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:

Journal: Cancer Biology & Therapy

Article Title: Targeting long non-coding RNA PVT1/TGF-β/Smad by p53 prevents glioma progression

doi: 10.1080/15384047.2022.2042160

Figure Lengend Snippet: P53 targets lncRNA PVT1. (a), p53 expression determined by RT-qPCR in clinical samples of glioma patients (N = 75) and normal samples (N = 10). (b), p53 expression determined by RT-qPCR in glioma cell lines. (c), Correlation of p53 expression with patients’ survival analyzed by the Kaplan-Meier method (N = 75). (d), Pearson correlation of p53 expression with lncRNA PVT1 expression in clinical samples of patients with glioma (N = 75). (e), Transfection efficiency of oe-p53 and siRNA-p53 determined by RT-qPCR in glioma cells. (f), LncRNA PVT1 expression determined by RT-qPCR in glioma U373 cells following p53 overexpression. (g), LncRNA PVT1 expression determined by RT-qPCR in glioma U373 cells following p53 knockdown. (h), Interaction between lncRNA PVT1 and p53 confirmed by RIP assay in glioma cells. (i), Binding sites between lncRNA PVT1 promoter and p53 predicted on lncATLAS website. (j), Binding of p53 to lncRNA PVT1 promoter confirmed by dual-luciferase reporter gene assay in 293 T cells. * p < .05. The experiment was repeated 3 times independently.

Article Snippet: The sequence of siRNA targeting p53 (SR322075), consisting of siRNA-control (sense: 5’-ACUACUGAGUGACAGUAGA-3’, antisense: 5’-UCUACUGUCACUCAGUAGU-3’) and siRNA-p53 (sense: 5’-UGCGUGUGGAGUAUUUGGAUG-3’, antisense: 5’-UGGUACAGUCAGAGCCAACCUC-3’), was designed by OriGene Company.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Over Expression, Binding Assay, Luciferase, Reporter Gene Assay

Journal: Cancer Biology & Therapy

Article Title: Targeting long non-coding RNA PVT1/TGF-β/Smad by p53 prevents glioma progression

doi: 10.1080/15384047.2022.2042160

Figure Lengend Snippet: P53 targeting lncRNA PVT1 restrains glioma cell proliferation, migration, and invasion, whereas inducing apoptosis in vitro . (a), Transfection efficiency of siRNA-PVT1 and siRNA-p53 determined by RT-qPCR in U373 cells. (b), U373 cell viability measured by CCK-8 assay. (c), U373 cell proliferation measured by EdU assay. (d), U373 cell cycle distribution detected by flow cytometry. (e), U373 cell migration and invasion measured by Transwell assay. (f), Expression of N-cadherin, MMP3, E-cadherin, and MMP9 proteins in U373 cells determined by Western blot analysis. (g), U373 cell apoptosis measured by flow cytometry. * p < .05. The experiment was repeated 3 times independently.

Article Snippet: The sequence of siRNA targeting p53 (SR322075), consisting of siRNA-control (sense: 5’-ACUACUGAGUGACAGUAGA-3’, antisense: 5’-UCUACUGUCACUCAGUAGU-3’) and siRNA-p53 (sense: 5’-UGCGUGUGGAGUAUUUGGAUG-3’, antisense: 5’-UGGUACAGUCAGAGCCAACCUC-3’), was designed by OriGene Company.

Techniques: Migration, In Vitro, Transfection, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Flow Cytometry, Transwell Assay, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Antipsychotic Drug Aripiprazole Protects Liver Cells from Oxidative Stress

doi: 10.3390/ijms23158292

Figure Lengend Snippet: DNA damage response. Logarithmic fold-change in mRNA expression of long-term treated Fao of ( a ) p53 ( n = 3) ; ( b ) Cdkn1a/p21 ( n = 4), ( c ) Gadd45a ( n = 3), ( d ) Gadd45β ( n = 4), and ( e ) Gadd45 γ ( n = 3). Data are presented as the mean ± standard deviation (SD) and analyzed with one-way ANOVA followed by the Dunnett’s test, comparing all samples to the baseline control with no added H 2 O 2 (*); samples with 1.5 mM H 2 O 2 added are also compared to their untreated control at 1.5 mM H 2 O 2 ( + p ≤ 0.05); samples with 3 mM H 2 O 2 added are compared to their untreated control at 3 mM H 2 O 2 ( # p ≤ 0.05, ## p ≤ 0.01, ### p ≤ 0.001). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; n : number of biological replicates.

Article Snippet: The following TaqMan probes labeled with the FAM dye (Thermo Fisher Scientific) were used: the reference gene Rn18s (Rn03928990_g1), Nrf2/Nfe2l2 (Rn00582415_m1), Srxn1 (Rn04337926_g1), Sirt1 (Rn01428096_m1), Foxo3α (Rn01441087_m1), Bip/Hspa5 (Rn00565250_m1), Chop/Ddit3 (Rn00492098_g1), p53 (Rn00755717_m1), p21 (Rn00589996_m1), Gadd45α (Rn00577049_m1), Gadd45 β (Rn01452530_g1), Gadd45 γ (Rn01352550_g1), Icam1 (Rn00564227_m1), Hspd1 (Rn01441529_g1), Txn2 (Rn00584162_g1), Clpp (Rn01527475_m1), Ymel1l (Rn00586650_m1).

Techniques: Expressing, Standard Deviation, Control

Journal: International Journal of Biological Sciences

Article Title: Bone morphogenetic protein 4 (BMP4) signaling in retinoblastoma cells

doi:

Figure Lengend Snippet: Induction of Id1-3 ( A ), endogenous expression of p21 and p53 ( B ), and expression of p53 ( C ), p21 ( D ), p27 ( E ), p107 ( F ) and p130 ( G ) upon recombinant BMP4 administration to cell cultures 24hrs after administration of 40ng/ml recombinant BMP4. Messenger RNA expression levels of Id1, Id2 and Id3 are significantly up-regulated in WERI-Rb-1 ( A ) upon a BMP4 administration, whereas no changes in p53, p21, p27, p107 and p130 expression level were detectable ( C-G ). Endogeneous levels of p21 and p53 are, however, elevated in WERI-Rb1 cells ( B ). Data were gained from real-time PCR analyses of three independent experiments. Cells that were only treated with 0.1% BSA in 4mM HCl, the solvent for recombinant human BMP4, were taken as a control (ctr.) group for A and C-G , healthy human retina was used as a reference in B and the expression level was set as 1, respectively.

Article Snippet: The following Taqman Gene Expression Assays (Applied Biosystems) were used: Id1: ID Hs00357821_g1; Id2: ID Hs00747379_m1; Id3: ID Hs00171409_m1; p21 (CDKN1A): ID Hs99999142_m1; p27: ID Hs00153277_m1; p53: ID Hs00153340_m1; p130: ID Hs00180562_m1; p107: ID Hs00765707_m1; BMP4: ID Hs00181626_m1.

Techniques: Expressing, Recombinant, RNA Expression, Real-time Polymerase Chain Reaction, Solvent, Control