In Situ Hybridization Using Bromodeoxyuridine Labelled DNA Probe and RNase H

Abstract

We developed a modified in situ DNA-RNA hybridization technique and demonstrated c-myc proto-oncogene transcripts in nodular sclerosis type Hodgkin's disease (HD-NS). A hybridization probe was prepared by metabolic labelling of bromodeoxyuridine (BrdU). The hybridized probe was detected with anti-BrdU monoclonal antibody after incubation with ribonuclease H (RNase H), which specifically degrades RNA from DNA-RNA hybrids. In HD-NS, c-myc protooncogene transcripts were demonstrated in the cytoplasm of lacuna cells and a few surrounding lymphoid cells. This modified hybridization method was sensitive and applicable to the study of oncogene expression at a single cell level.

Key Words:

• Bromodeoxyuridine
• in situ hybridization
• RNase H
• c-myc
• Hodgkin's disease