Detection of keratinocyte growth factor (KGF) transcripts from normal human and archival canine benign prostatic hyperplastic tissues.
This study examined the expression of keratinocyte growth factor (KGF) gene in human and canine prostatic tissues. KGF transcript was detected in normal human prostatic tissues by reverse transcription and polymerase chain reactions (RT-PCR). PCR-generated human KGF complementary DNA (cDNA) clone was confirmed by restriction enzyme digestion analysis and partial DNA sequencing. Expression of KGF in archival canine benign prostatic hyperplastic tissues was also examined. In a pilot experiment, RNAs isolated from formalin-fixed (FF) and formalin-fixed and paraffin-embedded (FFPE) canine prostatic tissues were shown to be of sufficient quality to permit amplification of KGF mRNA by RT-PCR. The transcript of a housekeeping gene, glucose-6-phosphate dehydrogenase (G6PD), was detected by RT-PCR indicating the quality of RNAs to be more than adequate for RNA expression analysis. Later, total RNA from two archival canine FF prostate tissue types, benign prostatic hyperplasis and mild glandular hyperplasia, were used to amplify canine KGF transcripts. Southern hybridization analysis using rat and human KGF cDNAs as probes confirmed the fidelity of the amplified PCR product and it was indeed canine KGF.