p53 open reading frame  (Genecopoeia)

Journal: Therapeutic Advances in Medical Oncology

Article Title: The p53/miR-193a/EGFR feedback loop function as a driving force for non-small cell lung carcinoma tumorigenesis

doi: 10.1177/1758835919850665

Figure Lengend Snippet: p53 directly induced miR-193a and epidermal growth factor receptor (EGFR) to form a double-negative feedback loop. (a) Schematic illustrating the putative p53/EGFR-binding sites in the miR-193a promoter. (b) Direct binding of p53 and EGFR to promoter regions of miR-193a as indicated by PCR-based chromatin immunoprecipitation (ChIP) assays. Binding was confirmed by semi-quantitative PCR, followed by gel electrophoresis. (c) Luciferase reporter assays confirmed the suppression of the miR-193a promoter by EGFR, and the promotion of the miR-193a promoter by p53 through the potential binding. (d) qRT-PCR analysis of the miR-193a levels in A549 cells after the knockdown or overexpression of p53 or EGFR. (e) Quantitative RT-PCR showing the miR-193a levels in non-small cell lung carcinoma (NSCLC) cell lines, i.e., A549: p53 WT; H1299: p53 Null. (f, g) Western blot analysis of p53 and EGFR protein levels in A549 and H1299 cells. (f) Representative images, (g) quantitative analysis. (h) Western blot analysis of p53 and EGFR levels in A549 cells transfected with control siRNA, p53 siRNA, p53 siRNA plus miR-193a mimic, control vector, p53 vector, or p53 vector plus miR-193a inhibitor. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: In EGFR and p53 overexpression assays, mammalian expression plasmids encoding EGFR and p53 open reading frame were designed by GeneCopoeia (Germantown, MD, USA).

Techniques: Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Nucleic Acid Electrophoresis, Luciferase, Quantitative RT-PCR, Knockdown, Over Expression, Western Blot, Transfection, Control, Plasmid Preparation

Journal: Therapeutic Advances in Medical Oncology

Article Title: The p53/miR-193a/EGFR feedback loop function as a driving force for non-small cell lung carcinoma tumorigenesis

doi: 10.1177/1758835919850665

Figure Lengend Snippet: Effects of the p53-induced miR-193a and epidermal growth factor receptor (EGFR) double-negative feedback loop on the growth of non-small cell lung carcinoma (NSCLC) xenografts in mice. (a) Representative images of tumors from mice implanted with control A549 cells, miR-193a-overexpressing A549 cells, EGFR-overexpressing A549 cells, miR-193a plus EGFR co-overexpressing A549 cells, p53-inhibiting A549 cells, or miR-193a-overexpressing plus p53-inhibiting A549 cells. A549 cells (1 × 10 6 cells per 0.1 ml) with different treatments were implanted subcutaneously into 6-week-old mice (3 mice per group). (b) The time course of tumor growth in the implanted mice. Tumor volume was measured every 3 days for 24 days after the inoculation. (c) Quantitative RT-PCR analysis of miR-193a levels in tumors from the implanted mice. (d) Western blotting analysis of EGFR and p53 protein levels in tumors from the implanted mice. (e) Representative images and quantitative analysis from hematoxylin and eosin (H&E), Ki67, EGFR and TUNEL staining of tumor sections obtained from the six mouse groups. All data were shown as the mean ± SEM of three separate experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In EGFR and p53 overexpression assays, mammalian expression plasmids encoding EGFR and p53 open reading frame were designed by GeneCopoeia (Germantown, MD, USA).

Techniques: Control, Quantitative RT-PCR, Western Blot, TUNEL Assay, Staining

Journal: Therapeutic Advances in Medical Oncology

Article Title: The p53/miR-193a/EGFR feedback loop function as a driving force for non-small cell lung carcinoma tumorigenesis

doi: 10.1177/1758835919850665

Figure Lengend Snippet: Schematic for the p53/miR-193a/ epidermal growth factor receptor (EGFR) feedback loop in non-small cell lung carcinoma (NSCLC).

Article Snippet: In EGFR and p53 overexpression assays, mammalian expression plasmids encoding EGFR and p53 open reading frame were designed by GeneCopoeia (Germantown, MD, USA).

Techniques: