Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epigenetic and post-transcriptional regulation of CD16 expression during human natural killer cell development 1

doi: 10.4049/jimmunol.1701128

Figure Lengend Snippet: DNA methylation and isoform-specific expression within the promoter regions of FCGR3A and FCGR3B. A) Representation of the FCGR3 locus spanning approximately 100 kb of chromosome 1q23.3 displaying transcripts annotated in the RefSeq database (adapted from the UCSC Genome Browser). Below, promoter regions of FCGR3A and FCGR3B are enlarged displaying the various transcript variants annotated in the RefSeq database that differ in the 5′ region. MassARRAY amplicons (green bars) used are shown along with the positions of CpG dinucleotides (green markers). TSS, transcriptional start site. B) The MassARRAY EpiTYPER assay was used to interrogate the DNA methylation levels of CpGs across the promoter region of FCGR3A and FCGR3B in a gene-specific manner in sorted NK CD16a+ and CD16a- fractions. Methylation plots are aligned to enlargements of gene promoters (in B) according to the hg19 genome assembly. C, D) DNA methylation levels in NK cell lines and neutrophils, respectively. E) The expression ratio of FCGR3A to FCGR3B in CD16a+ NK cells and neutrophils as determined by the MassARRAY iPLEX assay. F) Quantitative RT-PCR analysis of 5′ transcript variants using primers specific for variant 1 or variants 3 and 2 indicating promoter usage in NK cells, neutrophils, and NK cell lines YT and NKL (both primer pairs do not distinguish FCGR3A vs. FCGR3B). D1-4 represents donor numbers 1–4. Expression values expressed relative to the average of four housekeeping (HK) genes.

Article Snippet: Antibodies and flow cytometric analysis The following antibodies were used to stain human peripheral blood cells: CD3 (SK7, BD Biosciences), CD14 (TÜK4, Miltenyi), CD15 (VIMC6, Miltenyi), CD16 (VEP13, Miltenyi), CD16 (3G8, BD Biosciences), and CD56 (N901, Beckman Coulter).

Techniques: DNA Methylation Assay, Expressing, MassARRAY EpiTYPER assay, Methylation, Quantitative RT-PCR, Variant Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epigenetic and post-transcriptional regulation of CD16 expression during human natural killer cell development 1

doi: 10.4049/jimmunol.1701128

Figure Lengend Snippet: Analysis of DNA methylation- and lineage-specific activity of FCGR3 promoter sequences. A) Illustration of FCGR3A and FCGR3B sequences cloned into luciferase constructs (black bars). B) Four separate promoter sequence fragments were cloned from either FCGR3A (left) or FCGR3B (right) and transfected in various cell lines. Luciferase assays showing sequence- and gene-specific activity (relative to empty-vector control). Pmed1-A and Pmed1-B were additionally methylated in vitro prior to transfection. Error bars represent SEM of n=3 individual experiments; ND, not done. C) Sequence alignment of Pmed1-A and Pmed1-B with numbered CpGs. All sequence variants are enlarged below; an 8 bp repeat occurring in the vicinity of CpG#6 is highlighted in blue, asterisks below the sequence indicate homology. D) Luciferase activity following site-directed mutagenesis of CpG#s 1&2 of Pmed1-A compared to unaltered FCGR3A and FCGR3B sequences in YT and K562 cells.

Article Snippet: Antibodies and flow cytometric analysis The following antibodies were used to stain human peripheral blood cells: CD3 (SK7, BD Biosciences), CD14 (TÜK4, Miltenyi), CD15 (VIMC6, Miltenyi), CD16 (VEP13, Miltenyi), CD16 (3G8, BD Biosciences), and CD56 (N901, Beckman Coulter).

Techniques: DNA Methylation Assay, Activity Assay, Clone Assay, Luciferase, Construct, Sequencing, Transfection, Plasmid Preparation, Control, Methylation, In Vitro, Mutagenesis

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epigenetic and post-transcriptional regulation of CD16 expression during human natural killer cell development 1

doi: 10.4049/jimmunol.1701128

Figure Lengend Snippet: Identification of miR-218 as a potential regulator of FCGR3A. (A) Expression ratio of predicted miRNAs that were present in the miRNA expression array comparing CD16a+ and CD16a- NK cells freshly isolated from adult peripheral blood. A ratio <1 indicates low expression in CD16a+ NK cells while a ratio >1 indicates high expression in CD16a+ NK cells. (B) Predicted miRNA regulators of FCGR3A have putative binding sites in the FCGR3A 3′ UTR. (C) Validation of expression of predicted miRNA regulators of FCGR3A by qPCR (n=3). (D) Luciferase expression as a ratio of firefly/renilla for each expression vector (n=2). Data are presented as mean±SD, * indicates p<0.05.

Article Snippet: Antibodies and flow cytometric analysis The following antibodies were used to stain human peripheral blood cells: CD3 (SK7, BD Biosciences), CD14 (TÜK4, Miltenyi), CD15 (VIMC6, Miltenyi), CD16 (VEP13, Miltenyi), CD16 (3G8, BD Biosciences), and CD56 (N901, Beckman Coulter).

Techniques: Expressing, Isolation, Binding Assay, Biomarker Discovery, Luciferase, Plasmid Preparation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epigenetic and post-transcriptional regulation of CD16 expression during human natural killer cell development 1

doi: 10.4049/jimmunol.1701128

Figure Lengend Snippet: MiR-218 negatively regulates CD16a in primary human NK cells. Primary human NK cells were enriched by magnetic selection to >70% purity and infected with lentivirus containing either miR-218 or empty vector. 48hr after infection, NK cells were sorted as live/CD3-/CD56+/GFP+ lymphocytes. (A) Representative histogram plot (of one of six donors) of CD16a expression in live/CD3-/CD56+/GFP+ primary human NK cells infected with miR-218 or empty vector virus. (B) CD16a expression in primary human NK cells infected with miR-218 or empty vector virus (* indicates p=0.05). (C) Validation of miR-218 over-expression by real-time PCR (** indicates p<0.01). (D) FCGR3A mRNA expression assessed by real-time RT-PCR in sorted NK cells infected with either miR-218 or empty vector (*** indicates p<0.005). (B–D) Data are presented as mean±SD, n=6.

Article Snippet: Antibodies and flow cytometric analysis The following antibodies were used to stain human peripheral blood cells: CD3 (SK7, BD Biosciences), CD14 (TÜK4, Miltenyi), CD15 (VIMC6, Miltenyi), CD16 (VEP13, Miltenyi), CD16 (3G8, BD Biosciences), and CD56 (N901, Beckman Coulter).

Techniques: Selection, Infection, Plasmid Preparation, Expressing, Virus, Biomarker Discovery, Over Expression, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: DEGs in non-metastatic vs. metastatic samples from GEO database. (A,B) Analysis of DEGs between 17 noninvasive BAC and 23 AC-mixed subtype invasive lung adenocarcinomas (GSE27716). A total of 2,172 DEGs, namely, 2,077 downregulated and 95 upregulated genes, were obtained (|logFC|>0.4; adj.P.value<0.01); (C,D) Analysis of DEGs in A549 cells before and after TGFβ1 induced EMT (GSE49644). A total of 2,513 DEGs, namely, 1,506 downregulated genes and 1,007 upregulated genes, were obtained (|logFC|>0.8; adj.P.value<0.001); (E,F) Analysis of DEGs in H358 cells with or without TGFβ1 treatment (GSE49644). A total of 2,295 DEGs, namely, 1,412 downregulated genes and 883 upregulated genes, were obtained (|logFC|>0.8; adj.P.value<0.001). The analysis results of DEGs are presented as heatmaps and volcano plots; (G,H) Venn diagrams showing upregulated (H) and downregulated (G) genes whose dysregulated expression pattern was shared by two GEO microarray database; (I) Analysis of DEGs between HBE-Ctrl and HBE-5Aza cells. HBE cells were treated with 5-Aza (10 μmol/L) for 96 h, and the medium was exchanged every day; (J) Venn diagrams show overlapping genes shared by 506 upregulated genes induced by 5Aza and 18 metastatic-associated downregulated genes. DEG; differentially expressed genes; GEO; Gene Expression Omnibus; BAC; bronchioloalveolar carcinomas; AC, adenocarcinomas; TGFβ1, transforming growth factor β1; EMT, epithelial-to-mesenchymal transition; HBE, human bronchial epithelial.

Article Snippet: Recombinant human TGFβ1 protein (R&D systems, Minneapolis, USA, #240-B-002) was prepared as a stock solution of 20 μg/mL in sterile 4 mmol/L HCl containing 1 mg/mL bovine serum albumin.

Techniques: Expressing, Microarray, Gene Expression

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: Stomatin is essential for transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT) in A549 cells. (A) Representative images of TGFβ1-induced EMT in A549 cells. Scale bars represent 100 μm; (B) A549 cells were treated with 5 ng/mL TGFβ1 for indicated times, and the expressions of stomatin, E-cadherin, Vimentin, N-cadherin and Slug were assessed by western blotting. β-actin was used as a loading control; (C,D) Immunofluorescences were performed to demonstrate the inhibition of stomatin by TGFβ1. E-cadherin (C) and Vimentin (D) staining indicated the induction of EMT in A549 by TGFβ1. Scale bars represent 30 μm. (E) Semiquantitative scoring was performed on stomatin and TGFβ1 staining in lung adenocarcinoma samples. Stomatin was negatively correlated with TGFβ1 in stage III (r 2 =−0.501, P=0.048) tumors; (F) A549-NC and A549-STOM cells were exposed to TGFβ1 (5 ng/mL) for 48 h. The expression levels of N-cadherin, E-cadherin, and stomatin were analyzed by western blotting. β-actin was used as a loading control.

Article Snippet: Recombinant human TGFβ1 protein (R&D systems, Minneapolis, USA, #240-B-002) was prepared as a stock solution of 20 μg/mL in sterile 4 mmol/L HCl containing 1 mg/mL bovine serum albumin.

Techniques: Western Blot, Control, Inhibition, Staining, Expressing

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: Stomatin is inhibited during transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT) in Panc-1 and MDA-MB-231 cells. (A) Panc-1 cells were treated with 10 ng/mL TGFβ1 for the indicated times, and expressions of stomatin, E-cadherin, and N-cadherin were assessed by western blotting. β-actin was used as a loading control; (B,C) Immunofluorescence microscopy was performed to demonstrate the inhibition of stomatin expression by TGFβ1. E-cadherin (B) and Vimentin (C) staining indicated the induction of EMT in Panc-1 by TGFβ1. Scale bars represents 30 μm; (D) MDA-MB-231 cells were treated with 10 ng/mL TGFβ1 for 2 d, and the expression of stomatin was assessed by real-time polymerase chain reaction (PCR). *, P<0.05. **, P<0.001. Three independent experiments were performed; (E) Semiquantitative scoring was performed, and stomatin scores had no correlation with TGFβ1 scores in stage I and II (r=0.1, P=0.477, n=53) lung adenocarcinomas.

Article Snippet: Recombinant human TGFβ1 protein (R&D systems, Minneapolis, USA, #240-B-002) was prepared as a stock solution of 20 μg/mL in sterile 4 mmol/L HCl containing 1 mg/mL bovine serum albumin.

Techniques: Western Blot, Control, Immunofluorescence, Microscopy, Inhibition, Expressing, Staining, Real-time Polymerase Chain Reaction

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: Mechanisms of stomatin inhibition during transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT). (A) A549 cells were treated with or without 5 ng/mL TGFβ1. After 12 h, cells were treated with 5 μg/mL cycloheximide (CHX) for the indicated times with or without TGFβ1. Endogenous stomatin was detected by western blotting. β-actin was used as a loading control; (B) A549 cells were stimulated with 5 ng/mL TGFβ1 for 12 h before 10 μg/mL actinomycin D (AcD) treatment; the cells were incubated with AcD for the indicated times. The expression of stomatin was determined by quantitative real-time polymerase chain reaction (PCR). Three independent experiments were performed; (C) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d with or without 5-Aza. Expressions of E-cadherin, and stomatin were analyzed by real-time PCR. *, P<0.05, **, P<0.001. Three independent experiments were performed.

Article Snippet: Recombinant human TGFβ1 protein (R&D systems, Minneapolis, USA, #240-B-002) was prepared as a stock solution of 20 μg/mL in sterile 4 mmol/L HCl containing 1 mg/mL bovine serum albumin.

Techniques: Inhibition, Western Blot, Control, Incubation, Expressing, Real-time Polymerase Chain Reaction

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through methylation-dependent transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom MassARRAY assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).

Article Snippet: Recombinant human TGFβ1 protein (R&D systems, Minneapolis, USA, #240-B-002) was prepared as a stock solution of 20 μg/mL in sterile 4 mmol/L HCl containing 1 mg/mL bovine serum albumin.

Techniques: Methylation, Real-time Polymerase Chain Reaction, Microarray, Sequenom Massarray Assay, Two Tailed Test, Expressing, Western Blot, Control

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: Stomatin is essential for transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT) in A549 cells. (A) Representative images of TGFβ1-induced EMT in A549 cells. Scale bars represent 100 μm; (B) A549 cells were treated with 5 ng/mL TGFβ1 for indicated times, and the expressions of stomatin, E-cadherin, Vimentin, N-cadherin and Slug were assessed by western blotting. β-actin was used as a loading control; (C,D) Immunofluorescences were performed to demonstrate the inhibition of stomatin by TGFβ1. E-cadherin (C) and Vimentin (D) staining indicated the induction of EMT in A549 by TGFβ1. Scale bars represent 30 μm. (E) Semiquantitative scoring was performed on stomatin and TGFβ1 staining in lung adenocarcinoma samples. Stomatin was negatively correlated with TGFβ1 in stage III (r 2 =−0.501, P=0.048) tumors; (F) A549-NC and A549-STOM cells were exposed to TGFβ1 (5 ng/mL) for 48 h. The expression levels of N-cadherin, E-cadherin, and stomatin were analyzed by western blotting. β-actin was used as a loading control.

Article Snippet: The lung cancer tissue microarray slides included 69 cases of LUAD with detailed information of each patient were purchased from Shanghai Outdo Biotech Co., Ltd. Immunohistochemistry of these slides was performed using antibodies, such as TGFβ1 (abcam, #ab215715) and stomatin (Proteintech, Rosemont, USA, #12046-1-AP).

Techniques: Western Blot, Control, Inhibition, Staining, Expressing

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: Stomatin is inhibited during transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT) in Panc-1 and MDA-MB-231 cells. (A) Panc-1 cells were treated with 10 ng/mL TGFβ1 for the indicated times, and expressions of stomatin, E-cadherin, and N-cadherin were assessed by western blotting. β-actin was used as a loading control; (B,C) Immunofluorescence microscopy was performed to demonstrate the inhibition of stomatin expression by TGFβ1. E-cadherin (B) and Vimentin (C) staining indicated the induction of EMT in Panc-1 by TGFβ1. Scale bars represents 30 μm; (D) MDA-MB-231 cells were treated with 10 ng/mL TGFβ1 for 2 d, and the expression of stomatin was assessed by real-time polymerase chain reaction (PCR). *, P<0.05. **, P<0.001. Three independent experiments were performed; (E) Semiquantitative scoring was performed, and stomatin scores had no correlation with TGFβ1 scores in stage I and II (r=0.1, P=0.477, n=53) lung adenocarcinomas.

Article Snippet: The lung cancer tissue microarray slides included 69 cases of LUAD with detailed information of each patient were purchased from Shanghai Outdo Biotech Co., Ltd. Immunohistochemistry of these slides was performed using antibodies, such as TGFβ1 (abcam, #ab215715) and stomatin (Proteintech, Rosemont, USA, #12046-1-AP).

Techniques: Western Blot, Control, Immunofluorescence, Microscopy, Inhibition, Expressing, Staining, Real-time Polymerase Chain Reaction

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: Mechanisms of stomatin inhibition during transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT). (A) A549 cells were treated with or without 5 ng/mL TGFβ1. After 12 h, cells were treated with 5 μg/mL cycloheximide (CHX) for the indicated times with or without TGFβ1. Endogenous stomatin was detected by western blotting. β-actin was used as a loading control; (B) A549 cells were stimulated with 5 ng/mL TGFβ1 for 12 h before 10 μg/mL actinomycin D (AcD) treatment; the cells were incubated with AcD for the indicated times. The expression of stomatin was determined by quantitative real-time polymerase chain reaction (PCR). Three independent experiments were performed; (C) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d with or without 5-Aza. Expressions of E-cadherin, and stomatin were analyzed by real-time PCR. *, P<0.05, **, P<0.001. Three independent experiments were performed.

Article Snippet: The lung cancer tissue microarray slides included 69 cases of LUAD with detailed information of each patient were purchased from Shanghai Outdo Biotech Co., Ltd. Immunohistochemistry of these slides was performed using antibodies, such as TGFβ1 (abcam, #ab215715) and stomatin (Proteintech, Rosemont, USA, #12046-1-AP).

Techniques: Inhibition, Western Blot, Control, Incubation, Expressing, Real-time Polymerase Chain Reaction

Journal: Chinese Journal of Cancer Research

Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

doi: 10.21147/j.issn.1000-9604.2019.06.09

Figure Lengend Snippet: During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through methylation-dependent transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom MassARRAY assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).

Article Snippet: The lung cancer tissue microarray slides included 69 cases of LUAD with detailed information of each patient were purchased from Shanghai Outdo Biotech Co., Ltd. Immunohistochemistry of these slides was performed using antibodies, such as TGFβ1 (abcam, #ab215715) and stomatin (Proteintech, Rosemont, USA, #12046-1-AP).

Techniques: Methylation, Real-time Polymerase Chain Reaction, Microarray, Sequenom Massarray Assay, Two Tailed Test, Expressing, Western Blot, Control

Journal: Oncology Reports

Article Title: Prognostic implication of PTPRH hypomethylation in non-small cell lung cancer

doi: 10.3892/or.2015.4082

Figure Lengend Snippet: CpG sites analyzed by the MassARRAY system.

Article Snippet: DNA methylation levels obtained by the MassARRAY system and Infinium assay correlated well (r=0.952, P=1.44×10 −73 ), confirming the reliability of the latter assay.

Techniques:

Journal: Oncology Reports

Article Title: Prognostic implication of PTPRH hypomethylation in non-small cell lung cancer

doi: 10.3892/or.2015.4082

Figure Lengend Snippet: (A) Median DNA methylation levels of PTPRH in N and T with no recurrence and those with recurrence in LC-C2. DNA methylation levels of each CpG unit were evaluated quantitatively using the MassARRAY system. The error bars are defined by 25/75% quartiles. DNA methylation levels at most CpG sites showed a significant decrease in tumorous tissue with recurrence compared with those with no recurrence. Significantly different methylated CpG sites are marked as * P<0.05 and ** P<0.01). (B) Kaplan-Meier survival curves of patients with PTPRH hypermethylation (>0.782 at CpG_9.10) and hypomethylation (≤0.782). The cutoff was determined by ROC curve analysis. The recurrence-free (P=1.64×10 −4 ) and overall (P=5.54×10 −5 ) survival rates of patients with hypomethylation were significantly lower compared to those of patients with hypermethylation (log-rank test). N, non-cancerous lung tissue; T, corresponding tumorous tissue; ROC, receiver operating characteristic.

Article Snippet: DNA methylation levels obtained by the MassARRAY system and Infinium assay correlated well (r=0.952, P=1.44×10 −73 ), confirming the reliability of the latter assay.

Techniques: DNA Methylation Assay, Methylation