Journal: Frontiers in Oncology

Article Title: Inhibition of SMYD2 Sensitized Cisplatin to Resistant Cells in NSCLC Through Activating p53 Pathway

doi: 10.3389/fonc.2019.00306

Figure Lengend Snippet: The expression level of SMYD2-related proteins and the effects of genetic or chemical manipulation of SMYD2 on the cell growth of CDDP-resistant and parental NSCLC cells. (A) SMYD2, p53, and p53 K370me expression levels were measured in CDDP resistant and parental NSCLC cell lines. β-actin was used as a loading control. (B) Cell viability was measured in BAY-598-treated or SMYD2-knockdown NCI-H460/CDDP cells treated with CDDP at different concentrations for 36 h. Scramble siRNA or DMSO was used as a control. The efficacy of genetic or chemical manipulation of SMYD2 was confirmed by Western blot in NCI-H460/CDDP cells. (C) Cell apoptosis was assessed using Annexin V/PI double staining in BAY-598-treated or SMYD2-knockdown CDDP resistant and parental NCI-H460 cells after treated with CDDP at 10 μM for 48 h. * P < 0.05, compared to corresponding control cells.

Article Snippet: The primary antibodies against SMYD2, p53, Cleaved-PARP, and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA), and the primary antibodies against p53 K370Me was purchased from Immunoway Technology (Plano, TX, USA).

Techniques: Expressing, Control, Knockdown, Western Blot, Double Staining

Journal: Frontiers in Oncology

Article Title: Inhibition of SMYD2 Sensitized Cisplatin to Resistant Cells in NSCLC Through Activating p53 Pathway

doi: 10.3389/fonc.2019.00306

Figure Lengend Snippet: Epigenetic regulation of p53 and its role in CDDP resistance in NSCLC. (A) Cell viability in NCI-H460/CDDP (p53 wide type) and NCI-H1299(p53 deletion) cells, with p53 gene manipulation, which were treated with CDDP at different concentrations for 48 h. Scramble siRNA or mock vector was used as a control. The p53 knock-down or restoration efficacy was confirmed by Western Blot. (B) The p53 reporter activity was measured in CDDP resistant and parental NCI-H460 cells after treated with BAY-598. The relative luciferase unit was calculated by Luciferase/Renilla and DMSO was considered as 100%. (C) The mRNA expression levels of p21, GADD45, and Bax were assessed by real-time RT-PCR in CDDP resistant and parental NCI-H460 cells treated with 10 μM BAY-598. GAPDH was used as a control. (D) Cell apoptosis was assessed using Annexin V/PI double staining in CDDP resistant and parental NCI-H460 cells which were treated with BAY-598 at 10 μM concentrations for 48 h. * P < 0.05, compared to corresponding control cells.

Article Snippet: The primary antibodies against SMYD2, p53, Cleaved-PARP, and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA), and the primary antibodies against p53 K370Me was purchased from Immunoway Technology (Plano, TX, USA).

Techniques: Plasmid Preparation, Control, Knockdown, Western Blot, Activity Assay, Luciferase, Expressing, Quantitative RT-PCR, Double Staining

Journal: Frontiers in Oncology

Article Title: Inhibition of SMYD2 Sensitized Cisplatin to Resistant Cells in NSCLC Through Activating p53 Pathway

doi: 10.3389/fonc.2019.00306

Figure Lengend Snippet: Effects of SMYD2 inhibition and/or CDDP on tumor growth in an CDDP-resistant xenograft model. (A) Tumor volume was measured in NCI-H460/CDDP xenografts treated with AZ505, CDDP, or the combination of AZ505 and CDDP. (B) The p53 and p53 K370me , and cleaved PARP(clv-PARP) expression levels were measured in NCI-H460/CDDP xenograft tumor tissues. β-actin was used as a loading control. * P < 0.05, combined treatment group compared to single treatment group and vehicle control.

Article Snippet: The primary antibodies against SMYD2, p53, Cleaved-PARP, and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA), and the primary antibodies against p53 K370Me was purchased from Immunoway Technology (Plano, TX, USA).

Techniques: Inhibition, Expressing, Control