Journal: Oncology Letters

Article Title: Effects of tanshinones mediated by forkhead box O3a transcription factor on the proliferation and apoptosis of lung cancer cells

doi: 10.3892/ol.2018.9530

Figure Lengend Snippet: Representative images of transfected A549 cells (magnification, ×200). (A) A549 cells were transfected with siRNA1. (B) A549 cells were transfected with siRNA2. (C) A549 cells were transfected with siRNA3. (D) A549 cells were transfected with ssiRNA. siRNA, small interfering RNA; ssiRNA, scrambled siRNA.

Article Snippet: The human lung cancer cell line A549 was obtained from PeproTech, Inc. (Rocky Hill, NJ, USA).

Techniques: Transfection, Small Interfering RNA

Journal: Oncology Letters

Article Title: Effects of tanshinones mediated by forkhead box O3a transcription factor on the proliferation and apoptosis of lung cancer cells

doi: 10.3892/ol.2018.9530

Figure Lengend Snippet: Effect of Ts on the proliferation of transfected A549 cells. Cell proliferation was measured using an MTT assay at various time points. A549 cells were transfected with forkhead box O3a siRNA/ssiRNA, treated with various concentrations of Ts, Groups I–IV were treated with 5, 10, 20 and 30 µmol/l respectively. *P<0.05, **P<0.01 vs. control cells treated with ssiRNA. Ts, tanshinones; siRNA, small interfering RNA; ssiRNA, scrambled siRNA; OD, optical density.

Article Snippet: The human lung cancer cell line A549 was obtained from PeproTech, Inc. (Rocky Hill, NJ, USA).

Techniques: Transfection, MTT Assay, Small Interfering RNA

Journal: Frontiers in Immunology

Article Title: A novel oncolytic Vaccinia virus armed with IL-12 augments antitumor immune responses leading to durable regression in murine models of lung cancer

doi: 10.3389/fimmu.2024.1492464

Figure Lengend Snippet: A novel tumor-selective oncolytic VVL-m12 replicates efficiently in and is cytotoxic to lung cancer cell lines. (A) Vaccinia virus Lister strain with deletions of the thymidine kinase (TK) and N1L genes has been described previously . We retained the original B5R gene and placed the signal peptide, stalk (S), transmembrane (T) and cytoplasmic tail (C) regions (STC) in the deleted four short consensus repeat (SCR) domains under an H5 promoter control within the TK region to obtain VV CTRL . IL-12 (murine (m) or human (h)) was incorporated into the N1L region of VV CTRL under control of the H5 promoter, creating VVL-m/h12. (B) Cytotoxicity of VVLΔTKΔN1L, VV CTRL and VVL-m12 against murine lung cancer LLC, CMT64, CMT167 and CMT170 cells and normal murine NIH3T3 cells. Cell death was determined by MTS assay 144 hours post-infection. EC50 values ± SEM are shown. One-way ANOVA with post hoc Tukey tests were used to assess significance (n=3/group). (C, D) Production of infectious virions after infection of murine lung cancer cell lines and normal murine cells over time. Virus production from whole cell lysates was assessed (C) and EEV production was determined via titration of viral supernatant from the same experiments (D) . Mean PFU/cell ± SEM is shown at each time-point and statistical significance determined using two-way ANOVA with Tukey’s multiple comparison post-test (n=3/group). (E) IL-12 expression after infection of murine lung cancer cell lines at an MOI of 1 PFU/cell. Supernatant was collected every 24 hours for 72 hours and assayed for IL-12 by ELISA. Data were normalized to cell number infected and displayed as ng/2×10 5 cells. n=3/group. In all cases, the mean ± SEM is shown. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Article Snippet: Mouse CMT167, as well as CMT170 lung cancer cell line, human lung cancer cell lines H1299 and A549 and non-cancerous cell lines NIH3T3 (murine) and NHBE (human) were obtained from the Centre for Biomarkers & Biotherapeutics, Barts Cancer Institute, Queen Mary University of London, UK.

Techniques: Virus, Control, MTS Assay, Infection, Titration, Comparison, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Bovine lactoferrin drives cell cycle arrest and alters the transcriptomic profile of NSCLC cells

doi: 10.1038/s41598-025-13796-5

Figure Lengend Snippet: Effects of bLF on proliferation, viability, morphology and proliferation of A549 cells in vitro. (A) The CCK-8 assay was performed to determine the viability of A549 and H-1299 cells after treatment with different doses of bLF for 24, and 48 h. (B) Microscopic observation of A549 cell morphology after 12 h and 24 h of treatment with different doses of bLF. (C) Colony formation assay of cell proliferation. The bLF concentrations were 0, 0.5, 1.0 and 1.5 mg/mL. (D) Quantitative results of colony formation. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: A549 cells were maintained in F12 medium (Lanzhou Minhai Bioengineering Co. Lanzhou, China) containing 10% FBS (Lanzhou Minhai Bioengineering Co, Lanzhou, China) and 100 U/mL penicillin-streptomycin (Invitrogen, CA, USA).

Techniques: In Vitro, CCK-8 Assay, Colony Assay

Journal: Scientific Reports

Article Title: Bovine lactoferrin drives cell cycle arrest and alters the transcriptomic profile of NSCLC cells

doi: 10.1038/s41598-025-13796-5

Figure Lengend Snippet: Effects of bLF on cell cycle in A549 cells. (A, B) A549 cell cycle distribution was detected using flow cytometry after treated with different concentrations of bLF. (C) RT-qPCR to detect the expression levels of cycle-related genes (CDK2, P21 and CyclinA1). (D) Western blot to detect the expression levels of cycle-related proteins (CDK2, P21 and CyclinA1). (E) Quantitative analysis of the cell cycle-associated proteins. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: A549 cells were maintained in F12 medium (Lanzhou Minhai Bioengineering Co. Lanzhou, China) containing 10% FBS (Lanzhou Minhai Bioengineering Co, Lanzhou, China) and 100 U/mL penicillin-streptomycin (Invitrogen, CA, USA).

Techniques: Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Bovine lactoferrin drives cell cycle arrest and alters the transcriptomic profile of NSCLC cells

doi: 10.1038/s41598-025-13796-5

Figure Lengend Snippet: bLF inhibits migration of A549 cells. (A) Wound healing assay to detect the effects of bLF on A549 cells. (B) RT-qPCR to detect the expression levels of migration-related genes (E-cadherin, Snail, and MMP-2). (C) Western blot to detect the expression levels of migration-related proteins (E-cadherin, Snail, and MMP-2). (D) Quantitative analysis of migration -associated proteins. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: A549 cells were maintained in F12 medium (Lanzhou Minhai Bioengineering Co. Lanzhou, China) containing 10% FBS (Lanzhou Minhai Bioengineering Co, Lanzhou, China) and 100 U/mL penicillin-streptomycin (Invitrogen, CA, USA).

Techniques: Migration, Wound Healing Assay, Quantitative RT-PCR, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Bovine lactoferrin drives cell cycle arrest and alters the transcriptomic profile of NSCLC cells

doi: 10.1038/s41598-025-13796-5

Figure Lengend Snippet: bLF induces apoptosis in A549 cells and induces A549 apoptosis through the mitochondrial apoptosis pathway. (A) Hoechst 33,342 staining to detect the morphological changes in A549 cells. 1–4: The bLF concentrations were 0, 0.5, 1.0 and 1.5 mg/mL. (B) Flow cytometry assay to determine the apoptosis rate of A549 cells after treated with bLF. (C) The results of mitochondrial membrane potential staining. (D) The results of intracellular ROS levels in A549 cells after bLF treated. 1–4: The bLF concentrations were 0, 0.5, 1.0 and 1.5 mg/mL. (E) RT-qPCR to detect the expression levels of mitochondrial apoptosis-related genes (Caspase-3, P53, Bcl-2 and Bax). (F, H) Western blot to detect the expression levels of mitochondrial apoptosis-related proteins (Caspase-3, Caspase-9, P53, Bcl-2, Bax and cyto-C). (G, I) Quantitative analysis of mitochondrial apoptosis-related proteins. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: A549 cells were maintained in F12 medium (Lanzhou Minhai Bioengineering Co. Lanzhou, China) containing 10% FBS (Lanzhou Minhai Bioengineering Co, Lanzhou, China) and 100 U/mL penicillin-streptomycin (Invitrogen, CA, USA).

Techniques: Staining, Flow Cytometry, Membrane, Quantitative RT-PCR, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Bovine lactoferrin drives cell cycle arrest and alters the transcriptomic profile of NSCLC cells

doi: 10.1038/s41598-025-13796-5

Figure Lengend Snippet: Analysis of mRNA expression difference. (A) Heatmap of Correlation Coefficients for Exp_A549 and Con_A549 Samples. The X and Y axes represent individual samples, with the color indicating the correlation coefficient. Darker colors signify higher correlations, while lighter colors indicate lower correlations. (B) Differential Expression Volcano Plot. The X-axis displays the log2-transformed fold change, and the Y-axis shows the -log10-transformed significance value. Red dots represent up-regulated differentially expressed genes (DEGs), blue dots represent down-regulated DEGs, and gray dots represent non-differentially expressed genes (Non-DEGs). (C) Clustering Heatmap of DEGs. Expression values are depicted in varying colors, with high expression genes colored red and low expression genes colored blue. The intensity of the red color indicates a higher expression level, whereas the intensity of the blue color signifies a lower expression level. (D) RT-qPCR to detect the expression levels of upregulated and downregulated genes. (E) KEGG Pathway Enrichment Analysis of DEGs. The X-axis represents the enrichment ratio, and the Y-axis lists the KEGG Pathways. The bubble size reflects the number of genes annotated to a specific KEGG Pathway, while the color denotes the enrichment significance value. A redder color indicates a smaller significance value. (F) VENN/UpSetR Plot of Gene Expressions. Each circle represents a set of genes, and the overlapping areas of different circles indicate the intersection of these gene sets. The non-overlapping portions signify unique gene sets, and the numbers on the plot represent the gene count in the corresponding regions. (G) Detailed KEGG Pathway Enrichment Analysis of DEGs. Based on kegg_pathway annotation classification, the phyper function in R software was utilized for enrichment analysis to calculate P-values. Subsequently, FDR correction was applied to the P-values to obtain Q-values. Typically, functions with Q-values ≤ 0.05 are considered significantly enriched.

Article Snippet: A549 cells were maintained in F12 medium (Lanzhou Minhai Bioengineering Co. Lanzhou, China) containing 10% FBS (Lanzhou Minhai Bioengineering Co, Lanzhou, China) and 100 U/mL penicillin-streptomycin (Invitrogen, CA, USA).

Techniques: Expressing, Quantitative Proteomics, Transformation Assay, Quantitative RT-PCR, Software