Activation of DNA damage response pathways as a consequence of anthracycline-DNA adduct formation

作者：Forrest Robert A; Swift Lonnie P; Rephaeli Ada; Nudelman Abraham; Kimura Ken Ichi; Phillips Don R; Cutts Suzanne M^*

来源：Biochemical Pharmacology, 2012, 83(12): 1602-1612.

DOI：10.1016/j.bcp.2012.02.026

• Doxorubicin
• Formaldehyde
• DNA damage
• ATR
• ATM

The cytotoxicity of doxorubicin, a clinically used anti-neoplastic drug, can be enhanced by formaldehyde (either endogenous or exogenous) to promote the formation of doxorubicin-DNA adducts. Formaldehyde supplies the carbon required for the covalent linkage of doxorubicin to one strand of DNA, with hydrogen bonds stabilising the doxorubicin mono-adduct to the other strand of DNA, to act much like an interstrand crosslink. Interstrand crosslinks present a major challenge for cellular repair processes, requiring the activation of numerous DNA damage response proteins for resolution of the resulting DNA intermediates and damage. This work investigates DNA damage response proteins activated by doxorubicin-DNA adducts. Although p53 was phosphorylated at Serine 15 in response to adducts, long term growth inhibition of mammalian cells was not affected by p53 status. Using siRNA technology and kinase inhibitors we observed enhanced cellular sensitivity to doxorubicin-DNA adducts when the activity of the signalling protein kinases ATM and ATR were lost. Cells synchronised using a double thymidine block were sensitised to adduct-initiated cell death upon ATR knockdown, but relatively unaffected by ATM knockdown. Loss of ATR was associated with abrogation of a drug-induced G(2)/M block and induction of mitotic catastrophe, while loss of ATM was associated with drug-induced apoptosis in non-synchronised cells. These proteins may therefore be potential drug targets to achieve synergistic cytotoxic responses to doxorubicin-DNA adduct forming therapies. The analysis of these protein kinases with respect to cell cycle progression indicates that AIR is required for G2/M checkpoint responses while ATM appears to function in G(1) mediated responses to anthracycline adducts.

• 出版日期2012-6-15