scholarly journals MGMT Gene Promoter Methylation Status – Assessment of Two Pyrosequencing Kits and Three Methylation-specific PCR Methods for their Predictive Capacity in Glioblastomas

2018 ◽  
Vol 15 (6) ◽  
pp. 437-446 ◽  
Author(s):  
LENE E. JOHANNESSEN ◽  
PETTER BRANDAL ◽  
TOR ÅGE MYKLEBUST ◽  
SVERRE HEIM ◽  
FRANCESCA MICCI ◽  
...  
Keyword(s):  
Promoter Methylation ◽  
Methylation Status ◽  
Gene Promoter ◽  
Predictive Capacity ◽  
Promoter Methylation Status ◽  
Mgmt Gene ◽  
Methylation Specific Pcr ◽  
Specific Pcr ◽  
Status Assessment

scholarly journals Influence of Intratumor Heterogeneity on the Predictivity of MGMT Gene Promoter Methylation Status in Glioblastoma

2020 ◽  
Vol 10 ◽  
Author(s):  
Giovanni Brigliadori ◽  
Giulia Goffredo ◽  
Daniela Bartolini ◽  
Luigino Tosatto ◽  
Lorena Gurrieri ◽  
...  
Keyword(s):  
Promoter Methylation ◽  
Methylation Status ◽  
Gene Promoter ◽  
Intratumor Heterogeneity ◽  
Promoter Methylation Status ◽  
Mgmt Gene

INI1 immunohistochemical expression in glioblastoma: correlation with MGMT gene promoter methylation status and patient survival

Pathology ◽  
2011 ◽  
Vol 43 (1) ◽  
pp. 17-23 ◽  
Author(s):  
Elena Zunarelli ◽  
Nazzarena Bigiani ◽  
Giuliana Sartori ◽  
Mario Migaldi ◽  
Alessandro Sgambato ◽  
...  
Keyword(s):  
Promoter Methylation ◽  
Patient Survival ◽  
Methylation Status ◽  
Gene Promoter ◽  
Immunohistochemical Expression ◽  
Promoter Methylation Status ◽  
Mgmt Gene

scholarly journals MS-FLAG, a Novel Real-Time Signal Generation Method for Methylation-Specific PCR

2007 ◽  
Vol 53 (12) ◽  
pp. 2119-2127 ◽  
Author(s):  
Cinzia Bonanno ◽  
Erlet Shehi ◽  
Daniel Adlerstein ◽  
G Mike Makrigiorgos
Keyword(s):  
Real Time ◽  
Promoter Methylation ◽  
Real Time Pcr ◽  
Bisulfite Sequencing ◽  
Methylation Status ◽  
Gene Promoter ◽  
Signal Generation ◽  
Time Signal ◽  
Specific Pcr ◽  
Generation Technology

Abstract Background: Aberrant promoter methylation is a major mechanism for silencing tumor suppressor genes in cancer. Detection of hypermethylation is used as a molecular marker for early cancer diagnosis, as a prognostic index, or to define therapeutic targets for reversion of aberrant methylation. We report on a novel signal generation technology for real-time PCR to detect gene promoter methylation. Methods: FLAG (fluorescent amplicon generation) is a homogeneous signal generation technology based on the exceptionally thermostable endonuclease PspGI. FLAG provides real-time signal generation during PCR by PspGI-mediated cleavage of quenched fluorophores at the 5′ end of double-stranded PCR products. Methylation-specific PCR (MSP) applied on bisulfite-treated DNA was adapted to a real-time format (methylation-specific FLAG; MS-FLAG) for quantifying methylation in the promoter of CDKN2A (p16), GATA5, and RASSF1. We validated MS-FLAG on plasmids and genomic DNA with known methylation status and applied it to detection of methylation in a limited number of clinical samples. We also conducted bisulfite sequencing on these samples. Results: Real-time PCR results obtained via MS-FLAG agreed with results obtained via conventional, gel-based MSP. The new technology showed high specificity, sensitivity (2–3 plasmid copies), and selectivity (0.01% of methylated DNA) on control samples. It enabled correct prediction of the methylation status of all 3 gene promoters in 21 lung adenocarcinoma samples, as confirmed by bisulfite sequencing. We also developed a multiplex MS-FLAG assay for GATA5 and RASSF1 promoters. Conclusion: MS-FLAG provides a new, quantitative, high-throughput method for detecting gene promoter methylation and is a convenient alternative to agarose gel-based MSP for screening methylation. In addition to methylation, FLAG-based real-time signal generation may have broad applications in DNA diagnostics.

Protein Expression of p15 and p21 Plays an Unfavorable Prognostic Role in Adult Acute Lymphoblastic Leukemia (ALL) Patients Independently of Their Gene Promoter Methylation Status.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2802-2802
Author(s):  
Fabiana De Cave ◽  
Maria Teresa Petrucci ◽  
Chiara Gregorj ◽  
Maria Rosaria Ricciardi ◽  
Samantha Decandia ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Protein Expression ◽  
Promoter Methylation ◽  
Lymphoblastic Leukemia ◽  
Methylation Status ◽  
Gene Promoter ◽  
Prognostic Role ◽  
P21 Protein ◽  
Promoter Methylation Status ◽  
P16 Protein

Abstract Epigenetic silencing of tumor suppressor (TS) genes is a hallmark in human leukemias, particularly through DNA methylation. Cyclin-dependent kinase inhibitors (CKI) are, among other genes, frequently found methylated in their promoter region. This epigenetic modification has been described also in acute lymphoblastic leukemia (ALL). However, the relationship between aberrant DNA methylation and protein expression of TS genes has not yet been extensively evaluated in adult ALL series. The aim of this study was to analyze in primary cells from newly diagnosed adult ALL patients, uniformly treated according to the LAL2000 GIMEMA protocol, the promoter methylation status of p73, p21, p15 and p16, evaluating in addition the p21, p15 and p16 protein expression. The DNA methylation status of promoter regions was investigated, according to cell availability, using a widely accepted method based on bisulfite modification of DNA, followed by methylation-specific PCR assay (MSP). Protein expression was evaluated by Western blot. Normal peripheral blood lymphocytes, as already described, resulted unmethylated for p73, p21, p15 and p16, and did not express the p21, p15 and p16 proteins. In ALL patients, in contrast, only the p21 promoter region was found constantly unmethylated. The p15, p16 and p73 promoter genes were found methylated in 15/37 (40.5%), 8/43 (18.6%) and 9/36 (25%) patients, respectively. Only 2/23 cases (8.6%) resulted simultaneously methylated for p15, p16 and p73. The p21 and p15 protein expression was found in 28/85 (32.9%) and 44/85 cases (51.8%), respectively. The p16 protein, in contrast, was never expressed. The p16 methylation was associated with the T-ALL (P=0.005) phenotype and with higher white blood cell (WBC) counts (P=0.027). Resistance to spontaneous induction of apoptosis was significantly associated with p21 protein expression (P=0.019) and its co-expression with p15 (P=0.049). Achievement of CR was not influenced by gene methylation status, nor by single protein expression. Interestingly, the co-expression of p15 and p21 was associated with failure to induction treatment: only 6/63 (9.5%) patients co-expressing p15 and p21 obtained a CR (P=0.027). Multivariate analysis confirmed the unfavorable role of this protein co-expression (P=0.059) on CR achievement. In contrast, once patients achieved remission, p21 protein expression was associated with a prolonged DFS, as confirmed by multivariate analysis for DFS (P=0.039). In conclusion, p15 and p21 protein expression plays an unfavorable prognostic role in adult ALL patients independently of the p73, p21, p15 and p16 gene promoter methylation status.

Defining the cutoff value of MGMT gene promoter methylation and its predictive capacity in glioblastoma

2016 ◽  
Vol 128 (2) ◽  
pp. 333-339 ◽  
Author(s):  
Giovanni Brigliadori ◽  
Flavia Foca ◽  
Monia Dall’Agata ◽  
Claudia Rengucci ◽  
Elisabetta Melegari ◽  
...  
Keyword(s):  
Promoter Methylation ◽  
Gene Promoter ◽  
Predictive Capacity ◽  
Cutoff Value ◽  
Mgmt Gene

TLR2 and TLR4 gene promoter methylation status during chronic periodontitis

2011 ◽  
Vol 38 (11) ◽  
pp. 975-983 ◽  
Author(s):  
Naila Francis Paulo De Oliveira ◽  
Denise Carleto Andia ◽  
Aline Cristiane Planello ◽  
Silvana Pasetto ◽  
Marcelo Rocha Marques ◽  
...  
Keyword(s):  
Promoter Methylation ◽  
Chronic Periodontitis ◽  
Methylation Status ◽  
Gene Promoter ◽  
Promoter Methylation Status ◽  
Tlr4 Gene
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