DNMT1 mediates chemosensitivity by reducing methylation of miRNA-20a promoter in glioma cells

Experimental & Molecular Medicine
Daoyang ZhouXu-chen Qi

Abstract

Although methyltransferase has been recognized as a major element that governs the epigenetic regulation of the genome during temozolomide (TMZ) chemotherapy in glioblastoma multiforme (GBM) patients, its regulatory effect on glioblastoma chemoresistance has not been well defined. This study investigated whether DNA methyltransferase (DNMT) expression was associated with TMZ sensitivity in glioma cells and elucidated the underlying mechanism. DNMT expression was analyzed by western blotting. miR-20a promoter methylation was evaluated by methylation-specific PCR. Cell viability and apoptosis were assessed using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and TdT-mediated dUTP-biotin nick end labeling assays, respectively. The results showed that compared with parental U251 cells, DNMT1 expression was downregulated, miR-20a promoter methylation was attenuated and miR-20a levels were elevated in TMZ-resistant U251 cells. Methyltransferase inhibition by 5-aza-2'-deoxycytidine treatment reduced TMZ sensitivity in U251 cells. In U251/TM cells, DNMT1 expression was negatively correlated with miR-20a expression and positively correlated with TMZ sensitivity and leucine-rich repeats and immunoglobulin-lik...Continue Reading

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Citations

Jun 12, 2016·Briefings in Functional Genomics·Veronique G LeBlanc, Marco A Marra
Mar 10, 2017·Genes·Duncan Ayers, Jo Vandesompele
Aug 27, 2018·Journal of Cellular Biochemistry·Cigir Biray AvciCumhur Gunduz
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Methods Mentioned

BETA
PCR
PCRs
protein assay
transfection
enzyme-linked immunosorbent assay
Assay
surgical resection
xenograft

Software Mentioned

Motic Images Advanced
Statistical Packages for Social Sciences ( SPSS

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