Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
复制标题
福尔马林固定石蜡包埋组织保存时间对可提取DNA和RNA数量的影响
DOI：
10.1177/0300060520931259
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发表时间：
2020-06-01
影响因子：
1.6
通讯作者：
Chang，Qing
中科院分区：
文献类型：
--
作者：
Yi，Qing-qing;Yang，Rong;Chang，Qing
目的探讨影响人福尔马林固定石蜡包埋组织(FFPE)不同保存时间DNA和RNA提取量的因素。方法随机抽取2010年、2015年、2017年上海医科大学附属嘉定区中心医院病理科子宫肌瘤合并子宫肌瘤患者的FFPE标本20例。使用DNA/RNA FFPE试剂盒进行DNA和RNA提取。用NanoDrop 2000分光光度计测DNA和RNA浓度及其OD260/OD280比值。用LightCycler480实时定量聚合酶链式反应系统从核酸中扩增出人β珠蛋白基因和乙醛脱氢酶2基因，扩增产物经1%琼脂糖凝胶电泳。结果保存时间越长的标本，核酸提取后DNA和RNA的降解程度越高，DNA和RNA浓度越低。但DNA和RNA纯度无显著差异。99%以上的标本可扩增出β-珠蛋白和ALDH2基因。结论保存时间越长，FFPE组织中可提取DNA和RNA的量越少。然而，这些组织可以用来分析一些小的靶基因。
Objectives This study aimed to investigate the factors affecting the quantity of DNA and RNA extractable from human formalin-fixed paraffin-embedded (FFPE) tissues stored for different lengths of time. Methods We randomly selected 20 FFPE specimens harvested from hysteromyoma patients with uterine fibroids during 2010, 2015, and 2017 at the Department of Pathology, Jiading District Central Hospital Affiliated Shanghai University of Medicine and Health Sciences. DNA and RNA extractions were performed using a DNA/RNA FFPE kit. DNA and RNA concentrations and their OD260/OD280 ratios were determined by a NanoDrop 2000 spectrophotometer. The human β-globin gene and aldehyde dehydrogenase-2 (ALDH2) gene were amplified from nucleic acids using a LightCycler 480 Real-Time PCR System, and PCR amplification products were electrophoresed on 1% agarose gels. Results Specimens that were stored for longer showed more degradation and a reduced concentration of DNA and RNA after nucleic acid extraction. However, there was no significant difference in DNA or RNA purity. β-globin and ALDH2 genes could be amplified from more than 99% of specimens. Conclusion We found that FFPE tissues stored for longer had a reduced quantity of extractable DNA and RNA. However, these tissues could be used for the analysis of some small target genes.