색)

  • Kim, Dae-Ho;Kim, Jung-Hwa;You, Jin-Hyun;Kim, Cheol-Hee;Kwon, Min-Chul;Lee, Hak-Ju;Hwang, Baik;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.3
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    • pp.87-92
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    • 2005
  • The study was performed using of ethanol and water extracts of Fatsia japonica Decne. et Planch. in anticancer and immune activities. All extracts of 1.0 $g/{\ell}$ concentration were increased in over 60% of the anticancer activites in A549 and MCF7 cells. Root barks inhibited 55%, 74% in A549 and MCF7 cell by adding ethanol extracts of $g/{\ell}$ concentration. The cytotoxicity of human lung nomal cell (HEL299) counted up to about 22% for ethanol extracts of root barks in 1 $g/{\ell}$ concentration. The activity of human immune T and B cells were increased up to $140{\sim}170%$ by adding ethanol extract of the root barks. Increasing trend of secretion of cytokine (IL-6, $TNF-{\alpha}$) from human B and T cell for 5 days cultivation has been abserved. From the results, the anticancer and immune-stimulatory activities of the roots extract were higher than the extracts of other parts.

Antimutagenic and Cytotoxic Effects of Extracts of Kalopanax pictus NAKAI Endodermis (음나무 내피 추출물의 항돌연변이원성 및 세포독성 효과)

  • Kim, Myong-Jo;Kim, Ju-Sung;Kang, Won-Hee;Yeon, Kyu-Dong
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.2
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    • pp.132-138
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    • 2002
  • This study was performed to determine the antimutagenic and cytotoxic effect of ,Kalopanacis cortex endothelium. Methanol extract was used on Salmonella typhimurium TA98, TA100 and three cancer cell lines. In the Ames test, methanol extract of Kalopanacis cortex endothelium alone did not exhibit any mutagenicity but showed substantial inhibitory effects against mutation induced by 4-nitroquinoline-l-oxide(4NQO), N'-methyl- N'-nitro-N-nitrosoguanidine(MNNG) and benzo(a)pyrene(B(a)P). The methanol extract of Kalopanacis cortex endothelium showed approximately 79.29% and 75.38% inhibitory effect on the mutagenesis induced by 4NQO and B(a)P, respectively, against TA98 strain. Whereas 79.49%, 89.3% and 68.85% inhibitions were observed on the mutagenesis induced by 4NQO, MNNG and B(a)P against TA100 strain. Methanol extracts from Kalopanacis cortex endothelium showed high antimutagenic effects against 4NQO, MNNG and B(a)P. The anticancer effects of methanol extract from Kalopanacis cortex endothelium against human lung carcinoma(A549), human breast adenocarcinoma (MCF-7) and human hepatocellular carcinoma(Hep3B) were investigated. The treatment of 0.5mg/ml Kalopanacis cortex endothelium methanol extracts had the highest cytotoxicity against A549(81.54%), followed by MCF-7(81.92%) and Hep3B (78.57%). In contrast 0.5mg/ml treatment of methanol extracts from Kalopanacis cortex endothelium had only $4{\sim}25%$ cytotoxicity on normal human liver cell(293).

The Effects of Cement Alkalinity upon the Pore Water Alkalinity and the Chloride Threshold Level of Reinforcing Steel in Concrete

  • Nam Jingak;Hartt William H.;Kim Kijoon
    • Journal of the Korea Concrete Institute
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    • v.16 no.4 s.82
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    • pp.549-555
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    • 2004
  • Cement of three alkalinities (equivalent alkalinities of 0.36,0.52 and 0.97) was employed in fabricating a set of classical G109 type specimens. To-date, these have been subjected to a one week wet-one week dry cyclic pending using 15 w/o NaCl solution. At the end of the dry period, potential and macro-cell current were measured to indicate whether the top reinforcing steel was in the passive or active state. Once this bar became active, the specimen was autopsied and the extent of corrosion was documented. Subsequent to visual inspection, concrete powder samples were collected from the upper region of the top rebar trace; and at a certain times concrete cores were taken from non-reinforced specimens. Using these, determinations were made of (1) critical chloride concentration for corrosion initiation ($Cl_{th}^-$), (2) effective chloride diffusion coefficient ($D_e$), and (3) pore water alkalinity ($[OH^-]$). The pore water alkalinity was strongly related to the alkali content of cement that was used in the mix. The chloride concentration, ($Cl^-$), was greater at active than at passive sites, presumably as a consequence of electro migration and accumulation of these species at active site subsequent to corrosion initiation. Accordingly, ($Cl^-$) at passive sites was considered indicative of the threshold concentration fur corrosion initiation. The $Cl_{th}^-$ was increased with increasing Time-to-corrosion ($T_i$). Consequently, the HA(High Alkalinity) specimens exhibited the highest $Cl_{th}^-$ and the NA(Normal Alkalinity) was the least. This range exceeds what has previously been reported in North America. In addition, the effective diffusion coefficient, $D_e$, was about 40 percent lower for concrete prepared with the HA cement compared to the NA and LA(Low Alkalinity) ones.

Space grid analysis method in modelling shear lag of cable-stayed bridge with corrugated steel webs

  • Ma, Ye;Ni, Ying-Sheng;Xu, Dong;Li, Jin-Kai
    • Steel and Composite Structures
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    • v.24 no.5
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    • pp.549-559
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    • 2017
  • As few multi-tower single-box multi-cell cable-stayed bridges with corrugated steel webs have been built, analysis is mostly achieved by combining single-girder model, beam grillage model and solid model in support of the design. However, such analysis methods usually suffer from major limitations in terms of the engineering applications: single-girder model fails to account for spatial effect such as shear lag effect of the box girder and the relevant effective girder width and eccentric load coefficient; owing to the approximation in the principle equivalence, the plane grillage model cannot accurately capture shear stress distribution and local stress state in both top and bottom flange of composite box girder; and solid model is difficult to be practically combined with the overall calculation. The usual effective width method fails to provide a uniform and accurate "effective length" (and the codes fail to provide a unified design approach at those circumstance) considering different shear lag effects resulting from dead load, prestress and cable tension in the construction. Therefore, a novel spatial grid model has been developed to account for shear lag effect. The theoretical principle of the proposed spatial grid model has been elaborated along with the relevant illustrations of modeling parameters of composite box girder with corrugated steel webs. Then typical transverse and longitudinal shear lag coefficient distribution pattern at the side-span and mid-span key cross sections have been analyzed and summarized to provide reference for similar bridges. The effectiveness and accuracy of spatial grid analysis methods has been finally validated through a practical cable-stayed bridge.

Isolation of Anthraquinone Derivatives from the Root of Rumex japonicus H. (참소리쟁이 뿌리에서 안트라퀴논계 화합물의 분리 및 생리활성)

  • Hwang, Seon-Woo;Ha, Tae-Joung;Lee, Jong-Rok;Lee, Jun;Nam, Sang-Hae;Park, Ki-Hun;Yang, Min-Suk
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.274-278
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    • 2004
  • Four anthraquinone derivatives were isolated from the root of Rumex japonicus Houtt. These compounds were identified as physcion, emodin, chrysophanol-10,10'-bianthrone and $physcion-10,10'-bianthrone^(a)$, respectively. The last compound (a), especially, showed strong activity against A549, PC-3, UO-31 and HCT-15 human cancer cell lines with $IC_{50}$ values, ranging from 0.45 to $1.33\;{\mu}g/ml^{-1})$.

Ginsenoside Rf inhibits cyclooxygenase-2 induction via peroxisome proliferator-activated receptor gamma in A549 cells

  • Song, Heewon;Park, Joonwoo;Choi, KeunOh;Lee, Jeonggeun;Chen, Jie;Park, Hyun-Ju;Yu, Byeung-Il;Iida, Mitsuru;Rhyu, Mee-Ra;Lee, YoungJoo
    • Journal of Ginseng Research
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    • v.43 no.2
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    • pp.319-325
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    • 2019
  • Background: Ginsenoside Rf is a ginseng saponin found only in Panax ginseng that affects lipid metabolism. It also has neuroprotective and antiinflammatory properties. We previously showed that Korean Red Ginseng (KRG) inhibited the expression of cyclooxygenase-2 (COX-2) by hypoxia via peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$). The aim of the current study was to evaluate the possibility of ginsenoside Rf as an active ingredient of KRG in the inhibition of hypoxia-induced COX-2 via $PPAR{\gamma}$. Methods: The effects of ginsenoside Rf on the upregulation of COX-2 by hypoxia and its antimigration effects were evaluated in A549 cells. Docking of ginsenoside Rf was performed with the $PPAR{\gamma}$ structure using Surflex-Dock in Sybyl-X 2.1.1. Results: $PPAR{\gamma}$ protein levels and peroxisome proliferator response element promoter activities were promoted by ginsenoside Rf. Inhibition of COX-2 expression by ginsenoside Rf was blocked by the $PPAR{\gamma}-specific$ inhibitor, T0070907. The $PPAR{\gamma}$ inhibitor also blocked the ability of ginsenoside Rf to suppress cell migration under hypoxia. The docking simulation results indicate that ginsenoside Rf binds to the active site of $PPAR{\gamma}$. Conclusions: Our results demonstrate that ginsenoside Rf inhibits hypoxia induced-COX-2 expression and cellular migration, which are dependent on $PPAR{\gamma}$ activation. These results suggest that ginsenoside Rf has an antiinflammatory effect under hypoxic conditions. Moreover, docking analysis of ginsenoside Rf into the active site of $PPAR{\gamma}$ suggests that the compound binds to $PPAR{\gamma}$ in a position similar to that of known agonists.

Effects of Heat Shock Treatment on Enzymatic Proteolysis for LC-MS/MS Quantitative Proteome Analysis

  • Arul, Albert-Baskar;Han, Na-Young;Jang, Young-Su;Kim, Hyojin;Kim, Hwan-Mook;Lee, Hookeun
    • Mass Spectrometry Letters
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    • v.7 no.1
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    • pp.1-11
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    • 2016
  • Various efforts have been developed to improve sample preparation steps, which strongly depend on hands-on processes for accurate and sensitive quantitative proteome analysis. In this study, we carried out heating the sample prior to trypsin digestion using an instrument to improve the tryptic digestion process. The heat shock generated by the system efficiently denatured proteins in the sample and increased the reproducibility in quantitative proteomics based on peptide abundance measurements. To demonstrate the effectiveness of the protocol, three cell lines (A human lung cancer cell line (A549), a human embryonic kidney cell line (HEK293T), and a human colorectal cancer cell line (HCT-116)) were selected and the effect of heat shock was compared to that of normal tryptic digestion processes. The tryptic digests were desalted and analysed by LC-MS/MS, the results showed 57 and 36% increase in the number of identified unique peptides and proteins, respectively, than conventional digestion. Heat shock treated samples showed higher numbers of shorter peptides and peptides with low inter-sample variation among triplicate runs. Quantitative LC-MS/MS analysis of heat shock treated sample yielded peptides with smaller relative error percentage for the triplicate run when the peak areas were compared. Exposure of heat-shock to proteomic samples prior to proteolysis in conventional digestion process can increase the digestion efficiency of trypsin resulting in production of increased number of peptides eventually leading to higher proteome coverage.

Cytokines Stimulate Lung Epithelial Cells to Release Nitric Oxide

  • Robbins, Richard A.;Kwon, O-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.447-454
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    • 1995
  • Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

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