[A simple and sensitive DNA-RNA hybridization method--in situ hybridization on the nitrocellulose filter for detecting oncogene expression in tumor cells by using digoxigenin-labelled probes].
A simple and sensitive method for detecting oncogene mRNA is performed by fixing tumor cells directly onto nitrocellulose paper, followed by in situ RNA hybridization with digoxigenin-labelled oncogene probes. The human lung large cell carcinoma cell line PLA801-D95 (before and after induction of differentiation with retinoid acid) and leukemic cells from clinical patients were used as target cells. The results showed that 5000 cells were sufficient to give a positive signal for oncogene mRNA. Compared with RNA dot blot, it was found that this technique was quick, simple, sensitive and reproducible. It can be used to study gene expression in tumor cells as well as other cells.
Bao
J
J
Institute of Basic Medical Sciences, Beijing.
D002277:Carcinoma / Q000235:genetics; D004076:Digoxigenin; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007938:Leukemia / Q000235:genetics; D008175:Lung Neoplasms / Q000235:genetics; D009693:Nucleic Acid Hybridization; D015341:Nucleic Acid Probes; D009857:Oncogenes / Q000235:genetics; D014407:Tumor Cells, Cultured
D015341:Nucleic Acid Probes; D004076:Digoxigenin