Relevance of the Fanconi anemia pathway in the response of human cells to trabectedin.
Trabectedin (Yondelis; ET-743) is a potent anticancer drug that binds to DNA by forming a covalent bond with a guanine in one strand and one or more hydrogen bonds with the opposite strand. Using a fluorescence-based melting assay, we show that one single trabectedin-DNA adduct increases the thermal stability of the double helix by >20 degrees C. As deduced from the analysis of phosphorylated H2AX and Rad51 foci, we observed that clinically relevant doses of trabectedin induce the formation of DNA double-strand breaks in human cells and activate homologous recombination repair in a manner similar to that evoked by the DNA interstrand cross-linking agent mitomycin C (MMC). Because one important characteristic of this drug is its marked cytotoxicity on cells lacking a functional Fanconi anemia (FA) pathway, we compared the response of different subtypes of FA cells to MMC and trabectedin. Our data clearly show that human cells with mutations in FANCA, FANCC, FANCF, FANCG, or FANCD1 genes are highly sensitive to both MMC and trabectedin. However, in marked contrast to MMC, trabectedin does not induce any significant accumulation of FA cells in G2-M. The critical relevance of FA proteins in the response of human cells to trabectedin reported herein, together with observations showing the role of the FA pathway in cancer suppression, strongly suggest that screening for mutations in FA genes may facilitate the identification of tumors displaying enhanced sensitivity to this novel anticancer drug.
Casado
José A
JA
Division of Hematopoiesis and Gene Therapy Program, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas, Avenida Complutense 22, 28040 Madrid, Spain.
Río
Paula
P
Marco
Esther
E
García-Hernández
Verónica
V
Domingo
Alberto
A
Pérez
Laura
L
Tercero
Juan Carlos
JC
Vaquero
Juan José
JJ
Albella
Beatriz
B
Gago
Federico
F
Bueren
Juan A
JA
D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D002453:Cell Cycle; D004247:DNA / Q000235:genetics / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004149:Dioxoles / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005199:Fanconi Anemia / Q000235:genetics / Q000378:metabolism; D051856:Fanconi Anemia Complementation Group Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology; D044005:Tetrahydroisoquinolines / Q000494:pharmacology*; D013997:Time Factors; D000077606:Trabectedin
D018906:Antineoplastic Agents, Alkylating; D004149:Dioxoles; D051856:Fanconi Anemia Complementation Group Proteins; D044005:Tetrahydroisoquinolines; D016685:Mitomycin; D004247:DNA; D000077606:Trabectedin