北京天坛医院神经外科人脑肿瘤生物样本库中组织样本RNA的质量评价  

RNA quality evaluation of tissue samples in the human brain tumor biological sample bank of the Department of Neurosurgery of Beijing Tiantan Hospital

作　　者：李奇[1] 乔琳[1] 李子瑞[1] 王雅杰[1] 

机构地区：[1]首都医科大学附属北京天坛医院临床医学研究实验室,100050

出　　处：《中华神经外科杂志》2016年第5期438-442,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（81572474）;北京市中医药科技项目（JJ2015-14）;北京市自然基金（7152098）

摘　　要：目的评价首都医科大学附属北京天坛医院神经外科脑肿瘤生物样本库建库5年来样本RNA的质量。方法收集首都医科大学附属北京天坛医院神经外科2011年1月至2015年12月的脑肿瘤样本，每年度随机选取20份，共100份，使用Trizol试剂提取组织总RNA，检测RNA的纯度、浓度、完整性及样本中GAPDH和ACTB内参基因表达水平。结果2011年度至2015年度样本RNA合格率分别为25％、75％、85％、90％、100％。2011年度样本提取的总RNA大部分纯度和完整性不佳，RNA有部分降解，不能有效扩增内参基因GAPDH和ACTB，Ct值明显高于2012年度至2015年度样本的平均Ct值（P〈0．01）；2012年度至2015年度样本提取的总RNA的A260／A280比值均能达到1．8—2．1，A260／A230比值均能达到2．0以上，大部分样本RNA完整性较好，无降解，80份样本均能进行GAPDH和ACTB基因的有效扩增。结论开展RNA水平的实验研究时，建议使用储存时间在4年以内的样本，且在组织样本使用前进行RNA的质量检测；并应每年定期监测样本质量，在样本出库前进行质量评价。Objective To evaluate the quality of RNA samples since the establishment of the brain tumor bank of the Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University for 5 years. Methods The brain tumor samples were collected from the Department of Neurosurgery, Tiantan Hospital, Capital Medical University between 2011 and 2015. Twenty samples were randomly selected each year （a total of 100 samples）. The total RNA was extracted from tissues by using Trizol reagent, and the RNA purity, concentration, integrity, and GAPDH and ACTB reference gene expression levels in the samples were detected. Results The sample qualification rates of RNA from 2011 to 2015 were 25%, 75%, 85%, 90%, and 100%, respectively. Most of the purity and integrity of the total RNA extracted from the samples in 2011 were poor. RNA had part of the degradation and could not effectively amplify the reference genes GAPDH and ACTB. The Ct value was significantly higher than the mean Ct value of the samples between 2012 and 2015 （ P 〈 0.01 ）. The total ratio of A260/A280 of RNA extracted from the samples between 2012 and 2015 reached 1.8 to 2.1, the ratio of A260/A230 could reach more than 2.0, the integrity of RNA in most of the samples was better, and no degradation was observed. Eighty samples could be used to conduct the effective amplification of GAPDH and ACTB genes. Conclusions When conducting the experiment study of the RNA levels, using the samples of the storage time within 4 years is recommended, the quality detection of RNA should be conducted before using the tissue samples, and the sample quality should be monitored regularly each year. The quality evaluation should be conducted before leaving the sample bank.

关 键 词：脑肿瘤 生物样本库 RNA 质量评价 

分 类 号：R739.41[医药卫生—肿瘤]