Journal: Redox Biology

Article Title: Induction of 2-hydroxycatecholestrogens O-methylation: A missing puzzle piece in diagnostics and treatment of lung cancer

doi: 10.1016/j.redox.2022.102395

Figure Lengend Snippet: (A) 2-ME significantly decreases A549 cell viability in a dose-dependent manner in the 2D culture model at concentrations less than or equal 100 μM. A549 cells were treated with serial dilutions of 2-ME at a concentration range from 100 μM to 0.43 μM for 24 h. Cell viability was determined by MTT assay. Presented values are the mean ± SE of three independent experiments. *p < 0.01, **p < 0.001, ***p < 0.0001, ****p < 0.00001 vs. control cells. (B) 2-ME reduces the cell viability of the A549 3D spheroidal models in a dose dependent manner with expenditures close to 200 μM. A549 cells were exposed to serial dilutions of 2-ME over a concentration range of 200 μM–0.78 μM for 48 h. Cell viability was determined by the WST-1 assay. Presented values are the mean ± SE of three independent experiments. *p < 0.01, **p < 0.001, ***p < 0.0001, ****p < 0.00001 vs. control cells. (C) IC50 values for the A549 cell line treated with 2-ME were >100 μM. These values were calculated by analyzing the relationship between concentrations and percentage (%) of inhibition using GraphPad Prism version 9.0 for Windows, GraphPad Software, CA, USA.

Article Snippet: Wild-type human lung epithelial carcinoma, A549 cell line as 3D cell culture spheroids were obtained by plating cells with an average density of 8000 cells/cm 2 in 96-well U-bottom and V-bottom Lipidure® cell culture plates (Amsbio, Abingdon, United Kingdom) coated with phosphorylcholine which is naturally found in cell membranes.

Techniques: Concentration Assay, MTT Assay, WST-1 Assay, Inhibition, Software

Journal: Redox Biology

Article Title: Induction of 2-hydroxycatecholestrogens O-methylation: A missing puzzle piece in diagnostics and treatment of lung cancer

doi: 10.1016/j.redox.2022.102395

Figure Lengend Snippet: (A) A549 cells as well as cell medium alone were treated with 2-ME at 10 μM, 1 μM and 10 nM concentrations, respectively. 2-ME treated A549 cells generated higher total hydrogen peroxide levels as compared to untreated control A549 cells incubated with Nutrient Mixture F-12 Ham medium alone. The obtained result comparing ROS induction in the wells with no cells and in the wells with A549 cells, indicates an increased production of ROS by the cancer cells. Luminescence was determined with a Glo-Max® Luminometer from Promega (Mannheim, Germany). Statistical analysis was performed using the GraphPad TTEST function (GraphPad Prism 9 version 9.0.0.). For analysis, both the mean of average luminescence (RLU) and the standard deviation were calculated. (B) Time-dependent growth rate of 2-ME-treated A549 cells. A549 cells treated with 2-ME 10 μM concentration up to 24 h. The analysis of the obtained results evaluated by staining with propidium iodide (PI) was performed using the CytoSMART Analysis System (Lux 3, CytoSMART, Eindhoven, The Netherlands). Values are presented as the mean ± SE of three independent experiments. *p < 0.01, **p < 0.001, ***p < 0.0001, ****p < 0.00001 vs. control cells.

Article Snippet: Wild-type human lung epithelial carcinoma, A549 cell line as 3D cell culture spheroids were obtained by plating cells with an average density of 8000 cells/cm 2 in 96-well U-bottom and V-bottom Lipidure® cell culture plates (Amsbio, Abingdon, United Kingdom) coated with phosphorylcholine which is naturally found in cell membranes.

Techniques: Generated, Incubation, Standard Deviation, Concentration Assay, Staining

Journal: Redox Biology

Article Title: Induction of 2-hydroxycatecholestrogens O-methylation: A missing puzzle piece in diagnostics and treatment of lung cancer

doi: 10.1016/j.redox.2022.102395

Figure Lengend Snippet: (A). A549 cells were pretreated with V5 peptide at a concentration of 100 μM for 4 h, and then treated with 1 μM 2-ME for 24 h. Values are the mean ± SD of three independent experiments. *p < 0.01, **p < 0.001, ***p < 0.0001, ****p < 0.00001 vs. control cells. (B) The cells were treated for 24 h with either 100 μM PalmB or 10 μM 2-ME, and a combination of both. Both PalmB and 2-ME alone, significantly decreased A549 cell viability. A synergistic effect on the viability decrease was observed for treatment with both PalmB and 2-ME used in combination. The cell viability was determined by MTT assay. Values are presented as the mean ± SE of three independent experiments. *p < 0.01, **p < 0.001, ***p < 0.0001, ****p < 0.00001 vs. control cells C. Serum levels of critical estrogen metabolites analyzed in lung cancer patients (LC) as compared with healthy control (C) are presented as median, as well as quartile 1 and 3, with the minimum and maximum values, in the form of a typical box and whiskers plot. Statistically significant differences are marked with an asterisk * and refer to p-value < 0.05. It is worth noting that 2-ME serum level is significantly lower in lung cancer patients as compared with the control group. Based on table No 2.

Article Snippet: Wild-type human lung epithelial carcinoma, A549 cell line as 3D cell culture spheroids were obtained by plating cells with an average density of 8000 cells/cm 2 in 96-well U-bottom and V-bottom Lipidure® cell culture plates (Amsbio, Abingdon, United Kingdom) coated with phosphorylcholine which is naturally found in cell membranes.

Techniques: Concentration Assay, MTT Assay

Journal: Oncotarget

Article Title: High-throughput RNAi screening for novel modulators of vimentin expression identifies MTHFD2 as a regulator of breast cancer cell migration and invasion

doi:

Figure Lengend Snippet: A: The effect of MTHFD2 siRNAs on MDA-MB-231(SA) and BT-549 cell motility in wound healing experiment. Cells were automatically imaged once every hour after wound scratching. Wound closure effect was calculated as wound confluence which the cells gained in twelve hours. B: MDA-MB-231(SA) cells transfected with either vimentin targeting siRNA (siVIM), MTHFD2 targeting siRNA (siMTHFD2) or negative control siRNA (siScr), allowed to invade into Matrigel matrix for 4 days and imaged. Shown are the quantifications of two replicate experiments (imaged at 5-10 different positions; ***p < 0.005) and representative images. The dashed lines indicate the top of the Matrigel matrix.

Article Snippet: BT-549 cells were obtained from Cell Lines Service (Eppelheim, Germany).

Techniques: Transfection, Negative Control