Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: Nthy-ori 3-1 ( a ) and TPC-1 ( b ) cell line viability under H 2 O 2 stress condition. Cell viability was assayed using MTS. The cells were exposed to H 2 O 2 for different time and concentrations (from 50 µM to 10 mM). For each experiment, n = 5 replicates wells were assayed per clone. Cell viability values were calculated as means and compared with untreated quiescent cells (Q). ***— p < 0.001 vs. quiescent; #— p < 0.05; ##— p < 0.01 cells with same dosage of H 2 O 2 treatment at different time.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques:

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: Effect of H 2 O 2 [10 mM] on the cell cycle of starved thyroid cell lines. ( a ) FACS analysis profile of Nthy-ori 3-1 and TPC-1 untreated and treated cells by 10 mM H 2 O 2 after 15 or 30 min; ( b ) relative percentages in cell cycle stages.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques:

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: OGG1 (8-Oxoguanine glycosylase), MUTYH (mutY DNA glycosylase), APE1 (apurinic/apyrimidinic endodeoxyribonuclease-1) and PPARγ (peroxisome-proliferator-activated receptor gamma) gene expression modulation under H 2 O 2 [10 mM], EGF, LY and PD treatments alone and combined in Nthy-ori 3-1 cells ( a ) vs. TPC-1 cells ( b ). Gene expression was analyzed by real-time qPCR. The histogram represented normalized data with GUSB gene in Nthy-ori 3-1, while the PPARγ expression data were normalized with β-Actin in TPC-1 cells. The expression of MUTYH was undetected in TPC-1 cells (five independent experiments). The results showed the average of three independent experiments. LY—LY294002; PD—PD98059; *— p < 0.05; **— p < 0.01; ***— p < 0.001 treated vs. quiescent cells; #— p < 0.05; ##— p < 0.01; ###— p < 0.001 cells with similar treatment.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: EGFR (epidermal growth factor receptor, ERBB2 (Erb-B2 receptor tyrosine kinase 2), ERBB3 (Erb-B3 receptor tyrosine kinase 3) gene expression modulation under H 2 O 2 , EGF, LY and PD treatments alone and combined in Nthy-ori 3-1 cells ( a ) vs. TPC-1 cells ( b ). Gene expression was analyzed by Real-time qPCR. The histogram represented normalized data with GUSB gene, and the results showed the average of three independent experiments. LY—LY294002; PD—PD98059; *— p < 0.05, **— p < 0.01 ***— p < 0.001 treated vs. quiescent cells; #— p < 0.05, ###— p < 0.001 cells with similar treatment.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: NRF2 (nuclear factor erythroid 2–related factor 2), HO1 (heme oxygenase 1), JUN/AP1 (Jun proto-oncogene, AP-1 transcription factor subunit) gene expression and IL8 ( CXCL8 , interleukin 8) modulation under H 2 O 2 , EGF, LY and PD treatments alone and combined in Nthy-ori 3-1 cells ( a ) vs. TPC-1 cells ( b ). Gene expression was analyzed by real-time qPCR. The histogram represented normalized data with GUSB gene in Nthy-ori 3-1, while NRF2 and HO1 expression data were normalized with β-Actin in TPC-1 cells. The results showed the average of three independent experiments. LY—LY294002; PD—PD98059; *— p < 0.05; **— p < 0.01; ***— p < 0.001 treated vs. quiescent cells; #— p < 0.05, ##— p < 0.01, ###— p < 0.001 cells with similar treatment.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: Gene expression modulation of TPO (thyroid peroxidase) and ZEB-1 (zinc finger E-box binding homeobox 1) under H 2 O 2 , EGF, LY and PD treatments alone and combined. ( a ) Nthy-ori 3-1 cells, ( b ) TPC-1 cells. Gene expression was analyzed by real-time qPCR. The histogram represented normalized data with GUSB gene, and the results showed the average of three independent experiments. The expression of TPO was undetected in TPC-1 cells. LY—LY294002; PD—PD98059; *— p < 0.05 treated vs. quiescent cells; #— p < 0.05 cells with similar treatment.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing, Binding Assay

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: MUTYH and OGG1 gene dosage. ( a ) Nthy-ori 3-1 cells, ( b ) TPC-1 cells. Gene dosage was performed by SYBR Green qRT-PCR. For both genes were selected two different genomic sequences. The samples were amplified in triplicate in three independent experiments. The gDNA amounts of the target genes were normalized by the ratio on median value of the b-Actin genomic reference gene from TPC-1 vs. Nthy-ori 3-1.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: SYBR Green Assay, Quantitative RT-PCR, Genomic Sequencing, Amplification

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: Gene expression cluster analysis by MeV4.9.0 in Nthy-ori 3-1 cells: ( a ) H 2 O 2 treatments; ( b ) EGF treatments; ( c ) LY treatments; ( d ) PD treatments. The average linkage hierarchical clustering with Pearson correlation was used. The color scale at the top represents the log2 of every single gene expression value compared with housekeeping value ranging from −13 (green) to 4 (red). The trees presented here are the neighbor-joining trees based on gene expression variation in response to different treatments. LY—LY294002; PD—PD98059.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: Protein expression and quantification in Nthy-ori 3-1 vs. TPC-1 cells. Nthy-ori 3-1 and TPC-1 cells were starved overnight and treated with, LY294002 (25 μM), PD98059 (50μM), EGF (50 ng/mL) alone and combined with H 2 O 2 (10 mM). Western blotting analysis in Nthy-ori 3-1 cells ( a ) and in TPC-1 cells ( b ) determining the protein expression levels of p-EGFR, ErbB2, p-MAPK, p-AKT and OGG1. Results are representative of three independent experiments. Quiescent cells were treated with H 2 O 2 and LY, PD, EGF alone or combined. LY—LY294002; PD—PD98059; kDa—protein molecular weight marker.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing, Western Blot, Molecular Weight, Marker

Journal: Cells

Article Title: Emerging Role of Oxidative Stress on EGFR and OGG1-BER Cross-Regulation: Implications in Thyroid Physiopathology

doi: 10.3390/cells11050822

Figure Lengend Snippet: Protein expression quantification of ErbB pathway NThy-ori 3-1 vs. TPC-1 cells. Quantification of selected proteins (p-EGFR, ErbB2, p-MAPK, p-AKT) detected by WB in Nthy-ori3-1 and TPC-1 ( a,b). The expression of p-EGFR, was undetected in TPC-1 cells. The average expression levels of panel ( a ) and panel ( b ) were determined by densitometric analysis and calculated in relation to the β-Actin level. Quiescent cells were treated with H 2 O 2 and LY, PD, EGF alone or combined. LY—LY294002; PD—PD98059.

Article Snippet: Human thyroid follicular epithelial Nthy-ori 3-1 cells, obtained from European Collection of Authenticated Cell Cultures (ECACC 90011609) (Public Health England, Porton Down, Salisbury, UK) (Sigma Aldrich, St. Louis, MO, USA) were cultured at 37 °C in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin/streptomycin and 2 mM L-glutamine.

Techniques: Expressing

Journal: Life Science Alliance

Article Title: tRF-1:30-Gly-CCC-3 inhibits thyroid cancer via binding to PC and modulating metabolic reprogramming

doi: 10.26508/lsa.202302285

Figure Lengend Snippet: (A) The workflow of tRF and tiRNA sequencing and analysis. (B) The principal component analysis of tRF and tiRNA expression in four pairs of PTC tissues (T) and adjacent normal tissues (AT). (C) The correlation coefficient analysis of tRF and tiRNA expression in tumor tissues and ATs. (D) Venn diagram of tRFs and tiRNAs detected only in tumor tissues (red region), only in ATs (blue region), and in both tissues (purple region). (E) Distributions of different tRFs and tiRNAs in tumor tissues and ATs. (F) Heatmap of differentially expressed tRFs and tiRNAs in the two groups. (G) Volcano plot of down-regulated tRFs and tiRNAs (green dots) and up-regulated tRFs and tiRNAs (red dots) in tumor tissues compared with ATs. Fold change ≥ 1.5 and P -value < 0.05. (H) Differentially expressed tRFs and tiRNAs were separately calculated in PTC, and tRF-5c was the main down-regulated type (13/20). (I) Relative expression of tRF-30 in 30 pairs of tumor tissues and ATs was detected by qRT–PCR. (J) Relative expression of tRF-30 was detected by qRT–PCR in normal thyroid epithelial cells (Nthy-ori 3-1) and four PTC cell lines (BCPAP, TPC1, KTC1, and NPA87). Data were shown as mean ± SD. ( t test and Mann–Whitney U test, * P < 0.05, ** P < 0.01, and *** P < 0.001). Quantitative results were based on three independent experiments. Source data are available for this figure.

Article Snippet: The human PTC cell lines (BCPAP, TPC1, KTC1, NPA87) and the normal thyroid epithelial cells (Nthy-ori 3-1) were purchased from the iCell Bioscience Inc. All cell lines were identified by STR analysis and removed from mycoplasma contamination.

Techniques: Sequencing, Expressing, Quantitative RT-PCR, MANN-WHITNEY