lung epithelial cell line a549barbour christina boots

Therefore, in this study we chose this experimental model to investigate the . MRC-5 and A549 cells were cultured in MEM and DMEM respectively, supplemented with 10% FBS, 1% sodium pyruvate, and 1% antibiotic-antimycotic solution. A549 cells were treated, after 12 h starvation, in 5% FBS medium according to scheme in Table 1. The results show that the A549 cell line is especially well suited for studying the control of expres-sion of xenobiotic-metabolizing CYPs in the human lung because this cell line expresses most of the major constitu-tive and inducible CYP forms found in lung epithelial cells. TMEM16A gene was transfected into A549 using Lipofectamine 2000. CompoZr Zinc Finger Nuclease (ZFN . A549 - adenocarcinoma based cell line of type II alveolar cells. 4. . RLE-6TN, a rat alveolar type II epithelial cell line, and A549, a human lung carcinoma epithelial cell line (ATCC, Manassas, VA, USA), were maintained in an F-12 medium in 75-cm 2 tissue culture flasks at 37.8C in 5% CO 2 and 95% air. A549; alveolar epithelial cells (Dulbecco's MEM + 10% FBS). Northern blot analysis demonstrated that steady-state IL-8 mRNA expression, by either TNF- or IL-1 beta-treated A549 cells, occurred in both a . The full HLA panel consists of a Beta-2 microglobulin (2M) knockout cell line and an additional ten (10) monoallelic HLA expression cell lines. The A549 (human lung adenocarcinoma), H1299 (human lung adenocarcinoma) cell lines were cultured with DMEM (Hyclone, Logan, UT, USA) containing 10% fetal bovine serum (FBS, Gibco BRL, Grand Island, NY, USA) at 37 C in a . . The A549 cell line is a well-characterized cellular model for asthma, allergies and respiratory infections. These cells grow adherently in monolayer and are suitable as a transfection host. Transfected cells were selected in the presence of G418 to create a stable TMEM16A overexpression A549 cell line. Our study reveals a previously unrecognized acute response of lung epithelial cells to cigarette . Importantly, the inhibition of Akt promoted GBS clearance both in alveolar epithelial cells in vitro and in lung tissue in vivo in a murine model of . Reverse transcriptase PCR HPL1 - SV40 transformed, normal human bronchial epithelial cells that are non tumorigenic. The A549 cell line was isolated in 1973 from a pulmonary adenocarcinoma1 in a study to . All lanes : Anti-MRP1 antibody at 1 g/ml Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate Lane 2 : ABCC1 knockout A549 (Human lung carcinoma cell line) whole cell lysate Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate Performed under reducing conditions. Background Cisplatin plus pemetrexed combination therapy is considered the standard treatment for patients with advanced, non-squamous, non-small-cell lung cancer (NSCLC). [1] HLC-1; well-differentiated bronchogenic adenocarcinoma (HamF12 + 10% FBS). In order to determine the function of YY1 in human lung epithelial cells, pSG5-YY1 plasmid was transfected into A549 cells. A549 cells form confluent monolayers with Type II characteristic morphology and tannic acid staining for typical lamellar bodies. Popular Answers (1) Interesting question - there are really no "normal" cell lines to compare with these cell types (A549 H1299), as all immortal cell lines mimic aspects of cancerous cells e.g . Confluent cells were exposed to the indicated concentrations of 17- -estradiol (E2) for 24 h followed by removal of medium, extraction of RNA, and real-time PCR experiments. Lanes 1 - 4: Merged signal (red and green). Ionizing radiation (IR) is used for patients diagnosed with unresectable non small cell lung cancer (NSCLC), however radiotherapy remains largely palliative due to radioresistance. The most sensitive peak for identifying dead cells is the 788 cm1peak corresponding to DNA O P O backbone stretching. The A549 cell line was used because it has been shown to produce IL-8 and many other mediators in response to lipopolysaccharide (LPS) (37. Four human lung cancer cell lines (Riken Cell Bank, Tokyo, Japan) were used for the experiments: 1. Cells between passages 5-8 were used for the experiments. As experimental model we used the human alveolar ther elucidate the complex interplay between metabolic pathways epithelial A549 cell line and selected two concentrations of Acrp30, and inammation, which occur at airway epithelium. A549 lung carcinoma. Prior to all experiments, cells were serum-starved for 18-24 h. (2) TMEM16A expression in A549 significantly increased at 24 hours and 36 hours, and then decreased at 48 hours after LPS treatment. A model for cytokine networks in the lung. One commonly used line is A549, an epithelial carcinoma derived from a 58 year old male patient, known to be KRAS mutant and EGFR wild type. A549 human lung cancer cells exhibited a pebble-like shape. A549 human lung epithelial cells were exposed to 0, 60, 140 and 200 g/cm2 doses of EHC-93 (total), its insoluble and soluble fractions for 24 h. Multiple cytotoxicity assays and proteomic analyses were used to assess particle toxicity in the exposed cells. 3 a. Abstract To investigate mechanisms regulating intra-alveolar coagulation, we studied monolayers of the A549 human lung epithelial cell line. The human lung epithelial cell line, A549, was purchased from the American Type Culture Collection (Manassas, VA) (19). Furthermore, volumes of various reagents are specific for the specified container size and must be adapted in each specific case. 340: 369-375, 1997. The surface of A549 cells delayed the onset of prothrombin-to-thrombin conversion and prevented total prothrombin consumption in normal plasma compared to plastic cell-free wells. To evaluate the expression of HSULF-1 in cells of pulmonary origin, five normal lung cells (fibroblasts (16Lu), fetal lung fibroblasts (HFL), primary lung fibroblasts (HLF), primary alveolar type 2 (hAT2) cells, and bronchial epithelial cells (HBE)) and five lung epithelial cancer cell lines (A549, H292, H1975 . These cells were chosen as they represent a model of human alveolar, type II pneumocytes for drug and toxin metabolism [52,53]. . The methods described below are designed for the A549 cell line from human lung epithelial cells. The A549 cells are characterized as a hypotriploid human alveolar basal epithelial cells and are widely used as an in vitro model for type II pulmonary epithelial cells as well as a model of lung adenocarcinoma [ 8 ]. The lung adenocarcinoma cell line, A549, undergoes epithelial-mesenchymal cell transition (EMT) in response to TGF-. IRE1-XBP1 regulates PDK1-dependent induction of epithelial-mesenchymal transition in non-small cell lung cancer cells . Proteomic profiling of the human alveolar derived cell line A549 revealed the most pronounced changes within the cellular secretome with preferential downregulation of proteins involved in wound healing and extracellular matrix organization. These cell lines present different EMT phenotypes as determined by the expression of EMT markers. A549 A549 A549D. A549-Dual cells are adherent epithelial cells that have been derived from the human A549 lung carcinoma cell line by stable integration of two inducible reporter constructs. However, advanced NSCLC has a 5-year survival rate of below 10%, which is mainly due to therapy resistance. Following implantation into immunocompromised mice, the cells form primary tumors and pulmonary metastases. In vitro cultured cells were treated with ILTPs; the proliferation of A549 cells and BEAS-2B human normal lung epithelial cells (Beas-2B cells) was observed using the 3- (4,5-dimethylazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the survival status of A549 cells was observed by fluorescence staining. this marker is very a549 lung epithelium cells in the rwv reactor provided op- densely and discretely expressed at junctional complexes in timal 3-d aggregate formation without compromising the via- 3-d cells since it is localized in or along the cell-cell contacts bility of the cells, and the 3-d aggregates grown for this period similar to Herein, we used two types of lung epithelial cells to compare animal and human alveolar performance. As control, cells were incubated with medium. After 16 hours, RFP, GFP, as well as non-transfected cells (A549) showed a significant increase in LDH release (RFP: 3.5 0.5-fold; A549: 3.3 1.1-fold; GFP: 4.1 1.0-fold) in response to high ZnO, whereas Luc showed only a slight increase (1.6 0.1-fold) that was not found to be statistically significant. Lung epithelial cells constitute the first defense line of host against the inhaled Aspergillus fumigatus; however, the transcriptional response of human alveolar type II epithelial cells was still unclear. Arch. A custom made unidirectional cell stretch device 23 was used to stretch A549 cells. Cell lines used in this study include A549, H1703, and H358 non-small cell lung cancer (NSCLC) cell lines ob-tained from the American Type Culture Collection. The dominant cell line used in this study was A549 human lung epithelial cells, a cell line derived from lung carcinoma. A549 CCL-185 A549 cells were isolated from the lung tissue of a White, 58-year-old male with lung cancer. These results establish that having retained several characteristics of human lung epithelial cell CYP expression, the A549 lung cell line is a valuable model for mechanistic studies on induction of the pulmonary CYP system. This is the first study to elucidate the mechanism through which the overexpression of ACE2 in the A549 lung cancer cell line decreases metastasis formation in vivo and upregulates the expression of E-cadherin both in vitro and in vivo. . Both CYPIA1 and CYPIIB6 P450 isozymes were determined to be functional with the fluorescent resorufin assay. Here, we studied the effects of proinflammatory cytokines derived from the mouse macrophage cell line RAW 264.7 on TGF--induced EMT in A549 lung cancer cells. Cells were washed, lysed, and sonicated to reduce DNA lengths to a range of 300-600 bp. The adenocarcinomic human alveolar basal epithelial cell line (A549) was maintained in RPMI 1640 medium (PAA Laboratories GMBH, Pasching, Austria) containing 10% FCS, 5% penicillin/streptomycin and 5% L-glutamine. This alveolar simple epithelial cell line forms a single layer in cell culture, provides an uncomplicated model for cell stretch with good microscopic access, and expresses keratin 8 (K8) and keratin18 (K18), two type I/II keratin isoforms with well . Cytogenetic Information The A549 cell line is hypotriploid with a modal chromosome number of 66, which occurs in 24% of cells. Product category Human cells Organism Homo sapiens, human Cell type epithelial cell Morphology epithelial-like Tissue Lung Disease Carcinoma Applications 3. A549: Lung carcinoma cell line: Giard et al., J Natl Cancer Inst 51: 1417 (1973) Lung: Male, 58: DSMZ: CVCL_0023 Cancer cell line: HBEC3-KT: Immortalized human bronchial epithelial cell line: Central lung bronchiole: Female, 65: Evercyte: CVCL_X491 Telomerase immortalized cell line: SCLC-21H: Small cell lung carcinoma cell line: Bepler et al . The A549 cell line is a widely used human lung adenocarcinoma cell line that was derived from a primary lung tumor. Images of individual wells were captured from bottom of the plate. TMEM16A mRNA and protein expressions in A549 were measured by real-time quantitative polymerase chain reaction (PCR) and Western blot, respectively. However, until now, the effect of quinolones on OPN expression in human lung epithelial cells remained to be elucidated. They were fixed with formaldehyde, then stained with 0.4% v / v crystal violet staining solution. We previously showed that the NSCLC cell line A549 harbors different subpopulations including a mesenchymal-like . H1703 has been classi-fied as mesenchymal, whereas A549 and H358 display a Lung epithelial cells constitute the first defense line of host against the inhaled Aspergillus fumigatus; however, the transcriptional response of human alveolar type II epithelial cells was still unclear. Here we used RNA-Seq technology to assess the transcriptome profiles of A549 cells following direct interaction with conidia of A. fumigatus. The elastic framework of the viabilityofBEAS-2Bcells,A549cells,andhumantrachealepithe- The human lung epithelial cell line A549 (CCL-185) and Calu-3 (HTB-55) were obtained from American Type Culture Collection and maintained as described by the supplier. Nicotine (N-3876) in liquid form was obtained from Millipore-Sigma (St. Louis, MO). 3 c). Induction of ferritin synthesis in human lung epithelial cells treated with crocidolite asbestos. Glucocorticoids do not prevent the EMT response, but TGF- induced resistance to the cytokine-regulatory action of glucocorticoids. Cell Line: A549: Species: Homo sapiens: Tissue: Lung: Morphology: Epithelial: Growth Properties: Adherent: Derivation: Derived from a 58 year old Caucasian male. Cancer stem cells (CSCs), as well as epithelial-mesenchymal transition (EMT), may contribute to drug and radiation resistance mechanisms in solid tumors. The RBP-J/DNA complexes were incubated with mouse antibody against RBP-J (Santa Cruz), or normal mouse IgG (Santa Cruz) for 18 hr at 4C. However, cancer cells have the plasticity and the heterogeneity even in the same cell line population, so that there are 2 possibilities. TRIP-1 regulates TGF-1-induced epithelial-mesenchymal transition of human lung epithelial cell line . After incubation period, proteins from cells as well as from human lung, porcine skeletal muscle and liver tissues were separated on SDS-PAGE as previously described ( Daniele et al., 2008 ). A549-Dual cells are adherent epithelial cells that have been derived from the human A549 lung carcinoma cell line by stable integration of two inducible reporter constructs. Here we used RNA-Seq technology to assess the transcriptome profiles of A549 cells following direct interaction with conidia of A. fumigatus. Elastase decreased the lung at the initial stage. Despite its function in tumor growth, the effects of H 2 S on ER stress in the human lung epithelial cell line A549 remain to be elucidated. The A549 cell line is a well-characterized cellular model for asthma, allergies and respiratory infections. 1) Some of A549 cells are epithelial, and others are . In the present study, hydrogen peroxide (H 2 O 2) was chosen as the model of lung injury in vitro. the lower, 5 g/ml, mimicking the human physiological serum lev- Chronic inammation has a crucial role in . The in vitro coculture system consisted of monolayers of human lung epithelial cell lines (A549 or NCI H441) and primary human pulmonary microvascular endothelial cells (HPMEC) on opposite sides of a permeable filter membrane. Cancer research, the generation of epithelial models of the distal lung for studies into airway function and disease and virology; the A549 cell line is a suitable host for many human The human tumorigenic lung epithelial cell line A549 was obtained from the American Type Culture Collection (CCL-185). Human primary bronchial epithelial cells respond differently to titanium dioxide nanoparticles than the lung epithelial . A549 cells were transfected with pN3-Sp1 or pN3 and then fixed with 1% polyformaldehyde for 10 min. 3-D A549 cell culture methods. mL 1 MCP1. A549 cells, grown in a 6-well plate, were incubated at 37 C for 2 weeks in the absence or presence of BP-LCNs (final concentration of 0.5, 1, 2.5, or 5 M). . 2. Lung epithelial A549 cell line was commercially provided by ATCC (#CCL-185; USA) and was cultured in F-12 K Medium (#30-2004; ATCC) supplemented with 10% FBS. Transfected the lung epithelial cells with lentivirus, then treated A549 cells with 100 M glycyrrhizin and treated BEAS-2B cells with 50 M glycyrrhizin for 24 h. (A) The relative HMGB1 mRNA expression was detected by RT-PCR to verify the transfection efficiency. Results: (1) Endogenous TMEM16A was expressed in rat AT-II and A549. The cells are adherent with an epithelial morphology. These cell lines present different levels of expression of epithelial-to-mesenchymal transition (EMT) markers such as E-cadherin, fibronectin, and vimentin. A human alveolar epithelial carcinoma cell line, A549 (CCL-185, American Type Culture Collection, Rockville, MD, USA) was cultured in RPMI medium 1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% heat-inactivated FBS (Gibco) at 37C and 5% CO 2. PubMed: 9143343 Geiger T, et al . Epithelial to mesenchymal transition (EMT), a complex process of cellular reprogramming has become an attractive drug target because it plays a crucial role in the metastasis of non-small cell lung cancer (NSCLC). HSULF-1 basal expression is lower in lung cancer cells than in normal lung cells. A549-Dual cells express a secreted embryonic alkaline . Here we investigated the molecular phenotype of A549 and H460 . A549 lung epithelial cells were treated with 1 ng/mL ricin alone or in combination with 100 ng/mL TRAIL, TNF-, or FasL for 4 h at 37 C followed by cell lysis and western blot. On the contrary, YY1 shRNA was used to knock down YY1 (Fig. Interleukin-8 gene expression by a pulmonary epithelial cell line. The overexpression of YY1 was confirmed as shown in Fig. 3 b). b) The proteasome inhibition showed a dose-dependent effect on IL8 secretion. J. Giard581972 [1] [2] A549 in vitro [1] [1] Biochem. . (3) TMEM16A mRNA and protein expressions were increased in the stable TMEM16A overexpression A549 cell line. ( A) The combination of ricin and TRAIL results in cleavage/activation of caspases-3, -7, -8, and -9. Treatment of the alveolar epithelial cell line A549 with the Akt inhibitor MK-2206 resulted in the enhanced production of reactive oxygen species and inflammatory mediators in response to GBS. Human ATII cells and HEK-293 T cells were purchased from Procell (#CP-H209 and #CL-0005; Wuhan, China) and cultured in human ATII cell complete culture medium and DMEM, respectively. Lee J.H. This cell line can be used in cancer, immuno-oncology, and toxicology research. The resulting T-CM was used as control medium in further analyses. A549, lung epithelial cell line was obtained from American Type Culture Collection (ATCC, Manassas, VA) and cultured in RPMI 10% FBS containing glutamine, penicillin, and streptomycin. It is known to have retained a number of the main characteristics of the native lung epithelium [13] such as tight junctions formation, presence of keratin, and synthesis of . The phenotypic change was accompanied by a progressive loss of epithelial markers and gain of mesenchymal markers. The human lung adenocarcinoma cell lines A549 and H1299 (American Type Culture Collection, Manassas, VA, USA) were cultured for 10 days: A549 in DMEM and H1299 in Roswell Park Memorial Institute 1640 medium (RPMI 1640) (Invitrogen), both supplemented with 10% FBS and 1% PS and grown at 37 C in 5% CO 2 and 95% relative humidity. lial cell line, A549 human type II lung cell line, and primary induces perivascular edema and an alveolar hemorrhage in cultures of human tracheal epithelial cells. Meanwhile, increased expression of -sma was observed following YY1 overexpression (Fig. LC-2/ad; moderately differentiated adenocarcinoma (HamF12 + RPMI1640 + 15% FBS + 25mM HEPES). . Co-culture and treatment with conditioned medium of RAW 264.7 cells enhanced a subset of TGF--induced EMT phenotypes in A549 cells, including changes in cell morphology and induction . The lung constitutes a primary site of exposure for many inhaled chemical toxicants and carcinogens. Using infection in human lung carcinoma cells we analyzed early and late host responses at 4, 8, 16 and 24 hours post infection by employing gene expression profiling on a microarray platform. Human lung carcinoma (A549) cells were . We investigated whether H 2 S protected A549 cells against oxidation-induced ER stress. A549 cells to be cultured in three dimensions were initially grown in Corning T75 cell culture asks containing GTSF-2 (HyClone) (22) supplemented with 10% was determined. The total number of identified genes was 19118 . This product is a panel of eleven (11) genetically modified cell lines targeting MHC Class I HLA molecules in an A549 lung carcinoma parental cell line background. The Raman spectrum of dead human lung derived (A549 line) cells indicates the breakdown of both phosphodiester bonds and DNA bases. : E64d; ; ALLN; : lactone. A549 cells were grown in 25 cm 2 tissue culture flasks (Corning Costar, Cambridge, MA) in Ham's F12 containing 10% fetal calf serum, Lglutamine (2 mM) and penicillinstreptomycin (100 U/ml100 mg/ml) until confluent . The amount of IL-6 and IL-1 proteins secreted into the culture medium was also increased after virus culture infection of A549 cell line compared to non-virus-cultured A549 cells and were significant in both IL-1 (p < 0.05 in 18 hours and p < 0.001 in 24-48 hours harvested supernatant) and IL-6 (p < 0.001). Materials and Methods Reagents 2,3,7,8 . When working with other cell lines, protocols may suffer some modification. Data were normalized to -actin expression. Cell line. The magnitude of this peak is reduced by 80% in the spectrum of dead cells. A549 cells are adenocarcinomic human alveolar basal epithelial cells, and constitute a cell line that was first developed in 1972 by D. J. Giard, et al. through the removal and culturing of cancerous lung tissue in the explanted tumor of a 58-year-old caucasian male. Biophys. to induce gene expression of IL-8 from pulmonary type II-like epithelial cells (A549). We sought to characterize the impairment of glucocorticoid response in A549 cells. To determine if our results were unique to A549 cells, we conducted experiments with Calu3 human . A recent report demonstrated that a human lung type II epithelial cell line (A549) is a suitable model to study host defense cellular responses . Bucht A. A549-Dual cells express a secreted . . . So they are different both in terms of. The A549 cell line (American Type Culture Collection - CCL-185; human, epithelial . The A549 cell line was derived from human lung carcinoma of a 58-year-old Caucasian male [11] which has been routinely used as a model for lung epithelial cells [12]. To elucidate this mechanism, we have silenced hnRNPA2/B1 mRNA in non-small-cell lung cancer cell lines A549, H1703, and H358. Human lung alveolar epithelial carcinoma A549 cells (CCL-185, ATCC, Manassas, VA) were grown in F12-K culture medium supplemented with 10% fetal bovine serum (ATCC) and subcultured at 80-90% confluency. Lung cancer is the leading cause of cancer-related deaths in the world with non-small cell lung cancer (NSCLC) making up about 85% of all lung cancer cases. The in vitro coculture system consisted of monolayers of human lung epithelial cell lines (A549 or NCI H441) and primary human pulmonary microvascular endothelial cells (HPMEC) on opposite sides of. A549 ( ATCC CCL-185 ) is a human lung adenocarcinoma cell line initiated through explant culture of lung carcinomatous tissue from a 58-year-old Caucasian male. Human lung alveolar epithelial cell line A549 was obtained from the American Type Culture Collection (Manassas, VA, United States). In the present study, we examined the effects of withaferin A (WFA), a plant-derived steroidal lactone on EMT in human NSCLC cell lines.