Abstract:
BACKGROUND:O(6)-methylguanine-DNA-methyltransferase (MGMT) is a specific DNA revising enzyme transferring alkylated groups from DNA to its cysteine residue to avoid the abnormal twisting of DNA. Therefore, it is one of the drug resistant genes targeted in the treatment of cancer. This study explored the protective effect of MGMT gene transferred into mammalian cells. METHODS:Mammalian expression vector containing the MGMT gene cloned from human hepatocytes by RT-PCR was constructed and transferred into K562 cells and human peripheral blood mononuclear cells (PBMCs) via liposome, then assayed for gene expression at RNA and protein levels. MTT assay was used to check the drug resistance of cells transfected with MGMT gene. RESULTS:MGMT gene was successfully cloned. Real-time PCR showed that the mRNA expression in gene transfected groups in K562 cell line and PBMC were 13.4 and 4.0 times that of the empty vector transfected groups respectively. RESULTS:of Western blotting showed distinct higher expression of MGMT in gene transfected group than in other two groups. The IC(50) values increased to 7 and 2 times that of the original values respectively in stable transfected K562 cells and transient transfected PBMC. CONCLUSION:The alkylating resistance of eukaryotic cells is enhanced after being transfected with MGMT gene which protein product performs the protective function, and may provide the reference for the protective model of peripheral blood cells in cancer chemotherapy.
journal_name
Chin Med J (Engl)journal_title
Chinese medical journalauthors
Li DB,Wang JS,Fang Q,Sun HY,Xu W,Li WDsubject
Has Abstractpub_date
2007-04-20 00:00:00pages
714-7issue
8eissn
0366-6999issn
2542-5641journal_volume
120pub_type
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journal_title:Chinese medical journal
pub_type: 杂志文章,评审
doi:10.1097/CM9.0000000000000628
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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pub_type: 杂志文章
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journal_title:Chinese medical journal
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:10.4103/0366-6999.187845
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 临床试验,杂志文章,随机对照试验
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:10.1097/CM9.0000000000000619
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
更新日期:2000-06-01 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:10.1097/CM9.0000000000000002
更新日期:2019-01-05 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 临床试验,杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:10.4103/0366-6999.159354
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
更新日期:2012-03-01 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
更新日期:2014-01-01 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章,多中心研究
doi:10.4103/0366-6999.223860
更新日期:2018-02-05 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
更新日期:2012-11-01 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
更新日期:2011-07-05 00:00:00
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:
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