Retroviral insertional mutagenesis as a strategy for the identification of genes associated with cis-diamminedichloroplatinum(II) resistance.
Expression of resistance to cis-diamminedichloroplatinum(II) (CDDP), one of the most effective chemotherapeutic drugs used to treat a variety of malignancies, remains a serious obstacle for improving cancer treatment. To study possible genetic mechanisms underlying the development of CDDP resistance, we have adopted the approach of retroviral insertional mutagenesis. An early-stage CDDP-sensitive human melanoma cell line, WM35, was infected with a defective amphotropic murine retrovirus (murine stem cell virus), and the pooled cells were subsequently selected for CDDP-resistant variants. Nine CDDP-resistant clones independently derived from murine stem cell virus-infected WM35 cells were analyzed and it was found that five of these clones acquired an identical retroviral integration site, designated as CDDP resistance locus 1 (CRL-1), as revealed by isolation of retroviral flanking sequences. Furthermore, using the flanking sequence as probe, we have detected a 3.5-4.0-kilobase message, the expression of which is strongly increased in clones carrying a rearranged CRL-1 locus. These results strongly suggest that overexpression of CRL-1 confers resistance to CDDP in these clones. In addition, the present study indicates that retroviral insertional mutagenesis represents a potential strategy to identify genes responsible for CDDP resistance and possibly other chemotherapeutic drugs as well.
Lu
S J
SJ
Division of Cancer Research, Sunnybrook Health Science Centre, University of Toronto, Ontario, Canada.
Man
S
S
Bani
M R
MR
Adachi
D
D
Hawley
R G
RG
Kerbel
R S
RS
Ben-David
Y
Y
D002945:Cisplatin / Q000494:pharmacology*; D003001:Cloning, Molecular; D004351:Drug Resistance; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics* / Q000821:virology; D016254:Mutagenesis, Insertional; D009693:Nucleic Acid Hybridization; D011533:Proviruses / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics*; D012192:Retroviridae Infections / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D016662:Virus Integration
D002945:Cisplatin