Retroviral insertional mutagenesis as a strategy for the identification of genes associated with cis-diamminedichloroplatinum(II) resistance.

摘要: Expression of resistance to cis -diamminedichloroplatinum(II) (CDDP), one the most effective chemotherapeutic drugs used treat a variety malignancies, remains serious obstacle for improving cancer treatment. To study possible genetic mechanisms underlying development CDDP resistance, we have adopted approach retroviral insertional mutagenesis. An early-stage CDDP-sensitive human melanoma cell line, WM35, was infected with defective amphotropic murine retrovirus (murine stem virus), and pooled cells were subsequently selected CDDP-resistant variants. Nine clones independently derived from virus-infected WM35 analyzed it found that five these acquired an identical integration site, designated as locus 1 ( CRL-1 ), revealed by isolation flanking sequences. Furthermore, using sequence probe, detected 3.5–4.0-kilobase message, expression which is strongly increased in carrying rearranged locus. These results suggest overexpression confers clones. In addition, present indicates mutagenesis represents potential strategy identify genes responsible possibly other well.