mkn45 stomach cancer cells  (ATCC)

Journal: Oncogene

Article Title: NF-κB-induced NOX1 activation promotes gastric tumorigenesis through the expansion of SOX2-positive epithelial cells

doi: 10.1038/s41388-019-0702-0

Figure Lengend Snippet: The induction of NOXO1 expression in gastric cancer cells by TNF-α/NF-κB pathway. a The relative mRNA levels of NOXO1 in the TNF-α-stimulated gastric cancer cells SNU601, SNU719, MKN45, and KATOIII are shown (mean ± s.d.). * p < 0.05; ** p < 0.01. b Immunoblotting analyses for NOXO1 (left) and phosphorylated p65 (right) in the SNU601 cells after TNF-α stimulation at the indicated time. The relative band intensities to 0 h of TNF-α stimulation are indicated at the top of the panels, along with graphs after normalization with GAPDH (left) or p65 (right). c The relative mRNA levels of RELA (left) and NOXO1 (right) normalized with the levels of GAPDH in the TNF-α-stimulated or unstimulated SNU719 cells are shown (mean ± s.d.). * p < 0.05; N.S. not significant. d A schematic view of the NOXO1 promoter regions (from −2000 bp to the transcription start site [TSS]) showing two putative NF-κB binding sites, indicated by boxes. Mut1 and mut2 vectors contain mutations at either of the two NF-κB binding sites. These fragments were used for a luciferase reporter assay. e The relative luciferase activities in SNU601 cells after TNF-α stimulation at 0, 3, and 6 h are shown (mean ± s.d.). *** p < 0.001. f The relative percent inputs of ChIP-based real-time PCR results for NOXO1 promoter in SNU601 cells after TNF-α stimulation at 0 and 3 h are shown (mean ± s.d.). *** p < 0.001; N.S. not significant

Article Snippet: The stomach cancer cells SNU601, SNU719 (Korean Cell Line Bank, Seoul, Korea), MKN45 and Kato-III (Riken Bioresource Center, Tsukuba, Japan) were used for the experiments.

Techniques: Expressing, Western Blot, Binding Assay, Luciferase, Reporter Assay, Real-time Polymerase Chain Reaction

Journal: Oncogene

Article Title: NF-κB-induced NOX1 activation promotes gastric tumorigenesis through the expansion of SOX2-positive epithelial cells

doi: 10.1038/s41388-019-0702-0

Figure Lengend Snippet: The role of NOXO1 in TNF-α-induced ROS production. a Representative photographs of DHE staining (top) and merged images with DAPI staining (bottom) of wild-type mouse stomach (left), K19-C2mE mouse gastritis-associated hyperplasia (center), and Gan mouse gastric tumors (right). Dashed lines indicate the neck region of K19-C2mE hyperplasia. Scale bars, 250 µm. b Representative photographs of DHE staining (top) and merged images with DAPI (bottom) of SNU601 cells at the indicated time after TNF-α stimulation. Scale bars, 100 µm. c The mean ratio of the SNU601 cells positively stained for DHE at the indicated time after TNF-α stimulation (mean ± s.d.). ** p < 0.01; N.S. not significant. d The relative mRNA levels of NOXO1 in the sh-NOXO1 lentivirus-infected cells compared to the control SNU601 cells are shown. ** p < 0.01. e Representative photographs of DHE staining (top) and merged images with DAPI (bottom) of TNF-α-stimulated sh-NOXO1 lentivirus-infected (right) and TNF-α-stimulated control lentivirus-infected SNU601 cells (left). Scale bars, 100 µm. f The mean ratio of the TNF-α-stimulated sh-NOXO1 and control lentivirus-infected cells positively stained for DHE (mean ± s.d.). * p < 0.05

Article Snippet: The stomach cancer cells SNU601, SNU719 (Korean Cell Line Bank, Seoul, Korea), MKN45 and Kato-III (Riken Bioresource Center, Tsukuba, Japan) were used for the experiments.

Techniques: Staining, Infection, Control

Journal: Oncogene

Article Title: NF-κB-induced NOX1 activation promotes gastric tumorigenesis through the expansion of SOX2-positive epithelial cells

doi: 10.1038/s41388-019-0702-0

Figure Lengend Snippet: The suppression of the SOX2 pathway by NOX inhibition in vitro. a The results of RNAseq are shown as an expression plot of each gene in apocynin-treated and control MKN45 cells. The average log2 (rpkm) values of two trials were plotted. b The results of a GO term analysis for “disease and bio function” using the differentially expressed genes in apocynin-treated MKN45 cells. c Depicted results of an ingenuity pathway analysis (IPA) for SOX2 and its targets with an increased or decreased expression in apocynin-treated MKN45 cells are shown. d The relative mRNA levels of SOX2 and SOX2 -target genes (indicated by asterisks in ( c )) in the apocynin-treated and control (CTRL) MKN45 cells are shown (mean ± s.d.). * p < 0.05; ** p < 0.01; N.S. not significant. e The relative mRNA levels of SOX2 and SOX2 -target genes (indicated by asterisk in ( c )) in the SOX2 -targeted siRNA-transfected (siSOX2#1 and #2) and control (si control) MKN45 cells are shown (mean ± s.d.). * p < 0.05; ** p < 0.01; N.S. not significant. f The relative mRNA levels of SOX2 in the GKT136901-treated and control (CTRL) MKN45 cells are shown (mean ± s.d.). * p < 0.05; ** p < 0.01. g The relative mRNA levels of SOX2 in the staurosporin-treated MKN45 and SNU601 cells and no-drug control (CTRL) are shown (mean ± s.d.). ** p < 0.01; *** p < 0.01

Article Snippet: The stomach cancer cells SNU601, SNU719 (Korean Cell Line Bank, Seoul, Korea), MKN45 and Kato-III (Riken Bioresource Center, Tsukuba, Japan) were used for the experiments.

Techniques: Inhibition, In Vitro, Expressing, Control, Transfection