Journal: Antioxidants

Article Title: Thymus zygis subsp. zygis an Endemic Portuguese Plant: Phytochemical Profiling, Antioxidant, Anti-Proliferative and Anti-Inflammatory Activities

doi: 10.3390/antiox9060482

Figure Lengend Snippet: Anti-proliferative effect of Thymus zygis subsp. zygis aqueous decoction (AD) and hydroethanolic (HE) extracts on Caco-2 ( A and B for AD and HE extracts, respectively) and HepG2 cells ( C and D for AD and HE extracts, respectively). Two exposure times, 24 and 48 h, were considered, as indicated. Results are expressed as (mean ± SD, n = 4). Statistically significant differences ( p < 0.05) between the control and sample concentrations at respective incubation time are denoted by *, and those between exposure periods at the same concentration are denoted by #.

Article Snippet: In this study, two human cell lines: Caco-2 (human colon adenocarcinoma cell line; Cell Lines Service, Eppelheim, Germany) and HepG2 (human hepatocellular carcinoma cell line; ATCC ® Number: HB-8065TM, a gift from Prof. C. Palmeira CNC-UC, Portugal) and a mouse cell line: RAW 264.7 (mouse macrophages, Abelson murine leukemia virus-induced tumor cell line; Cell Lines Service, Eppelheim, Germany) cells were used to evaluate the anti-proliferative and anti-inflammatory activities of T. zygis extracts.

Techniques: Control, Incubation, Concentration Assay

Journal: Antioxidants

Article Title: Thymus zygis subsp. zygis an Endemic Portuguese Plant: Phytochemical Profiling, Antioxidant, Anti-Proliferative and Anti-Inflammatory Activities

doi: 10.3390/antiox9060482

Figure Lengend Snippet: Effect Thymus zygis subsp. zygis extracts on Caco-2 and HepG2 cells, expressed as IC 50 values. Cells were exposed to aqueous decoction (AD) and hydroethanolic (HE).

Article Snippet: In this study, two human cell lines: Caco-2 (human colon adenocarcinoma cell line; Cell Lines Service, Eppelheim, Germany) and HepG2 (human hepatocellular carcinoma cell line; ATCC ® Number: HB-8065TM, a gift from Prof. C. Palmeira CNC-UC, Portugal) and a mouse cell line: RAW 264.7 (mouse macrophages, Abelson murine leukemia virus-induced tumor cell line; Cell Lines Service, Eppelheim, Germany) cells were used to evaluate the anti-proliferative and anti-inflammatory activities of T. zygis extracts.

Techniques: Extraction

Journal: Cells

Article Title: miR-200c-3p, miR-222-5p, and miR-512-3p Constitute a Biomarker Signature of Sorafenib Effectiveness in Advanced Hepatocellular Carcinoma

doi: 10.3390/cells11172673

Figure Lengend Snippet: miRNA pattern induced in primary human hepatocytes. ( A ) Heatmap of miRNA expression in HepG2 cell lysates at 6 and 24 h of treatment, compared to primary hepatocyte expression profile after Sorafenib (24 h) (n = 3). ( B ) Volcano plot of miRNA expression analysis in the HepG2 cells compared to primary human hepatocytes (non-treated) (n = 3). Up-regulated miRNAs are shown in red and down-regulated miRNAs are shown in green. ( C ) Volcano plot of differentially expressed miRNAs in primary human hepatocytes treated with Sorafenib (24 h) (n = 3). ( D ) Top 10 most significant KEGG pathways enriched in targets of miRNAs differentially expressed in HepG2 cells. ( E ) Top 10 most significant KEGG pathways enriched in targets of miRNAs differentially expressed in treated primary human hepatocytes.

Article Snippet: The hepatoblastoma HepG2 cell line (HB-8065TM, ATCC-LGC Standards, S.L.U., Barcelona, Spain) [ ] was cultured in MEM with Earle’s salts with L-glutamine with 10% fetal bovine serum (FBS) (Ref. F7524, Sigma-Aldrich, batch BCBX9154, San Luis, CA, USA), sodium pyruvate (1 mM) (Ref. 11360070, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), non-essential amino acids (Ref. 11140035, Gibco, Thermo Fisher Scientific), penicillin–streptomycin solution (100 U/mL-100 μg/mL) (Ref. 15640055, Gibco, Thermo Fisher Scientific) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Expressing

Journal: Cells

Article Title: miR-200c-3p, miR-222-5p, and miR-512-3p Constitute a Biomarker Signature of Sorafenib Effectiveness in Advanced Hepatocellular Carcinoma

doi: 10.3390/cells11172673

Figure Lengend Snippet: Functional analysis with mimics (left panel) and inhibitors (right panel) of differentially expressed miRNAs in HepG2 cells treated with Sorafenib. Proliferation (mimics A , inhibitors B ), caspase-3/7 activity (mimics C , inhibitors D ), migration (mimics E , inhibitors F ), and invasion (mimics G , inhibitors H ) were tested to unravel anti- or pro-tumoral effects of miRNAs after Sorafenib treatment. Invasion was tested for the miRNA mimics and inhibitors that showed the upregulation of cell migration with p -value < 0.01, or those that downregulated cell migration more that 5%. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001 between the transfection control and mimic or inhibitor tested.

Article Snippet: The hepatoblastoma HepG2 cell line (HB-8065TM, ATCC-LGC Standards, S.L.U., Barcelona, Spain) [ ] was cultured in MEM with Earle’s salts with L-glutamine with 10% fetal bovine serum (FBS) (Ref. F7524, Sigma-Aldrich, batch BCBX9154, San Luis, CA, USA), sodium pyruvate (1 mM) (Ref. 11360070, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), non-essential amino acids (Ref. 11140035, Gibco, Thermo Fisher Scientific), penicillin–streptomycin solution (100 U/mL-100 μg/mL) (Ref. 15640055, Gibco, Thermo Fisher Scientific) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Functional Assay, Activity Assay, Migration, Transfection, Control

Journal: Cells

Article Title: miR-200c-3p, miR-222-5p, and miR-512-3p Constitute a Biomarker Signature of Sorafenib Effectiveness in Advanced Hepatocellular Carcinoma

doi: 10.3390/cells11172673

Figure Lengend Snippet: RNA-sequencing data of miR-200c-3p down-regulation in HepG2 cells showed increased damage response mechanisms. ( A ) Venn diagrams showing the mRNAs regulated upon miR-200c-3p inhibition (I-miR-200c-3p) compared to the transfection control (TC). ( B ) Heatmap of genes regulated by miR-200c-3p. ( C ) Ffold-enrichment values of the top five most significantly enriched GO BP, MF, CC, and KEGG pathways. ( D ) Top 10 significant hallmarks GSEA up-regulated after I-miR-200c-3p. Plots were provided for the three most enriched terms. ( E ) Diagram showing the clustering of STRING analysis. ( F ) Fold-enrichment values of the top two most significantly enriched terms in either the GO BP, MF, CC, or KEGG pathways in each STRING cluster. Experiments were carried out as three independent replicates.

Article Snippet: The hepatoblastoma HepG2 cell line (HB-8065TM, ATCC-LGC Standards, S.L.U., Barcelona, Spain) [ ] was cultured in MEM with Earle’s salts with L-glutamine with 10% fetal bovine serum (FBS) (Ref. F7524, Sigma-Aldrich, batch BCBX9154, San Luis, CA, USA), sodium pyruvate (1 mM) (Ref. 11360070, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), non-essential amino acids (Ref. 11140035, Gibco, Thermo Fisher Scientific), penicillin–streptomycin solution (100 U/mL-100 μg/mL) (Ref. 15640055, Gibco, Thermo Fisher Scientific) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: RNA Sequencing, Inhibition, Transfection, Control

Journal: Cells

Article Title: miR-200c-3p, miR-222-5p, and miR-512-3p Constitute a Biomarker Signature of Sorafenib Effectiveness in Advanced Hepatocellular Carcinoma

doi: 10.3390/cells11172673

Figure Lengend Snippet: RNA-sequencing data of miR-222-5p overexpression in HepG2 cells showed altered cell cycle and metabolic control. ( A ) Venn diagrams showing mRNAs regulated upon miR-222-5p mimics (M-miR-222-5p) compared to the transfection control (TC). ( B ) Heatmap of genes regulated by miR-222-5p. ( C ) Fold-enrichment values of the top five most significantly enriched GO BP, MF, CC, and KEGG pathways. ( D ) Top 10 significant hallmarks of GSEA downregulated after M-miR-222-5p. Plots were provided for the three most enriched terms. ( E ) Diagram showing clustering of the STRING analysis. ( F ) Fold-enrichment values of the top two most significantly enriched terms either in the GO BP, MF, CC, or KEGG pathways in each STRING cluster. Experiments were carried out as three independent replicates.

Article Snippet: The hepatoblastoma HepG2 cell line (HB-8065TM, ATCC-LGC Standards, S.L.U., Barcelona, Spain) [ ] was cultured in MEM with Earle’s salts with L-glutamine with 10% fetal bovine serum (FBS) (Ref. F7524, Sigma-Aldrich, batch BCBX9154, San Luis, CA, USA), sodium pyruvate (1 mM) (Ref. 11360070, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), non-essential amino acids (Ref. 11140035, Gibco, Thermo Fisher Scientific), penicillin–streptomycin solution (100 U/mL-100 μg/mL) (Ref. 15640055, Gibco, Thermo Fisher Scientific) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: RNA Sequencing, Over Expression, Control, Transfection

Journal: Cells

Article Title: miR-200c-3p, miR-222-5p, and miR-512-3p Constitute a Biomarker Signature of Sorafenib Effectiveness in Advanced Hepatocellular Carcinoma

doi: 10.3390/cells11172673

Figure Lengend Snippet: RNA-sequencing data of miR-512-3p overexpression in the HepG2 cells showed reduced oxidative metabolism. ( A ) Venn diagrams showing mRNAs regulated upon miR-512-3p mimics (M-miR-512-3p) compared to the transfection control (TC). ( B ) Heatmap of genes regulated by miR-512-3p. ( C ) Fold-enrichment values of the top five most significantly enriched GO BP, MF, CC, and KEGG pathways. ( D ) Top 10 significant hallmarks of GSEA downregulated after M-miR-222-5p. Plots were provided for the three most enriched terms. ( E ) Diagram showing the clustering of STRING analysis. ( F ) Fold-enrichment values of the top two most significantly enriched terms either in the GO BP, MF, CC, or KEGG pathways in each STRING cluster. Experiments were carried out as three independent replicates.

Article Snippet: The hepatoblastoma HepG2 cell line (HB-8065TM, ATCC-LGC Standards, S.L.U., Barcelona, Spain) [ ] was cultured in MEM with Earle’s salts with L-glutamine with 10% fetal bovine serum (FBS) (Ref. F7524, Sigma-Aldrich, batch BCBX9154, San Luis, CA, USA), sodium pyruvate (1 mM) (Ref. 11360070, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), non-essential amino acids (Ref. 11140035, Gibco, Thermo Fisher Scientific), penicillin–streptomycin solution (100 U/mL-100 μg/mL) (Ref. 15640055, Gibco, Thermo Fisher Scientific) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: RNA Sequencing, Over Expression, Transfection, Control