human pancreatic adenocarcinoma cells aspc 1  (ATCC)

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 inhibits PAAD proliferation, migration, and invasion. A, qRT-PCR assay results: the LEF1 mRNA level was lower than that in sh-NC after transfection with sh-LEF1. B and C, WB and quantitative analysis: the LEF1 protein level decreased after transfection with sh-LEF1. D and E, CCK-8 assay: the viability decreased after LEF1 knockdown. F and G, EdU incorporation results and quantitative analysis: the number of proliferating cells of AsPC-1/sh-LEF1 and BxPC-3/sh-LEF1 decreased. H and I, Clone formation assay and clone number counting: the number of clones formed by AsPC-1 and BxPC-3 decreased after LEF1 knockdown. J and K, Wound healing assay and mobility calculations: the abilities of migration decreased after LEF1 knockdown. L and M, Transwell assay and counting of the number of cells crossing the basement membrane: the invasion ability of AsPC-1 and BxPC-3 decreased after LEF1 knockdown. N–P, WB and quantitative analysis: the levels of PCNA, MMP2, and MMP9 proteins decreased in AsPC-1 and BxPC-3 after LEF1 knockdown (n = 3, * P <0.05, ** P <0.01).

Article Snippet: Human pancreatic ductal epithelial cells HPDE6-C7, human pancreatic adenocarcinoma cells AsPC-1, BxPC-3, PANC-1, PaTu-8988, and human vascular endothelial cells HUVEC were purchased from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Migration, Quantitative RT-PCR, Transfection, CCK-8 Assay, Tube Formation Assay, Clone Assay, Wound Healing Assay, Transwell Assay, Membrane

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and Cleaved caspase-9 increased (n = 3, * P <0.05, ** P <0.01).

Article Snippet: Human pancreatic ductal epithelial cells HPDE6-C7, human pancreatic adenocarcinoma cells AsPC-1, BxPC-3, PANC-1, PaTu-8988, and human vascular endothelial cells HUVEC were purchased from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Flow Cytometry, Staining, Lactate Dehydrogenase Assay

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 inhibits neovascularization and enhances immune response. A and B, CCK-8 assay showed that conditioned medium from AsPC-1/LEF1 and BxPC-3/LEF1 did not effect HUVEC proliferation. C–E, Transwell assay results and migrating cell number counts showed that AsPC-1/LEF1 and BxPC-3/LEF1 conditioned medium inhibited HUVEC migration. F–H, Tubule formation assay results and tube structure statistics showed that AsPC-1/LEF1 and BxPC-3/LEF1 conditioned medium inhibited HUVEC angiogenic ability and decreased the number of tube structures. I and J, WB assay results and quantitative analysis showed that AsPC-1/LEF1 and BxPC-3/LEF1 conditioned medium downregulated the VEGFA protein level in HUVEC cells. K–N, ELISA results and content statistics showed that knockdown of LEF1 downregulated the contents of TGF-β1, IL-10, PD-L1, and CCL2 in AsPC-1 and BxPC-3 cells (n = 3, * P <0.05).

Article Snippet: Human pancreatic ductal epithelial cells HPDE6-C7, human pancreatic adenocarcinoma cells AsPC-1, BxPC-3, PANC-1, PaTu-8988, and human vascular endothelial cells HUVEC were purchased from the American Type Culture Collection (ATCC).

Techniques: Knockdown, CCK-8 Assay, Transwell Assay, Migration, Tube Formation Assay, Enzyme-linked Immunosorbent Assay

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 interferes with PAAD cell malignant progression and neovascularization by inhibiting Notch1 and NF-κB signaling pathways. A–F, Immunofluorescence experiments and fluorescence intensity analysis: the relative fluorescence level of Notch1 and P65 decreased in AsPC-1/LEF1 and BxPC-3/LEF1. G–M, WB assay results and quantitative analysis: the total Notch1, NICD, Hes1, Hey1, and Jagged1 protein levels and the p-P65/P65 ratio were decreased in AsPC-1/LEF1 and BxPC-3/LEF1 (n = 3, * P <0.05).

Article Snippet: Human pancreatic ductal epithelial cells HPDE6-C7, human pancreatic adenocarcinoma cells AsPC-1, BxPC-3, PANC-1, PaTu-8988, and human vascular endothelial cells HUVEC were purchased from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Protein-Protein interactions, Immunofluorescence, Fluorescence