Ribozyme and free alkylated base: a dual approach for sensitizing Mex+ cells to the alkylating antineoplastic drug.
N-alkyl-nitrosoureas and alkyl-triazenes are alkylating antineoplastic drugs, the efficacy of which is strongly affected by the level of expression of the DNA-repair enzyme O6-methylguanine-DNA methyltransferase (MGMT). In tumors, MGMT activity reduces the chemotherapeutic potential of alkylating drugs; therefore, efforts have been made to down-regulate the protein. A partial sensitization of Mex+ cells to alkylating drugs has been obtained using either free alkylated bases or oligonucleotides targeted against MGMT mRNA. In the present work, O6-methylguanine and a chemically modified ribozyme, without a cationic liposome as a carrier, were coadministered to CHO47 cells, which express a high level of human MGMT protein. The reduction of MGMT mRNA and protein enhanced the genotoxicity of the alkylating drug mitozolomide. Furthermore, the sensitivity of CHO47 cells is the same as that of CHO5 cells, which lack MGMT protein. These data indicate that a strategy in which both mRNA and protein are degradation targets can be successfully applied to down-regulate the MGMT gene.
Mariani
L
L
Istituto di Mutagenesi e Differenziamento, Genetica e Biochimica Tossicologica, Consiglio Nazionale delle Ricerche, Pisa, Italy.
Citti
L
L
Nevischi
S
S
Eckstein
F
F
Rainaldi
G
G
D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D001483:Base Sequence; D016466:CHO Cells / Q000201:enzymology; D003131:Combined Modality Therapy; D006224:Cricetinae; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D015971:Gene Expression Regulation, Enzymologic; D018390:Gene Targeting; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009588:Nitrogen Mustard Compounds / Q000494:pharmacology*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000235:genetics / Q000378:metabolism*; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis / Q000187:drug effects / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012854:Sister Chromatid Exchange / Q000187:drug effects; D014162:Transfection
D018906:Antineoplastic Agents, Alkylating; D017931:DNA Primers; D009588:Nitrogen Mustard Compounds; D016337:RNA, Catalytic; D012333:RNA, Messenger; C040369:mitozolomide; D019853:O(6)-Methylguanine-DNA Methyltransferase