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: Oncology Reports

Article Title: Heparin-binding epidermal growth factor-like growth factor is a potent regulator of invasion activity in oral squamous cell carcinoma

doi: 10.3892/or.2011.1616

Figure Lengend Snippet: Expression levels of EGF family mRNA in HSC3 cells. Amphiregulin (AREG), epigen (EPHN), epiregulin (EREG), HB-EGF (HBEGF), TGF-α (TGF), betacellulin (BTC), EGF and β-actin mRNA expression levels in HSC3 cells were analyzed by RT-qPCR.

Article Snippet: HSC3 cells were transfected with HB-EGF siRNA oligos (sense 5′-GGACCCAUGUCUUCGGAAA-3′ and antisense 5′-UUUCCGAAGACAUGGGUCC-3′) or control scrambled siRNA oligos (sense 5′-AUCCGCGCGAU AGUACGUA-3′ and antisense 5′-UACGUAGUAUCGCGCG GAU-3′) (B-Bridge International, Mountain View, CA) using Lipofectamine™ RNAiMAX (Invitrogen) according to the manufacturer’s instructions, followed by incubation for 48 h at 37°C in a humidified atmosphere of 5% CO 2 in air.

Techniques: Expressing, Quantitative RT-PCR

Journal: Oncology Reports

Article Title: Heparin-binding epidermal growth factor-like growth factor is a potent regulator of invasion activity in oral squamous cell carcinoma

doi: 10.3892/or.2011.1616

Figure Lengend Snippet: Knockdown of heparin-binding epidermal growth factor-like growth factor (HB-EGF) affects invasion activity in HSC3 cells. (A) Expression of HB-EGF mRNA in HSC3 cells treated with siRNAs was determined by real-time RT-PCR. The amount of HB-EGF mRNA was significantly reduced in HB-EGF siRNA-treated HSC3 cells compared with that in control no-siRNA or scrambled-siRNA-treated HSC3 cells. (B) Effect of HB-EGF on HSC3 cell proliferation. The MTT assay showed similar proliferation rates among cells treated with HB-EGF siRNA (closed circle), no-siRNA (triangle), and scrambled-siRNA (open circle). (C) Invasion activity of HB-EGF siRNA-transfected HSC3 cells as determined by the Matrigel invasion assay. The invasion index represents the ratio of cells migrating through a Matrigel-coated membrane/cells migrating through a control non-coated membrane. No-siRNA and scrambled-siRNA transfectants were highly invasive, whereas the invasiveness of HB-EGF siRNA-transfected HSC3 cells was significantly lower.

Article Snippet: HSC3 cells were transfected with HB-EGF siRNA oligos (sense 5′-GGACCCAUGUCUUCGGAAA-3′ and antisense 5′-UUUCCGAAGACAUGGGUCC-3′) or control scrambled siRNA oligos (sense 5′-AUCCGCGCGAU AGUACGUA-3′ and antisense 5′-UACGUAGUAUCGCGCG GAU-3′) (B-Bridge International, Mountain View, CA) using Lipofectamine™ RNAiMAX (Invitrogen) according to the manufacturer’s instructions, followed by incubation for 48 h at 37°C in a humidified atmosphere of 5% CO 2 in air.

Techniques: Binding Assay, Activity Assay, Expressing, Quantitative RT-PCR, MTT Assay, Transfection, Invasion Assay

Journal: Oncology Reports

Article Title: Heparin-binding epidermal growth factor-like growth factor is a potent regulator of invasion activity in oral squamous cell carcinoma

doi: 10.3892/or.2011.1616

Figure Lengend Snippet: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) upregulates MMP-9 expression. (A) MMP-9 mRNA expression and collagenase activity in siRNA-treated HSC3 cells was determined by RT-PCR and zymography, respectively. The MMP-9 mRNA level was reduced in HB-EGF siRNA-treated cells compared with the levels in control no-siRNA and scrambled-siRNA-treated HSC3 cells. (B) The addition of s-HB-EGF to the culture medium upregulated MMP-9 mRNA levels in HSC3 cells.

Article Snippet: HSC3 cells were transfected with HB-EGF siRNA oligos (sense 5′-GGACCCAUGUCUUCGGAAA-3′ and antisense 5′-UUUCCGAAGACAUGGGUCC-3′) or control scrambled siRNA oligos (sense 5′-AUCCGCGCGAU AGUACGUA-3′ and antisense 5′-UACGUAGUAUCGCGCG GAU-3′) (B-Bridge International, Mountain View, CA) using Lipofectamine™ RNAiMAX (Invitrogen) according to the manufacturer’s instructions, followed by incubation for 48 h at 37°C in a humidified atmosphere of 5% CO 2 in air.

Techniques: Binding Assay, Expressing, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Zymography

Journal: Oncology Reports

Article Title: Heparin-binding epidermal growth factor-like growth factor is a potent regulator of invasion activity in oral squamous cell carcinoma

doi: 10.3892/or.2011.1616

Figure Lengend Snippet: Shedding of HB-EGF upregulates MMP-9 expression through EGFR receptor activation. (A) The TACE inhibitor TAPI-2 reduced MMP-9 mRNA levels in HSC3 cells. (B) The EGFR inhibitor AG1478 downregulated MMP-9 mRNA levels in HSC3 cells

Article Snippet: HSC3 cells were transfected with HB-EGF siRNA oligos (sense 5′-GGACCCAUGUCUUCGGAAA-3′ and antisense 5′-UUUCCGAAGACAUGGGUCC-3′) or control scrambled siRNA oligos (sense 5′-AUCCGCGCGAU AGUACGUA-3′ and antisense 5′-UACGUAGUAUCGCGCG GAU-3′) (B-Bridge International, Mountain View, CA) using Lipofectamine™ RNAiMAX (Invitrogen) according to the manufacturer’s instructions, followed by incubation for 48 h at 37°C in a humidified atmosphere of 5% CO 2 in air.

Techniques: Expressing, Activation Assay