Translator Disclaimer
1 February 2007 Using Hoechst 33342 to Target Radioactivity to the Cell Nucleus
Linda S. Yasui, Kai Chen, Ketai Wang, T. Patrick Jones, James Caldwell, Diana Guse, Amin I. Kassis
Author Affiliations +
Abstract

Yasui, L. S., Chen, K., Wang, K., Jones, T. P., Caldwell, J., Guse, D. and Kassis, A. I. Using Hoechst 33342 to Target Radioactivity to the Cell Nucleus. Radiat. Res. 167, 167–175 (2007).

We have explored the use of Hoechst 33342 (H33342) to carry radioactivity to the cell nucleus. H33342 enters cells and targets DNA at adenine-thymine-rich regions of the minor groove. Considerable membrane blebbing and ruffling occur in CHO cells within minutes after its addition to the culture medium in micromolar quantities. Blue vesicles are apparent in the cell cytoplasm, and by 30 min the nuclei are stained dark blue. Upon its binding to DNA, a visible emission shift of the dye can be observed with fluorescence microscopy. We have radioiodinated (125I) H33342 and specifically irradiated nuclear DNA by incubating CHO cells with 125I-H33342 at 37°C and accumulating 125I decays at −90°C. At various times, the cells are thawed and assayed for survival (clonogenicity) and DSB (γ-H2AX) formation. 125I-H33342 decay leads to a monoexponential decrease in cell survival with a D0 of 122 125I decays per cell and a linear increase in DNA DSB induction (equivalent to 15 γ-H2AX foci/cell). Cell death is not modified by the radioprotective effects of H33342 because we use considerably lower concentrations than those that provide a slight protection against γ radiation. We conclude that cell killing by 125I-H33342 and the induction of γ-H2AX foci are highly correlated.

Linda S. Yasui, Kai Chen, Ketai Wang, T. Patrick Jones, James Caldwell, Diana Guse, and Amin I. Kassis "Using Hoechst 33342 to Target Radioactivity to the Cell Nucleus," Radiation Research 167(2), 167-175, (1 February 2007). https://doi.org/10.1667/RR0584.1
Received: 14 February 2006; Accepted: 1 August 2006; Published: 1 February 2007
JOURNAL ARTICLE
9 PAGES


SHARE
ARTICLE IMPACT
RIGHTS & PERMISSIONS
Get copyright permission
Back to Top