non small cell lung cancer nsclc cell lines a549  (ATCC)

Journal: Science Advances

Article Title: Epithelial plasticity shapes intratumoral heterogeneity and cell lineages in early-stage lung cancer

doi: 10.1126/sciadv.ady8546

Figure Lengend Snippet: ( A ) Box plots showing KLF4 mRNA expression in the A549 LUAD cell line in the overexpression group and the control group. The median and interquartile range are shown for each cluster, two-sided Wilcoxon test. ( B ) Cell viability assay showing that overexpression of the KLF4 gene in KLF4 low LUAD A549 cells decreases cell proliferation. ( C ) Colony formation assays confirming that overexpression of the KLF4 gene decreases cancer cell proliferation in the A549 cells. ( D ) Transwell assays confirmed that overexpression of the KLF4 gene decreases cell migration in the A549 cells. ( E ) Soft agar assay showing that overexpression of the KLF4 gene inhibits anchorage-independent growth in the A549 cells. ( F and G ) Growth curves of xenograft tumors (F) and subcutaneous tumor size (G) of each group showing that the overexpression of the KLF4 gene inhibits tumor growth. ( H ) CUT&Tag-seq tracks of the gene of transitional marker and C2 cluster–specific genes locus in the indicated cells. KLF4-OE: Flag-KLF4–overexpressing A549 cells (anti-Flag antibody); Ctrl: Flag-overexpressing A549 cells (anti-Flag antibody). ( I ) Volcano plot showing the transitional marker and C2 cluster–specific genes distribution with respect to KLF4 expression levels. ( J and K ) GSEA showing that the KLF4 + cancer cell (J), AT1 cell, and AT2 cell (K) features were affected by high expression of KLF4.

Article Snippet: Human embryonic kidney (HEK) 293T cells [American Type Culture Collection (ATCC), catalog #ACS-4500) and the non–small cell lung cancer (NSCLC) cell lines A549 (ATCC, catalog #CCL-185) and NCI-H1437 (ATCC, catalog #CRL-5872) were used.

Techniques: Expressing, Over Expression, Control, Viability Assay, Migration, Soft Agar Assay, Marker

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Clonogenic Survival of Lung Carcinoma Cells. Doxorubicin- and naphthalimide-induced inhibition of clonogenic survival. ( a ) Representative images of A549 and H1299 cell colonies captured on 10–14 days post-treatment with doxorubicin, mitonafide, and 5a at concentrations corresponding to IC 10 , IC 15 , and IC 20 . Untreated controls formed numerous dense colonies, whereas treatment markedly reduced both colony number and size in a concentration-dependent manner; ( b ) Quantitative analysis of clonogenic survival of A549 and H1299, expressed as surviving fraction relative to untreated controls. Data represent the mean values of at least three independent experiments.

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Inhibition, Concentration Assay

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Effect of Compound 5a on Cell Cycle Distribution in A549 Cells. Flow-cytometric analysis of propidium iodide (PI)-stained cells after 24 h and 48 h treatment with 5a at concentrations corresponding to IC 50 and IC 75 . ( a , c ) Representative histograms of PI fluorescence profiles for 24 h and 48 h treatment, respectively (black lines—row data; green lines—fitted model lines generated by the Dean-Jett-Fox algorithm). ( b , d ) Quantification of cell-cycle phase distribution (G 1 , S, and G 2 /M) expressed as a percentage of the total cell population. For each sample, at least 100,000 events were acquired. Data represent mean ± SD of three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test ( p < 0.05). Statistical significance was assessed within each cell-cycle phase to compare doses independently of other phases. Groups sharing at least one identical letter are not significantly different from each other ( p > 0.05), while groups with different letters differ significantly ( p < 0.05).

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Staining, Fluorescence, Generated

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Replication Stress and DNA Damage Induced by Compound 5a in A549 and H1299 cells. Immunofluorescence analysis of EdU incorporation and γH2AX foci formation in A549 ( a , b ) and H1299 ( c , d ) cells following treatment with compound 5a at IC 50 and IC 75 for 24 h and 48 h. Scale bar: 20 µm. Images were acquired using identical exposure and analysis settings across all conditions; quantification was performed using uniform thresholding criteria. Quantitative analysis was performed on at least 10 randomly selected fields per condition per experiment, from three independent experiments. Mean fluorescence intensity corresponds to the ratio of the measured signal intensity to the area of the cell nucleus. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. Groups sharing at least one identical letter are not significantly different from each other ( p > 0.05), while groups with different letters differ significantly ( p < 0.05).

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Immunofluorescence, Fluorescence

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Annexin V–FITC/PI Apoptosis Analysis. ( a , c ) Representative Annexin V-FITC/PI dot-plots of A549 and H1299 cells treated with IC 50 concentrations of 5a for 24 h and 48 h. ( b , d ) Quantitative distribution, in percentage, of viable (V—light green), early apoptotic (EA—green), late apoptotic (LA—orange), and necrotic (N—red) populations in A549 and H1299 cells. At least 20,000 events per sample were acquired. Bars represent mean percentages of cells. Error bars indicate SD ( n = 3 independent experiments) and are shown in the positive direction only for clarity in stacked bar plots.

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Caspase-3/7 Activation Induced by Compound 5a . ( a , c ) Representative immunofluorescence images of A549 and H1299 cells after 24 h treatment with 5a at concentrations corresponding to IC 50 and IC 75 . Activated caspase 3/7 is visualized as green fluorescence. Nuclei are counterstained with DAPI (blue). Scale bar: 20 µm; ( b , d ) Quantitative analysis of caspase 3/7 activation based on mean fluorescence intensity from at least 10 fields per condition per experiment from three independent experiments. Mean fluorescence intensity corresponds to the ratio of the measured signal intensity to the area of the cell nucleus. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. Groups sharing at least one identical letter are not significantly different from each other ( p > 0.05), while groups with different letters differ significantly ( p < 0.05).

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activation Assay, Immunofluorescence, Fluorescence

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Nuclear Accumulation of p53 in A549 Cells. ( a ) Representative immunofluorescence images of control and treated A549 cells after a 24 h exposure to IC 50 and IC 75 concentrations of 5a . Cells were labeled with anti-p53 antibody (red) and counterstained with DAPI (blue). Scale bar: 20 µm; ( b ) Quantitative determination of mean p53 fluorescence intensity. Mean fluorescence intensity corresponds to the ratio of the measured signal intensity to the area of the cell nucleus. At least 10 randomly selected fields per condition were scored, from three independent experiments. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. Groups sharing at least one identical letter are not significantly different from each other ( p > 0.05), while groups with different letters differ significantly ( p < 0.05).

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Immunofluorescence, Control, Labeling, Fluorescence

Journal: Pharmaceutics

Article Title: Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

doi: 10.3390/pharmaceutics18020167

Figure Lengend Snippet: Schematic Representation Summarizing the Proposed Mechanism of Action of Compound 5a in Lung Carcinoma Cells. Compound 5a induces pronounced replication stress, as evidenced by inhibition of DNA synthesis (EdU suppression) and accumulation of DNA damage (γH2AX). In p53-proficient A549 cells, this response is associated with nuclear accumulation of p53 and G 1 -phase cell cycle arrest, whereas in p53-deficient H1299 cells, checkpoint activation predominantly manifests as G 2 /M arrest. Sustained DNA damage subsequently triggers caspase-dependent apoptosis, as confirmed by Annexin V/PI staining and executioner caspase-3/7 activation. The schematic summarizes the convergence of replication stress, checkpoint engagement, and apoptotic cell death in both p53-dependent and p53-independent cellular contexts.

Article Snippet: The human non-small cell lung cancer cell lines A549 (ATCC ® CCL-185TM) and NCI-H1299 (ATCC ® CRL-5803TM), along with the normal lung fibroblast line MRC-5 (ATCC ® CCL-171TM), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Inhibition, DNA Synthesis, Activation Assay, Staining

Journal: Journal of Translational Medicine

Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment

doi: 10.1186/s12967-025-07450-x

Figure Lengend Snippet: Effects of Tr-ACT2 on a lung cancer cell model. ( A ) Effects of Tr-ACT2 on the growth of A549 cells at concentrations of 0.05 µM, 0.25 µM, and 1 µM for indicated time. Cell growth was evaluated using the MTT assay. ( B ) Representative images of A549 cell morphology at 48 h after treatment with 0.25 μM Tr-ACT2 or vehicle. Scale bar: 400 μm. ( C , D ) Western blot analysis showing the protein levels of PARP, cleaved PARP, IgG heavy chain, IgG light chain, and GAPDH in A549 cells treated with 0.25 μM Tr-ACT2 for indicated time. Protein levels are quantified by densitometry and normalized by GAPDH, versus 0 h. Data are presented as means ± SD ( n ≥ 4). p > 0.05 (ns), p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****)

Article Snippet: Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human non-small cell lung cancer (NSCLC) cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37 °C and 5% CO2.

Techniques: MTT Assay, Western Blot

Journal: Journal of Translational Medicine

Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment

doi: 10.1186/s12967-025-07450-x

Figure Lengend Snippet: Antitumor efficacy of Tr-ACT2 in the A549 xenograft model. ( A ) Tumor volume change over time for all mice. ( B ) Normalized body weight change over time for all mice. Data are presented as means ± SEM ( n ≥ 3)

Article Snippet: Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human non-small cell lung cancer (NSCLC) cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37 °C and 5% CO2.

Techniques: