Journal: Cancer Management and Research

Article Title: Low-Concentration PTX And RSL3 Inhibits Tumor Cell Growth Synergistically By Inducing Ferroptosis In Mutant p53 Hypopharyngeal Squamous Carcinoma

doi: 10.2147/CMAR.S217944

Figure Lengend Snippet: Ferroptosis contributes to RSL3-induced cell inhibition in mtp53 HPSCC. ( A, B ) Detroit562 and FaDu cells were treated with RSL3 (0.2 µM) for 24 h with or without the pretreatment of Lip-1 (0.02 µM). The cell death was assayed using CCK8 assay. RSL3 was dissolved in DMSO and the cell viability of DMSO was considered as 100%. Data represent the means±SD, n=3 independent experiments. **p<0.01 ***p<0.001 RSL3 vs RSL3+lip-1 treatment group, paired t -test.

Article Snippet: Reagents RSL3 (S7699) and Liproxstatin-1 (S7155) were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Inhibition, CCK-8 Assay

Journal: Cancer Management and Research

Article Title: Low-Concentration PTX And RSL3 Inhibits Tumor Cell Growth Synergistically By Inducing Ferroptosis In Mutant p53 Hypopharyngeal Squamous Carcinoma

doi: 10.2147/CMAR.S217944

Figure Lengend Snippet: Ferroptosis contributes to low-concentration PTX and RSL3-induced synthetic cell death in HPSCC cell lines. ( A ) Quantification of cell viability by CCK8 assay. The effects of low-concentration PTX and RSL3 treatment with or without Lip-1 (Lip-1, 0.02 µM) pretreatment in Detroit562 and FaDu cell lines for 6 and 24 hrs, respectively. ( B ) Representative cell morphological changes are detected by light microscopy (Scale bar=100 µm). Exponentially growing HPSCC cells were transferred to 6-well plates and cultured at 37C in a 5% CO 2 atmosphere. Cells were treated with indicated drugs for 24 hrs. Then, images were taken using an OLYMPUS IX 71 microscope (10×10) (OLYMPUS, Tokyo, Japan). (Data represent the means±SD, n=3 independent experiments. *p<0.05, **p<0.01, paired t -test).

Article Snippet: Reagents RSL3 (S7699) and Liproxstatin-1 (S7155) were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Concentration Assay, CCK-8 Assay, Light Microscopy, Cell Culture, Microscopy

Journal: Cancer Management and Research

Article Title: Low-Concentration PTX And RSL3 Inhibits Tumor Cell Growth Synergistically By Inducing Ferroptosis In Mutant p53 Hypopharyngeal Squamous Carcinoma

doi: 10.2147/CMAR.S217944

Figure Lengend Snippet: Effect of GPX4 overexpression on low-concentration PTX and RSL3-induced ferroptosis. ( A ) Overexpression of GPX4 as demonstrated by Western blot, respectively, in Detroit562 and FaDu cells. Cells were transfected with GPX4 -expressing plasmid (Cat: 718676-1) or its relative negative control for 36 hrs. Protein bands were quantified with Image J. The quantification reflected the relative amounts as a ratio of each protein band relative to the lane’s loading control (GAPDH). ( B ) Detroit562 and FaDu cells were transfected with GPX4 or its negative control was treated with RSL3 (2 µM) and low-concentration PTX (2 nM) for 24 hrs with or without Lip-1 (0.02 µM) pretreatment. Lipid ROS level (Percentage of max) was determined using the C11-BODIPY, samples were examined using BD FACS Calibur. ( C ) Graphical representation of the histogram for relative lipid ROS . ( D ) Detroit562 and FaDu cells were transfected with GPX4 or its negative control was treated with combination of RSL3(2 µM) and low-concentration PTX (2nM) for 24 h. Cell viability was assayed using Cell Counting Kit-8. ( E, F ) Western blotting analysis of GPX4 in Detroit562 and FaDu cells treated with low-concentration PTX (2nM), RSL3 (2 µM), combination drug with or without the pretreatment of Lip-1 (0.02 µM) for 24 hrs. Data represent the means±SD, n=3 independent experiments.**p<0.01, ****p<0.0001, paired t -test

Article Snippet: Reagents RSL3 (S7699) and Liproxstatin-1 (S7155) were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Over Expression, Concentration Assay, Western Blot, Transfection, Expressing, Plasmid Preparation, Negative Control, Control, Cell Counting

Journal: Cancer Management and Research

Article Title: Low-Concentration PTX And RSL3 Inhibits Tumor Cell Growth Synergistically By Inducing Ferroptosis In Mutant p53 Hypopharyngeal Squamous Carcinoma

doi: 10.2147/CMAR.S217944

Figure Lengend Snippet: mtp53 mediated ferroptosis contributes to low-concentration PTX and RSL3-induced synthetic cell death. ( A ) Western blotting analysis of p53 in Detroit562 treated with PTX at indicated low concentrations for 24 hrs. Protein bands were quantified with Image J. The quantification reflected the relative amounts as a ratio of each protein band relative to the lane’s loading control (GAPDH). ( B ) Detroit562 cells were transfected with pCDNA3-GFP-P53 (R175H) plasmid then fixed and stained with anti-P53(R175H) monoclonal antibody (Cat. # 26072) and with secondary antibody. Fluorescent images were collected simultaneously (Scale bar=25 µm). ( C ) Detroit562 and FaDu cells were transfected with p53 siRNA (sc-29435) or its relative negative control was treated with combination of RSL3(2μM) and low-concentration PTX (2 nM) for 24 hrs. Cell viability was assayed using Cell Counting Kit-8. ( D ) Detroit562 and FaDu cells were transfected with p53 siRNA (sc-29435) or its relative negative control was treated with RSL3 (2 µM) and low-concentration PTX (2nM) for 24 hrs with or without Lip-1 (0.02 µM) pretreatment. Lipid ROS level (Percentage of max) was determined using the C11-BODIPY, samples were examined using BD FACS Calibur. ( E ) Graphical representation of the histogram for relative lipid ROS. ( F ) Western blotting analysis of p53 and SLC7A11 in Detroit562 and FaDu cells transfected with p53 siRNA (sc-29435) or its relative negative control. ( G, H ) Western blotting analysis of SLC7A11 in Detroit562 and FaDu cells transfected with p53 siRNA (sc-29435) or its relative negative control. Detroit562 and FaDu cells were treated with low-concentration PTX (2 nM), RSL3 (2 µM), combination drug with or without the pretreatment of Lip-1 (0.02 µM) for 24 hrs. Data represent the means±SD, n=3 independent experiments. ns p>0.05, **p<0.01, ***p<0.001, ****p<0.0001, paired t -test.

Article Snippet: Reagents RSL3 (S7699) and Liproxstatin-1 (S7155) were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Concentration Assay, Western Blot, Control, Transfection, Plasmid Preparation, Staining, Negative Control, Cell Counting