Staining of Proliferating Cell Nuclei with Biotinylated dUTP

A method of in situ biotinylated dUTP incorporation into nuclei was developed for evaluaton of cell growth activity. HeLa cells or frozen sections of cancer tissues were incubated in a solution containing bio-dUTP and the other deoxyribonucleotides (dATP, dCTP, dGTP), after permeabilization of cell membrane with Triton X-100, and signals from bio-dUTP in nuclei were detected with an avidin-biotin-peroxidase complex (ABC method), or with fluoresceinavidin. This staining procedure was simple and rapid as compared with that for BrdU incorporated into DNA. In HeLa cells, bio-dUTP-incorporated cells could be stained by the ABC method and the labeling index was compatible with that of BrdU, but the intranuclear distribution patterns of bio-dUTP-incorporated cells was also possible. In some frozen sections of cancer tissues, cells synthesizing DNA were labeled with bio-dUTP. These results suggest that this will be a useful method for evaluating cell growth activity, although the biological significance may be different the BrdU incorporation experiment.

Biotinylated dUTP

BrdU

DNA systhesis

Flow cytometry