Expression of human  O  6 ‐methylguanine‐DNA methyltransferase in chinese hamster ovary cells and restoration of cellular resistance to certain  N ‐nitroso compounds | Zendy

Wu Zining | Zendy; Chan ChungLeung | Zendy; Eastman Alan | Zendy; Bresnick Edward | Zendy

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Expression of human O 6 ‐methylguanine‐DNA methyltransferase in chinese hamster ovary cells and restoration of cellular resistance to certain N ‐nitroso compounds

Author(s) -

Wu Zining,

Chan ChungLeung,

Eastman Alan,

Bresnick Edward

Publication year - 1991

Publication title -

molecular carcinogenesis

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.254

H-Index - 97

eISSN - 1098-2744

pISSN - 0899-1987

DOI - 10.1002/mc.2940040612

Subject(s) - chinese hamster ovary cell , biology , transfection , microbiology and biotechnology , o 6 methylguanine dna methyltransferase , plasmid , methyltransferase , carcinogenesis , cell culture , complementary dna , dna , gene , biochemistry , genetics , methylation

We have constructed a plasmid in which the expression of human O 6 ‐methylguanine‐DNA methyltransferase (MGMT) cDNA is driven by the Rous sarcoma virus promoter sequence. Transfection of this plasmid into Chinese hamster ovary (CHO) cells results in expression of MGMT and in cellular resistance to N ‐methyl‐ N ′‐nitro‐ N ‐nitrosoguanidine (MNNG) and 1‐(2‐chloroethyl)‐1‐nitrosourea (CNU), but not to N ‐nitroso‐ N ‐ethylurea. The specific activity of MGMT in transfected CHO cells correlated well with their resistance to MNNG and CNU. Southern analysis showed that the plasmid had been integrated into the CHO cell genome. Western analysis of extracts from transfected CHO cells using an antibody against a peptide corresponding to the carboxyl‐terminal end of the human MGMT protein demonstrated a single band with a molecular size of 24–25 kDa; no such band was observed in extracts from wild‐type CHO cells. These transfected cells may therefore be used to study the role of MGMT in the repair of alkylating DNA lesions and to determine its importance in carcinogenesis as well as in chemotherapy.

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