Abstract 5493: Lethal mutagenesis of cancer cells by nucleoside analogs
2011; American Association for Cancer Research; Volume: 71; Issue: 8_Supplement Linguagem: Inglês
10.1158/1538-7445.am2011-5493
ISSN1538-7445
AutoresEdward Fox, Jiang‐Cheng Shen, Lawrence A. Loeb,
Tópico(s)Cancer-related Molecular Pathways
ResumoAbstract The evolution of cancer and RNA viruses is characterized by mutation accumulation facilitating phenotypic plasticity and environmental adaptation. We have demonstrated that serial passage of virus from HIV-infected human lymphoblastoid cells in the presence of mutagenic nucleosides results in mutation accumulation, leading to error catastrophe, and viral extinction. In limited phase II clinical studies, administration of a cytidine analog, KP-1461, to individuals infected with HIV results in increased viral mutations. We hypothesize that cancer cells treated with mutagenic nucleoside analogs will progressively accumulate mutations until the error threshold for viability is exceeded and the tumor ablated. We have serially passaged mismatch repair proficient and deficient colon cancer cells in the presence of 36 different mutagenic deoxynucleoside analogs and established a new protocol for measuring both mis-incorporation and repair during DNA synthesis in cells. DNA templates, purified from phage, are blocked with terminal non-hydrolysable nucleotides to prevent degradation by cellular nucleases. After transfection of double-stranded DNA into human cells, we can recover >50,000 molecules/cell. After transfection of single-stranded DNA molecules hybridized to a 5′-biotin labeled primer, extended primers are recovered on streptavidin beads; the frequency of misincorporation into the newly synthesized DNA varies from 10-3 to 10-6, depending on the genetic background of the tranfected cells. The versatility of this assay will facilitate studies on the efficiency of DNA repair, the role of different human DNA polymerases in DNA synthesis and bypass of DNA damage, and the stratification of individuals with respect to DNA repair capacity and fidelity. Enhancing the mutation frequency of cancer cells represents a new avenue for treatment, particularly for patients with tumors that have accumulated large numbers of mutations and/or who failed extensive chemotherapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5493. doi:10.1158/1538-7445.AM2011-5493
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