human embryonic lung fibroblasts cell line  (ATCC)

Journal: Biomolecules

Article Title: Lumbrokinase Extracted from Earthworms Synergizes with Bevacizumab and Chemotherapeutics in Treating Non-Small Cell Lung Cancer by Targeted Inactivation of BPTF/VEGF and NF-κB/COX-2 Signaling

doi: 10.3390/biom14070741

Figure Lengend Snippet: Lumbrokinase induced cellular apoptosis and inhibited proliferation and metastasis in NSCLC cells. ( A ) H460, H1299, A549, and HFL1 cells were treated with different concentrations of lumbrokinase for 48 h, and then the cell viability was tested by MTT assay. IC50 values were calculated by CVXPT32. ( B ) The colony formation assay of H460 and H1299 cells treated with lumbrokinase was detected. ( C ) H460 and H1299 cells were treated with two doses of lumbrokinase for 48 h, and the expressions of the key proteins in the MAPK/Erk signaling pathway were detected by Western blot. ( D ) The expressions of the key proteins in the PI3K/Akt signaling pathway were detected in H460 and H1299 cells with lumbrokinase treatment by Western blot. ( E ) The apoptosis-related proteins were detected by Western blot in H460 and H1299 cells treated with lumbrokinase. ( F ) FC analysis was used to detect cell apoptosis in H460 and H1299 cells treated with lumbrokinase for 48 h. The percentage of apoptotic cells was further calculated. ( G ) Wound-healing assay of H1299 and A549 cells treated with two doses of lumbrokinase for 12 and 24 h was detected (magnification 40×). The cell migration rate was calculated through the quantification of migration distance. ( H ) The invasive capacity of H460 and H1299 cells treated with two doses of lumbrokinase for 48 h was analyzed by transwell assay. The invaded cells were stained with crystal violet and the representative images were taken with an inverted microscope (magnification 40×). ( I ) The EMT-related proteins were detected by Western blot in H460 and H1299 cells treated with lumbrokinase. The data are presented as the mean ± SD. The level of significance was indicated by * p < 0.05, ** p < 0.01, and *** p < 0.001, and ns means no statistical significance.

Article Snippet: Human non-small cell lung cancer cell lines H460, H1299, and A549, human embryonic lung fibroblasts HFL1, and human umbilical vein endothelial cell HUVEC were purchased from ATCC in the United States.

Techniques: MTT Assay, Colony Assay, Western Blot, Wound Healing Assay, Migration, Transwell Assay, Staining, Inverted Microscopy

Journal: Cell Adhesion & Migration

Article Title: Galectin-1 overexpression induces normal fibroblasts translate into cancer-associated fibroblasts and attenuates the sensitivity of anlotinib in lung cancer

doi: 10.1080/19336918.2024.2335881

Figure Lengend Snippet: Galectin-1 overexpression in HFL1 cells. (a) Fluorescence images presented that galectin-1 was successfully overexpression in HFL1 cells (magnification: ×100). (b) qRT-PCR was conducted to measure galectin-1 mRnas expression. (c) Western blotting was conducted to measure galectin-1, TGF-β1 proteins. (d) The qualification of galectin-1, TGF-β1 proteins expression in ctrl and galectin-1-OE groups at different time. (e) Western blotting was conducted to measure α-SMA proteins expression in ctrl and galectin-1-OE groups at different time. The quantitative analysis of galectin-1 (f), TGF-β1 (g), and α-SMA (h) proteins. * p < .05, ** p < .01, *** p < .001.

Article Snippet: Galectin-1 overexpression was conducted in Human embryonic lung fibroblasts (HFL1) cell.

Techniques: Over Expression, Fluorescence, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell Adhesion & Migration

Article Title: Galectin-1 overexpression induces normal fibroblasts translate into cancer-associated fibroblasts and attenuates the sensitivity of anlotinib in lung cancer

doi: 10.1080/19336918.2024.2335881

Figure Lengend Snippet: Galectin-1 overexpression promotes tumor growth in vivo . (a) Imaging technology to monitor the growth of A549 ×enograft tumors. (b)The growth of xenograft tumors was determined by volume. (c) The growth of xenograft tumors was determined by tumor weight. vs A549 group, ** p < .01; vs A549+Anlotinib group, ## p < .01; vs gal-1 OE@HFL1:A549 group, && p < .01.

Article Snippet: Galectin-1 overexpression was conducted in Human embryonic lung fibroblasts (HFL1) cell.

Techniques: Over Expression, In Vivo, Imaging

Journal: Cell Adhesion & Migration

Article Title: Galectin-1 overexpression induces normal fibroblasts translate into cancer-associated fibroblasts and attenuates the sensitivity of anlotinib in lung cancer

doi: 10.1080/19336918.2024.2335881

Figure Lengend Snippet: IHC detected α-SMA, and Ki67 protein expression in vivo . (a) IHC staining of α-SMA protein in xenograft tumors. (b) The staining index of α-SMA. (c) IHC staining of Ki67 protein in xenograft tumors. (d) Positive cell percent of Ki67. Scale bar: 50 μm. (e) Western blotting was conducted to measure MMP2, MMP9, TGF-β1, VEGFA, PDGFA. (f) The qualification of proteins expression in different groups. vs A549 group, * p < .05, ** p < .01; vs A549+Anlotinib group, # p < .05, ## p < .01; vs gal-1 OE@HFL1:A549 group, & p < .05, && p < .01.

Article Snippet: Galectin-1 overexpression was conducted in Human embryonic lung fibroblasts (HFL1) cell.

Techniques: Expressing, In Vivo, Immunohistochemistry, Staining, Western Blot

Journal: Biomedicines

Article Title: Ιnterleukin-17A-Enriched Neutrophil Extracellular Traps Promote Immunofibrotic Aspects of Childhood Asthma Exacerbation

doi: 10.3390/biomedicines11082104

Figure Lengend Snippet: Human embryonic lung fibroblasts (HELFs) acquire a pro-fibrotic phenotype upon treatment with in vitro-generated asthma exacerbation NETs. HELFs were incubated with NETs released from control neutrophils upon stimulation with asthma serum (asthma NETs). ( A ) Migration/wound healing potential (original magnification, 40×) in HELFs stimulated with asthma NETs, assessed via light microscopy. ( B ) Fluorescence microscopy images showing CCN2 staining (blue, DAPI; green, CCN2; original magnification, 100×) and migration/wound healing capacity in HELFs treated with asthma NETs and ( C ) production of collagen ( n = 6 independent experiments). To hinder IL-17A signaling, asthma NETs were pre-incubated with a neutralizing antibody against IL-17A. DNase I was used to dismantle NETs. For ( A , B ), representative examples of 4 independent experiments are shown. For ( C ), data are shown as mean ± SD, Kruskal–Wallis test. All conditions were compared to untreated (statistically significant: p < 0.05).

Article Snippet: Human embryonic lung fibroblasts (HFL-1, Cat#: CCL-153, American Type Culture Collection, Manassas, WV, USA) were cultured at 37 °C with 5% CO 2 in culture medium, consisting of low-glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA), 10% v / v fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA), 100 U/mL Antibiotic-Antimycotic (Biosera, Cholet, France) and 5% v / v MEM non-essential amino acids solution (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: In Vitro, Generated, Incubation, Control, Migration, Light Microscopy, Fluorescence, Microscopy, Staining

Journal: Regenerative Therapy

Article Title: Human bone marrow mesenchymal stem cell-derived exosomes containing microRNA-425 promote migration, invasion and lung metastasis by down-regulating CPEB1

doi: 10.1016/j.reth.2022.03.007

Figure Lengend Snippet: MiR-425 is up-regulated and CPEB1 is down-regulated in lung cancer tissues; miR-425 targets CPEB1. A. RT-qPCR detection of miR-425 expression in lung cancer tissues (n = 93) and adjacent normal tissues (n = 93); B. RT-qPCR detection of CPEB1 mRNA expression in lung cancer tissues (n = 93) and adjacent normal tissues (n = 93); C-D. Protein bands and protein expression of CPEB1 by Western blot analysis (n = 93); E. Pearson correlation analysis of the relation between miR-425 expression and CPEB1 mRNA expression in lung cancer patients; F. miR-425 and CPEB1 mRNA expression in HFL1 and lung cancer cell lines (A549, NCI-H1299 and NCI-H292); G. Targetscan prediction of the targeting relationship between miR-425 and CPEB1; H. Dual luciferase reporter gene assay validation of the targeting relationship between miR-425 and CPEB1. Data in the figure were expressed by mean ± standard deviation; Comparison between two groups was analyzed by t-test.

Article Snippet: Normal human embryonic lung fibroblast HFL1 and A549 lung cancer cells were purchased from Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China) while human lung cancer cells (NCI-H1299, NCI-H292) were from ATCC (VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Luciferase, Reporter Gene Assay, Biomarker Discovery, Standard Deviation, Comparison