Quality of nucleic acids extracted from fresh prostatic tissue obtained from TURP procedures
Quality of nucleic acids extracted from fresh prostatic tissue obtained from TURP procedures
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DOI：
10.1016/s0022-5347(05)67945-1
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发表时间：
2000-02-01
影响因子：
6.6
通讯作者：
Dong, QH
中科院分区：
文献类型：
--
作者：
Margan, SH;Handelsman, DJ;Dong, QH
目的：良性前列腺增生（BPH）的分子遗传学研究需要来自 BPH 组织的高质量核酸。人们普遍认为，但未经实验证明，经尿道前列腺切除术 (TURP) 中使用的透热疗法会损坏核酸。 材料和方法：将从 TURF 手术获得的新鲜组织中分离的总 RNA、polyA(+) RNA 和基因组 DNA 与从开腹前列腺切除术获得的组织中分离的总 RNA、polyA(+) RNA 和基因组 DNA 进行比较。 结果：在甲醛琼脂糖凝胶上，与 TURF 来源的样品相比，在甲醛琼脂糖凝胶上，TURF 来源的样品中的低分子量 RNA 有所增加。开放性前列腺切除术衍生的样本。 28S 和 18S rRNA 存在于所有 TURF 标本中，尽管这两个条带在某些样品中减少，但在某些样品中，这 2 个条带与开放性前列腺切除术衍生的样品无法区分。总RNA中所见的低分子量片段在polyA(+)RNA中大大减少，并且TURF和开放性前列腺切除术衍生的样品之间的polyA(+)RNA差异显着减小。两种普遍表达的基因转录本（G α(11)，256 bp 和 HPRT，847 bp）的 RT-PCR 在 TURF 和开放性前列腺切除术衍生的样品之间产生了相似大小和强度的产物。与开放性前列腺切除术相比，从 TURF 来源的标本中获得的基因组 DNA 在大小或酶消化率方面没有差异。 结论：从 TURF 程序中提取的新鲜前列腺组织中提取的核酸可以像任何其他新鲜组织一样方便地用于基于 RT-PCR 的分子遗传学研究，但如果需要无降解的 RNA，建议分离多聚 A(+) RNA。
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 TURF 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 TURF derived samples, in comparison to that in the open prostatectomy derived samples. The 28S and 18S rRNA were present in all TURF 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 TURF and open prostatectomy derived samples was significantly diminished. RT-PCR of two ubiquitously expressed gene transcripts (G alpha(11), 256 bp and HPRT, 847 bp) yielded products of similar size and intensity between TURF and open prostatectomy derived samples. Genomic DNA obtained from TURF 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 TURF 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.