Journal: Cancer biology & therapy

Article Title: Comparative analysis of the effects of a sphingosine kinase inhibitor to temozolomide and radiation treatment on glioblastoma cell lines.

doi: 10.1080/15384047.2017.1323583

Figure Lengend Snippet: Figure 4. (A) Western blot analysis for glutathione peroxidase 1 (GPx1, left) and O-6-methylguanine-DNA methyltransferase (MGMT, right) expressions in U87 and LN229 cells. LN18 cell lysates were used as a positive control for MGMT expression. (B) Cell death rescue experiment: clonogenic survival of U87 and LN229 cells after treatment with or without NAC in presence or absence of SKI (20 mM), TMZ (10 mM) and with or without X-ray irradiation (2 Gy). Non-treated cells were used as a control (CTRL). Data are presented as means and standard deviations. Values were normalized to the respective drug (SKI or TMZ) without irradiation. Statistical significance for differen- ces between samples with and without NAC was calculated using 2-way ANOVA and Bonferroni post-tests. p < 0.05; p < 0.01; (n D 3).

Article Snippet: The primary antibody against O-6-methylguanine-DNA methyltransferase (MGMT; Cell Signaling) was used at the dilution of 1:1000.

Techniques: Western Blot, Positive Control, Expressing, Irradiation, Control

Journal: Chemotherapy Research and Practice

Article Title: Inhibition of NF- κ B by Dehydroxymethylepoxyquinomicin Suppresses Invasion and Synergistically Potentiates Temozolomide and γ -Radiation Cytotoxicity in Glioblastoma Cells

doi: 10.1155/2013/593020

Figure Lengend Snippet: (a) The analysis of the methylation status of the MGMT gene in GBM cell lines revealed two groups: methylated (U87MG, U343MG-a, and LN319) and hemimethylated (U251, T98G and U138MG); however, only T98G and U138MG express this gene as detected by real time quantitative PCR (*a pool of 5 white matter samples was used as calibrator); (b) treatment with DHMEQ efficiently decreases the expression of MGMT after 24 h. Data represents two independent experiments in duplicate and are expressed as mean ± SEM; (c) comet assay showed that TMZ-induced DNA damage significantly increases in T98G and U138MG cells as a probable consequence of reduced MGMT expression after exposure to DHMEQ. Each value represents the mean derived from at least three individual experiments (mean ± SD).

Article Snippet: Real-time RT-PCR reactions were performed in triplicate in 10 μ L reactions using the inventoried TaqMan probes (Applied Biosystems, Foster City, CA, EUA) for BCL2 (Hs00608023_m1), BCL-XL (Hs00236329_m1), XIAP (Hs01597783_m1), MMP-2 (Hs01548727_m1), MMP-14 (Hs00237119_m1), uPA (Hs01547054_m1), TIMP-2 (Hs00234278_m1), and MGMT (Hs01037698_m1), on the ABI Prism 7500 Sequence Detector (Applied Biosystems, Foster City, CA, EUA).

Techniques: Methylation, Real-time Polymerase Chain Reaction, Expressing, Single Cell Gel Electrophoresis, Derivative Assay

Journal: Frontiers in Genetics

Article Title: Methylation associated with long- or short-term survival in glioblastoma patients from the Nordic phase 3 trial

doi: 10.3389/fgene.2022.934519

Figure Lengend Snippet: Patients from the Nordic trial included in the study. Patients with good prognostic factors were selected from each treatment arm (temozolomide -TMZ, radiotherapy 34Gy-RT 34Gy, and radiotherapy 60Gy- RT60Gy). Half of the tumors had methylated MGMT promoter (m-MGMT), and half had unmethylated MGMT promoter (u-MGMT). These patients were further divided into long-term survivors (LTS) and short-term survivors (STS). N, number of patients.

Article Snippet: The MGMT methylation status was analyzed with the MDxHealth method, Liège, Belgium, as mentioned in ).

Techniques: Methylation

Journal: Frontiers in Genetics

Article Title: Methylation associated with long- or short-term survival in glioblastoma patients from the Nordic phase 3 trial

doi: 10.3389/fgene.2022.934519

Figure Lengend Snippet: Patient characteristics.

Article Snippet: The MGMT methylation status was analyzed with the MDxHealth method, Liège, Belgium, as mentioned in ).

Techniques: Biomarker Discovery, Methylation

Journal: Frontiers in Genetics

Article Title: Methylation associated with long- or short-term survival in glioblastoma patients from the Nordic phase 3 trial

doi: 10.3389/fgene.2022.934519

Figure Lengend Snippet: Pie charts representing the structural genomic distribution of DMCs discovered in samples with long ( n = 3) and short ( n = 3) survival within the different treatment arms and with specified MGMT promoter methylation status. TSS200, 200 bases upstream transcription start site; TSS1500, 1,500 bases upstream transcription start site; UTR, untranslated region; IGR, intergenic region; 60Gy34Gy, combined RT.

Article Snippet: The MGMT methylation status was analyzed with the MDxHealth method, Liège, Belgium, as mentioned in ).

Techniques: Methylation

Journal: Neuro-Oncology Practice

Article Title: Correlation of commercially available quantitative MGMT (O-6-methylguanine-DNA methyltransferase) promoter methylation scores and GBM patient survival

doi: 10.1093/nop/npy028

Figure Lengend Snippet: Cox Regression Analysis of Overall Survival and Progression-Free Survival of 102 Primary Glioblastoma MGMT Unmethylated Patients With Substratification by MGMT Values

Article Snippet: Our patient cohort was derived retrospectively and consisted of newly diagnosed GBM patients seen at UCLA and KPLA between 2011 and 2016 with accessible LabCorp quantitative MGMT methylation values.

Techniques: Biomarker Discovery

Journal: Neuro-Oncology Practice

Article Title: Correlation of commercially available quantitative MGMT (O-6-methylguanine-DNA methyltransferase) promoter methylation scores and GBM patient survival

doi: 10.1093/nop/npy028

Figure Lengend Snippet: Kaplan-Meier analysis is used to A, compare the MGMT (1-1.99) vs MGMT (<1) and MGMT (≥2) patients. The MGMT (1-1.99) group’s median OS (25.4 months) falls in between the MGMT (≥2) (38.8 months) and MGMT (<1) (17.3 months) median OS values (Log-rank P = .001). B, PFS showed the same trend, namely the MGMT (1-1.99) group generated a higher median OS of 11.8 months compared to the MGMT (<1) group but lower than the MGMT (≥ 2) group, yielding 11.8 months vs 7.92 months and 18.0 months (Log-rank P < .0001), respectively. MGMT indicates O-6-methylguanine-DNA methyltransferase; mo, months; OS, overall survival; PFS, progression-free survival.

Article Snippet: Our patient cohort was derived retrospectively and consisted of newly diagnosed GBM patients seen at UCLA and KPLA between 2011 and 2016 with accessible LabCorp quantitative MGMT methylation values.

Techniques: Generated

Journal: Neuro-Oncology Practice

Article Title: Correlation of commercially available quantitative MGMT (O-6-methylguanine-DNA methyltransferase) promoter methylation scores and GBM patient survival

doi: 10.1093/nop/npy028

Figure Lengend Snippet: Cox Regression Analysis of Overall Survival and Progression-Free Survival of 165 Primary Glioblastoma Patients With Substratification by MGMT Values

Article Snippet: Our patient cohort was derived retrospectively and consisted of newly diagnosed GBM patients seen at UCLA and KPLA between 2011 and 2016 with accessible LabCorp quantitative MGMT methylation values.

Techniques: Biomarker Discovery

Journal: PLoS ONE

Article Title: Comprehensive Analysis of MGMT Promoter Methylation: Correlation with MGMT Expression and Clinical Response in GBM

doi: 10.1371/journal.pone.0016146

Figure Lengend Snippet: Each row represents an individual patient. For each patient the qMSP P positive result is shown as a black rectangle. The next column shows the results of the qMSP test (Labcorp). PFS is represented by a circle with a blackened circle representing greater than 1 year PFS. Positive MGMT protein expression (>15% tumor cells stained positive in IHC) is shown as black rounded rectangle and negative expression is shown as white rounded rectangle. MGMT mRNA expression is shown using color-coded rectangles. Absence of any of the shapes indicates results not available. All CpG sites belonging to the three regions, R1, R2 and R3 are shown within the blue outlined squares. The four distinct groups are outlined with red squares. The qMSP region is labeled with qMSP primers in R3. The transcription start site (TSS) of the MGMT gene is marked by an arrow.

Article Snippet: Real-time PCR of MGMT was performed on the ABI PRISM 7900 HT detection system using a taqman probe (Hs00172470_m1, Applied Biosystems) and taqman reagents under default conditions: 95°C for 10 minutes, 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute with human beta-glucuronidase (hGUS) as endogenous control.

Techniques: Expressing, Staining, Labeling

Journal: PLoS ONE

Article Title: Comprehensive Analysis of MGMT Promoter Methylation: Correlation with MGMT Expression and Clinical Response in GBM

doi: 10.1371/journal.pone.0016146

Figure Lengend Snippet: Significant individual CpG site correlations with MGMT mRNA expression, MGMT protein expression and PFS.

Article Snippet: Real-time PCR of MGMT was performed on the ABI PRISM 7900 HT detection system using a taqman probe (Hs00172470_m1, Applied Biosystems) and taqman reagents under default conditions: 95°C for 10 minutes, 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute with human beta-glucuronidase (hGUS) as endogenous control.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Comprehensive Analysis of MGMT Promoter Methylation: Correlation with MGMT Expression and Clinical Response in GBM

doi: 10.1371/journal.pone.0016146

Figure Lengend Snippet: All three regions of the MGMT promoter for each patient are classified as methylated (M) or unmethylated (UM). Based on the status of these three regions, patients are classified into four major groups: Group A) all regions unmethylated, Group B) one region methylated, Group C) two regions methylated, and Group D) all three regions methylated. For correlation analysis, Groups B and C were further subdivided as indicated.

Article Snippet: Real-time PCR of MGMT was performed on the ABI PRISM 7900 HT detection system using a taqman probe (Hs00172470_m1, Applied Biosystems) and taqman reagents under default conditions: 95°C for 10 minutes, 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute with human beta-glucuronidase (hGUS) as endogenous control.

Techniques: Methylation

Journal: PLoS ONE

Article Title: Comprehensive Analysis of MGMT Promoter Methylation: Correlation with MGMT Expression and Clinical Response in GBM

doi: 10.1371/journal.pone.0016146

Figure Lengend Snippet: Correlation of MGMT promoter methylation patterns with MGMT mRNA expression, MGMT protein expression and PFS.

Article Snippet: Real-time PCR of MGMT was performed on the ABI PRISM 7900 HT detection system using a taqman probe (Hs00172470_m1, Applied Biosystems) and taqman reagents under default conditions: 95°C for 10 minutes, 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute with human beta-glucuronidase (hGUS) as endogenous control.

Techniques: Methylation, Expressing

Journal: PLoS ONE

Article Title: Comprehensive Analysis of MGMT Promoter Methylation: Correlation with MGMT Expression and Clinical Response in GBM

doi: 10.1371/journal.pone.0016146

Figure Lengend Snippet: Kaplan-Meier estimation of PFS according to MGMT promoter methylation status determined using: A) 3R classification method - The median PFS for the methylated group was 540 days vs. 210 days for the unmethylated group (HR = 5.23, 95% CI [2.089–13.097], p<0.0001), B) qMSP P - The median PFS for the methylated group was 373 days vs. 224 days for the unmethylated group (HR = 1.707, 95% CI [0.728–4.003], p = 0.213), and C) MS-MLPA - The median PFS for the methylated group was 540 days vs. 210 days for the unmethylated group (HR = 3.076, 95% CI [1.301–7.27], p = 0.007).

Article Snippet: Real-time PCR of MGMT was performed on the ABI PRISM 7900 HT detection system using a taqman probe (Hs00172470_m1, Applied Biosystems) and taqman reagents under default conditions: 95°C for 10 minutes, 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute with human beta-glucuronidase (hGUS) as endogenous control.

Techniques: Methylation

Journal: PLoS ONE

Article Title: Comprehensive Analysis of MGMT Promoter Methylation: Correlation with MGMT Expression and Clinical Response in GBM

doi: 10.1371/journal.pone.0016146

Figure Lengend Snippet: Kaplan-Meier estimation of PFS according to MGMT promoter methylation status determined using: A. MS-MLPA (p-value = 0.002) and B. qMSP (p-value = 0.089).

Article Snippet: Real-time PCR of MGMT was performed on the ABI PRISM 7900 HT detection system using a taqman probe (Hs00172470_m1, Applied Biosystems) and taqman reagents under default conditions: 95°C for 10 minutes, 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute with human beta-glucuronidase (hGUS) as endogenous control.

Techniques: Methylation