PURPOSE: The molecular genetic study of benign prostatic hyperplasia (BPH) requires high quality nucleic acids from BPH tissue. It has been generally assumed, but not experimentally proven, that the diathermy used in trans-urethral resection of the prostate (TURP) damages nucleic acids. MATERIALS AND METHODS: Total RNA, polyA+ RNA and genomic DNA isolated from fresh tissue obtained from TURP procedures were compared with those isolated from tissue obtained from open prostatectomy. RESULTS: On a formaldehyde agarose gel, there was an increase in low molecular weight RNA in the TURP derived samples, in comparison to that in the open prostatectomy derived samples. The 28S and 18S rRNA were present in all TURP specimens and although the two bands were reduced in some samples, there were samples in which the 2 bands were indistinguishable from the open prostatectomy derived samples. Low molecular weight fragments seen in total RNA were reduced greatly in polyA+ RNA and the difference in polyA+ RNA between the TURP and open prostatectomy derived samples was significantly diminished. RT-PCR of two ubiquitously expressed gene transcripts (Galpha11, 256 bp and HPRT, 847 bp) yielded products of similar size and intensity between TURP and open prostatectomy derived samples. Genomic DNA obtained from TURP derived specimens showed no difference in size or enzyme digestibility in comparison to that from open prostatectomy. CONCLUSIONS: Nucleic acids extracted from fresh prostatic tissue derived from TURP procedures can be used as readily as any other fresh tissue for RT-PCR based molecular genetic studies, although polyA+ RNA isolation is recommended if degradation-free RNA is required.
PURPOSE: The molecular genetic study of benign prostatic hyperplasia (BPH) requires high quality nucleic acids from BPH tissue. It has been generally assumed, but not experimentally proven, that the diathermy used in trans-urethral resection of the prostate (TURP) damages nucleic acids. MATERIALS AND METHODS: Total RNA, polyA+ RNA and genomic DNA isolated from fresh tissue obtained from TURP procedures were compared with those isolated from tissue obtained from open prostatectomy. RESULTS: On a formaldehydeagarose gel, there was an increase in low molecular weight RNA in the TURP derived samples, in comparison to that in the open prostatectomy derived samples. The 28S and 18S rRNA were present in all TURP specimens and although the two bands were reduced in some samples, there were samples in which the 2 bands were indistinguishable from the open prostatectomy derived samples. Low molecular weight fragments seen in total RNA were reduced greatly in polyA+ RNA and the difference in polyA+ RNA between the TURP and open prostatectomy derived samples was significantly diminished. RT-PCR of two ubiquitously expressed gene transcripts (Galpha11, 256 bp and HPRT, 847 bp) yielded products of similar size and intensity between TURP and open prostatectomy derived samples. Genomic DNA obtained from TURP derived specimens showed no difference in size or enzyme digestibility in comparison to that from open prostatectomy. CONCLUSIONS: Nucleic acids extracted from fresh prostatic tissue derived from TURP procedures can be used as readily as any other fresh tissue for RT-PCR based molecular genetic studies, although polyA+ RNA isolation is recommended if degradation-free RNA is required.
Authors: Lambertus P W Witte; Christine A Teitsma; Jean J M C H de la Rosette; Martin C Michel Journal: Naunyn Schmiedebergs Arch Pharmacol Date: 2013-11-06 Impact factor: 3.000