2016
DOI: 10.1186/s13148-016-0204-7
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MGMT promoter methylation determined by HRM in comparison to MSP and pyrosequencing for predicting high-grade glioma response

Abstract: BackgroundThe DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT) causes resistance of cancer cells to alkylating agents and, therefore, is a well-established predictive marker for high-grade gliomas that are routinely treated with alkylating drugs. Since MGMT is highly epigenetically regulated, the MGMT promoter methylation status is taken as an indicator of MGMT silencing, predicting the outcome of glioma therapy. MGMT promoter methylation is usually determined by methylation specific PCR (MSP),… Show more

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Cited by 60 publications
(54 citation statements)
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“…Thus, repeated TMZ treatment of MGMT lacking U87 cells gave rise to an up to 200-fold increase in the MGMT mRNA level ( Figure S5a-c). Concomitantly, analyzing promoter methylation by HRM (Switzeny et al 2016), a reduced methylation of the promoter was found after repeated TMZ treatment cycles. This again suggests that chronic TMZ treatment leads to selection of cells harboring a weak methylated promoter or to re-activation of the MGMT gene.…”
Section: Discussionmentioning
confidence: 99%
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“…Thus, repeated TMZ treatment of MGMT lacking U87 cells gave rise to an up to 200-fold increase in the MGMT mRNA level ( Figure S5a-c). Concomitantly, analyzing promoter methylation by HRM (Switzeny et al 2016), a reduced methylation of the promoter was found after repeated TMZ treatment cycles. This again suggests that chronic TMZ treatment leads to selection of cells harboring a weak methylated promoter or to re-activation of the MGMT gene.…”
Section: Discussionmentioning
confidence: 99%
“…Promoter methylation-specific PCR of 17 glioblastoma cell lines was performed as described previously (Christmann et al 2010). Quantitative high resolution melt (HRM) analysis was performed as described (Switzeny et al 2016).…”
Section: Preparation Of Rna Semi-quantitative Pcr and Rt Qpcrmentioning
confidence: 99%
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“…This work reports an immuno‐DNA‐based method for the fast assessment of specific methylations at MGMT tumor suppressor gene promoter, considered a useful predictor of therapy responsiveness in patients with glioblastoma . The designed biosensing strategy is shown schematically in Figure a. HOOC‐MBs were modified with a specific 5‐methylcytosine (anti‐5‐mC) monoclonal antibody, through covalent immobilization by EDC/Sulfo‐NHS chemistry, to capture any 5‐mC‐methylated ssDNA sequence.…”
Section: Figurementioning
confidence: 99%