Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: Effects of GOFA on U937 and HCT116 cells viability (MTT test). ( a ) U937 cells were treated with 1, 10, 25, 50, and 100 μM of GOFA for 24 h. Results are shown as percentages of control samples. Data are presented as means ± SD of 3 independent experiments ( n = 3); ( b ) HCT116 cells were treated with 0.1, 1, 10, 50 and 100 μM of GOFA for 24 h. Data are presented as percentage of control samples (means ± SD, n = 3). * p < 0.05 vs. control cell; # p < 0.05 vs. cells treated with lipopolysaccharide (LPS) alone.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Control

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: Flow cytometry analysis for cell cycle DNA content in G 0 /G 1 , S, G 2 /M phase after treatment of LPS with and w/o GOFA in U937 and HCT116 cells. ( a ) U937 cells were treated with GOFA and the combination of LPS+GOFA and after 24 h cells were stained with propidium iodide for cell cycle analysis and detected by flow cytometry. Percentage of cells was calculated by using MultiCycle software; ( b ) HCT116 cells were treated with GOFA and the combination of LPS+GOFA and after 24 h treatment, cells were processed as described earlier and cell cycle analysis were performed by Flow cytometry. The figures shown are representative analysis from at least three experiments. * p < 0.05 vs. control cells; # p < 0.05 vs. cells treated with LPS alone.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Flow Cytometry, Staining, Cell Cycle Assay, Software, Control

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: Determination of GOFA effect on U937 and HCT116 cell migration. ( a ) Flow cytometric density dot plots (FSC/SSC) of U937 cells, show that cell populations are consistent in the bottom chamber when cultures are stimulated with LPS and treated with GOFA (10 and 50 µM) for 24 h (upper panel). The numerical representation of the cell flow/min in the different experimental conditions is displayed in the lower panel. As shown, the cell count is significantly augmented after the LPS stimulation. In parallel, the number of cells clearly decreases in the presence of loading concentrations of GOFA. The values are presented as the mean ± SD of three independent experiments. * p < 0.05 vs. control cells and # p < 0.05 vs. LPS treated cells; ( b ) representative images of scratch assay for HCT116. Cell monolayer was wounded by a 200 µL pipette tip followed by treatment with various concentrations (0.1, 1 and 10 µM) of GOFA for 24 and 48 h after stimulation with LPS. Scale bar = 100 μm (Upper panel). Scratch closure was evaluated measuring the remaining cell-free area and expressed as percentage of the initial cell-free area (lower panel). # p < 0.05 vs. LPS treated cells at the same time.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Migration, Cell Counting, Control, Wound Healing Assay, Transferring

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: GOFA effects on apoptosis and cell senescence. The effect of GOFA on apoptosis of U937 ( a ) and HCT116 ( b ) cells. The cells were treated with indicated concentration of GOFA for 1 h and then exposed to LPS for 24 h. An Annexin V assay was used for apoptosis detection. ( c ) Bar diagram showing specific effect of GOFA on caspase 3 fluorescent activity on LPS-treated U937 and ( d ) HCT116 cells; ( e ) evaluation of cellular senescence effects of GOFA on U937 and ( f ) HCT116 cells. Cellular senescence was measured using the cellular senescence assay, in which the SA-β-Gal activity was normalized to total protein concentration. All values represent the mean ± SD of three independent experiments. * p < 0.05 vs. control cells; # p < 0.05 vs. cell treated with LPS alone.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Concentration Assay, Annexin V Assay, Activity Assay, Protein Concentration, Control

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: GOFA effects on MMP-9 for U937 ( a ) and HCT-116 ( b ) cell lines. Cell-free conditioned media were assayed for MMP-2 and MMP-9 activity by gelatin zymography. The activity of MMP-9 (92 kDa) (bottom) was reported with fold induction values compared to control MMP-9 activity. Zymogram (top) is representative of 6 gels using 3 separate pools of total protein extracted from cell lines. Changes in MMP-9 gene expression were determined by qRT-PCR assay, using the (ΔΔCt) method and 18S as housekeeping gene. One-way ANOVA, values represent mean ± SD ( n = 6); * p < 0.05 vs. control cells; # p < 0.05 vs. cell treated with LPS alone.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Activity Assay, Zymography, Control, Gene Expression, Quantitative RT-PCR

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: Effects of GOFA on preventing LPS-induced reactive oxygen species (ROS) production in U937 ( a ) and HCT116 ( b ) cells. Antioxidant activity of GOFA against oxidative stress LPS-induced, measured by DCFH-DA assay and compared to ROS scavenger, N-acetyl-L-cysteine (NAC). Results ( n = 3) are reported as means ± SD. * p < 0.05 vs. control cells; # p < 0.05 vs. cell treated with LPS alone.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Antioxidant Activity Assay, DCFH-DA Assay, Control

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: Effects of GOFA on p-p38 and pERK1/2 kinase. Analysis of phosphorylation level of p38 and extracellular signal-regulated kinases (ERK) in GOFA-treated U937 ( a ) and HCT116 ( b ) by Western blotting. Selective inhibitor of p38 (SB203580), ERK (PD980559) and ROS (N-acetyl-L-cysteine, NAC) was used as positive control. Cells were pretreated with selective inhibitors for 30 min and treated with LPS or GOFA + LPS for 24 h. The ratios of p-p38/β-actin and p-ERK/β-actin were calculated based on the relative band densities, respectively. These data are the results of a typical experiment ( n = 3); * p < 0.05 vs. control cells; # p < 0.05 vs. cells treated with LPS alone.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Phospho-proteomics, Western Blot, Positive Control, Control

Journal: Antioxidants

Article Title: Anti-Migratory Effects of 4′-Geranyloxyferulic Acid on LPS-Stimulated U937 and HCT116 Cells via MMP-9 Down-Regulation: Involvement of ROS/ERK Signaling Pathway

doi: 10.3390/antiox9060470

Figure Lengend Snippet: Effects of pharmacological inhibitors on MMP-9 activation and migration in U937 (A) and HCT-116 (B) cell lines. Cells were treated or not with GOFA, selective inhibitor of ERK (PD980559) and ROS (NAC). ( a ) MMP-9 activity by zymography (left) and numerical representation of the cell flow/min (right) in the different experimental conditions of U937 cells. * p < 0.05 vs. control cells; # p < 0.05 vs. LPS treated cells Data are reported as fold induction values (mean ± SD, n = 6). * p < 0.05 vs. cells treated with LPS alone. ( b ) MMP-9 activity by zymography (left) and histogram bar of scratch closure measuring the remaining cell-free area expressed as percentage of the initial cell-free area (right). * p < 0.05 vs. control cells (T 0 ); # p < 0.05 vs. cells treated with LPS alone. The values are presented as the mean ± SD of three independent experiments.

Article Snippet: Human monocytes cell line U937 (ATCC ® CRL-1593.2TM) and human colorectal carcinoma cell line HCT 116 (ATCC ® CCL-247TM) were cultured at a density of 10 6 cells/mL in RPMI 1640 medium (Sigma-Aldrich, MA, USA) and in McCoy’s 5A modified medium (Sigma-Aldrich, MA, USA) respectively.

Techniques: Activation Assay, Migration, Activity Assay, Zymography, Control