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The Research Of Promoter Methylation In Glioma MGMT Gene

Posted on:2010-10-28Degree:MasterType:Thesis
Country:ChinaCandidate:L B ZhangFull Text:PDF
GTID:2144360278470017Subject:Surgery
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Objective: To explore the relationship among MGMT (O~6-methylguanine-DNA-methyltransferase) promoter methylation in human glioma and MGMT gene expression and chemotherapeutics sensitivities.Methods: The humen glioma cell line u251 was used in this study. The cells were divided into 5-aza-2' -deoxycytidine(5azadC) treated and untreated group. The mRNA level of MGMT gene using RT-PCR was measured in the two groups. The level of MGMT promoter methylation was evaluated with MSP in the 5azadC untreated group. The two groups were treated with cisplatin or temozolomide and the apoptosis of these cells were evaluated with flow cytometry.Results: The MSP results showed that the MGMT gene promoter of glioma cell line u251 had partly methylation. After treated with 5azadC the MGMT gene expression was upregulated. In vitro, sensibility test showed the tolerance to temozolomide while MGMT gene expression increased, however 5azadC together with cisplatin could promote apoptosis of the u251 cells.Conclusion:1. MGMT promoter in human glioma cells existed a phenomenon of partly methylation.2. Promoter methylation might be a main factor of glioma MGMT gene expression downregulation.3. Upregulated MGMT expression might be one of the reasons of temozolomide tolerance.4. 5azadC together with cisplatin could promote apoptosis of the humen glioma cells while MGMT expression was increased.
Keywords/Search Tags:MGMT, methylation, 5-aza-2' -deoxycytidine, MSP, RT-PCR, flow cytometry, cisplatin, temozolomide
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