Mizuno T, Nagamura H, Iwamoto K S, Ito T, Fukuhara T, Tokunaga M, Tokuoka S, Mabuchi K, Seyama T
Department of Radiobiology, Radiation Effects Research Foundation, Hiroshima, Japan.
Diagn Mol Pathol. 1998 Aug;7(4):202-8. doi: 10.1097/00019606-199808000-00004.
The validity of molecular studies using DNA and RNA extracted from decades-old formalin-fixed and paraffin-embedded tissue blocks has been demonstrated. The quality and usability of DNA and RNA from archival tissues are modified by various factors, such as the fixative, the fixation time, and the postmortem time. However, in contrast to DNA, there are no comprehensive studies quantitatively addressing the feasibility of RNA from old (more than 10 years) archival samples. This study examined the integrity of RNA extracted from 738 autopsy liver and 63 autopsy thyroid cancer tissue blocks procured during a span of nearly four decades, beginning in 1952 and ending in 1989, from the atomic bomb survivors. The integrity of RNA was assessed by amplification of c-BCR messenger RNA (mRNA) between two sequential exons with an intervening intron by reverse-transcription polymerase chain reaction (RT-PCR). The integrity of RNA was influenced by the age of the samples and the postmortem time, but not by the formalin-fixation period. It was possible to amplify more than 60% of the samples. Using these RNAs, the HCV genome in liver cancers and the H4-RET gene in thyroid cancers were detectable. This study illustrates the possibility of molecular studies using RNA from routinely prepared paraffin blocks stored for long periods and provides the statistics and critical factors to consider in assessing the feasibility of such contemplated studies.
使用从数十年前福尔马林固定石蜡包埋组织块中提取的DNA和RNA进行分子研究的有效性已得到证实。存档组织中DNA和RNA的质量和可用性会受到多种因素的影响,如固定剂、固定时间和死后时间。然而,与DNA不同的是,目前尚无全面的研究定量探讨来自旧的(超过10年)存档样本的RNA的可行性。本研究检测了从1952年开始至1989年近四十年间收集的738份原子弹爆炸幸存者尸检肝脏组织块和63份尸检甲状腺癌组织块中提取的RNA的完整性。通过逆转录聚合酶链反应(RT-PCR)扩增两个连续外显子之间带有间隔内含子的c-BCR信使RNA(mRNA)来评估RNA的完整性。RNA的完整性受样本年龄和死后时间的影响,但不受福尔马林固定时间的影响。超过60%的样本能够进行扩增。利用这些RNA,可以检测到肝癌中的HCV基因组和甲状腺癌中的H4-RET基因。本研究说明了使用从长期保存的常规制备石蜡块中提取的RNA进行分子研究的可能性,并提供了评估此类预期研究可行性时需要考虑的统计学数据和关键因素。