In situ hybridization using bromodeoxyuridine labelled DNA probe and RNase H.
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.
Kitazawa
S
S
Kobe University School of Medicine, Second Department of Pathology, Japan.
Kitazawa
R
R
Gotoh
A
A
Fujimori
T
T
Maeda
S
S
D000328:Adult; D001973:Bromodeoxyuridine; D015342:DNA Probes; D015972:Gene Expression Regulation, Neoplastic; D016259:Genes, myc; D006689:Hodgkin Disease / Q000473:pathology; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008297:Male; D010957:Plasmids; D000090063:Proto-Oncogene Mas; D016914:Ribonuclease H
D015342:DNA Probes; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D016914:Ribonuclease H; D001973:Bromodeoxyuridine