[Reproducibility of nonradioactive DDRT-PCR method for detection of gene expression in squamous epithelial carcinoma cells of the upper aerodigestive tract].

BACKGROUND: The analysis of the gene spectrum in tumor and normal cells may provide information about genes involved in the differentiation or in the genesis of tumors. Differential Display Reverse Transcriptase Polymerase Chain Reaction (DDRT-PCR) is an innovative method that enables quick analysis of almost all mRNA molecules expressed in the cells.
METHODS: In the trials, the mRNAs of keratinocytes and tumor cells were amplified by a number of oligonucleotide pairs after reverse transcription and resolved on polyacrylamide gel. A modified protocol for the amplification of cDNA probes was devised for detecting PCR products with silver nitrate.
RESULTS: After reverse transcription of mRNAs, all cDNA samples were successfully amplified using the protocol devised for the silver nitrate DDRT-PCR, and the differentially expressed fragments were reproducibly demonstrated.
CONCLUSIONS: The high reproducibility and feasibility of silver nitrate DDRT-PCR expand the alternatives for analyzing gene expression and detecting selectively expressed genes in different kinds of cells.