Association between MGMT Enhancer Methylation and MGMT Promoter Methylation, MGMT Protein Expression, and Overall Survival in Glioblastoma
Abstract
:1. Introduction
2. Materials and Methods
2.1. Samples and Cell Culturing
2.2. DNA Extraction and Bisulfite Conversion
2.3. Primer Design and PCR Conditions for DNA Methylation Analysis
2.4. PSQ of PCR Products
2.5. Data Analysis and Statistics
3. Results
3.1. MGMT Promoter and Enhancer Methylation
3.1.1. Promoter Methylation
3.1.2. Enhancer 1 Methylation
3.1.3. Enhancer 2 Methylation
3.1.4. Enhancer 3 Methylation
3.1.5. Enhancer 4 Methylation
3.2. Association between the MGMT Promoter and/or Enhancer Methylation
3.3. Association of the MGMT Promoter and Enhancer Methylation with MGMT Protein Expression
3.4. Association between the MGMT Promoter and Enhancer Methylation and Overall Survival
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Region | Primer Set | Primer Sequence (5′→3′) | Length [bp] | CpGs Analyzed |
---|---|---|---|---|
enhancer 1 | A | F: AGAATGTAGATTTGGGATTAGTTAAT | 229 | 12–19 |
(hs737) | R: [Btn] TAAAATACAAAATATACCCTCCAACA | |||
27 CpGs | S: TATATAAAGAAGGTTGGT | |||
enhancer 2 | A | F: AGTTAGGAAATTAGAAATGGAATGTTT | 255 | 05–08 |
(Chen et al.) | R: [Btn] CAAATCACACTCTAAATTCCCAATT | |||
46 CpGs | S: TGGTATTAGAGGTTA | |||
B | F: [Btn] TTAAATAAGTGGTTTAGGTAGAGG | 137 | 11–18 | |
R: TACTAAACATTCCATTTCTAATTTCC | ||||
S: CCATTTCTAATTTCCTAACTC | ||||
C | F: GTTGTAGGGTATATGAGTTTAGAT | 271 | 24–27 | |
R: [Btn] TTCATAACTCAAATTAACACACACT | ||||
S: TTTTGTGTTGAATGG | ||||
D | F: GAGGTTATTTGGAAAGTTGAGAT | 286 | 37–39 | |
R: [Btn] CTAATAATCCAAACCCTCTATTC | ||||
S: TTTAGTGTTATGGGAG | ||||
enhancer 3 | A | F: TGTGTTAGTTTTAGTGGTTTAGA | 138 | 15–22 |
(hs699) | R: [Btn] TAACACACAAACCAATCTCTC | |||
33 CpGs | S: TAGTTTTAGTGGTTTAGAAGT | |||
enhancer 4 | A | F: [Btn] GGAATGTGTTATTTAATTGGTATGT | 204 | 01–03 |
(hs696) | R: CAAATCCCACAACAAATCCTTAT | |||
26 CpGs | S: TCAAAAAAAAAAAATCACC | |||
B | F: GAGGTTTGATATAAGTAATGATGG | 131 | 07–08 | |
R: [Btn] CCTCCTAATCCCACAATACAA | ||||
S: TAAGTAATGATGGTATG | ||||
C | F: AGGTTTGATATAAGTAATGATGGTAT | 257 | 09–13 | |
R: [Btn] CRTATTCTCTCCCACTTCAATA | ||||
S: GTATTGTGGGATTAGGA | ||||
D | F: [Btn] GTGTATTGAAGTGGGAGAGAATA | 241 | 19–22 | |
R: CAATAACAATTTTACAAACACAAATAACTT | ||||
S: ATAACTTTTCATTCA | ||||
promoter | A [31] | F: GGATATGTTGGGATAGTT | 98 | 72–83 |
98 CpGs | R: [Btn] CCCAAACACTCACCAAAT | |||
S: GGATATGTTGGGATAGTT |
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Zappe, K.; Pühringer, K.; Pflug, S.; Berger, D.; Böhm, A.; Spiegl-Kreinecker, S.; Cichna-Markl, M. Association between MGMT Enhancer Methylation and MGMT Promoter Methylation, MGMT Protein Expression, and Overall Survival in Glioblastoma. Cells 2023, 12, 1639. https://doi.org/10.3390/cells12121639
Zappe K, Pühringer K, Pflug S, Berger D, Böhm A, Spiegl-Kreinecker S, Cichna-Markl M. Association between MGMT Enhancer Methylation and MGMT Promoter Methylation, MGMT Protein Expression, and Overall Survival in Glioblastoma. Cells. 2023; 12(12):1639. https://doi.org/10.3390/cells12121639
Chicago/Turabian StyleZappe, Katja, Katharina Pühringer, Simon Pflug, Daniel Berger, Andreas Böhm, Sabine Spiegl-Kreinecker, and Margit Cichna-Markl. 2023. "Association between MGMT Enhancer Methylation and MGMT Promoter Methylation, MGMT Protein Expression, and Overall Survival in Glioblastoma" Cells 12, no. 12: 1639. https://doi.org/10.3390/cells12121639