低温冻存胃肠道肿瘤样本对其生物大分子的影响  

Effect of Cryopreservation of Gastrointestinal Tumor Samples on Their Biomacromolecules

作　　者：吴彬 孙溶励 杨心妮 赵明洲 李玉强 Wu Bin;Sun Rongli;Yang Xinni;Zhao Mingzhou;Li Yuqiang(Center for Clinical Biological Samples,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000 China)

机构地区：[1]锦州医科大学附属第一医院临床生物样本中心,辽宁锦州121000

出　　处：《锦州医科大学学报》2022年第4期6-13,共8页Journal of Jinzhou Medical University

基　　金：辽宁省青年科技人才“育苗”项目,项目编号:JYTQN2020022。

摘　　要：目的探讨胃肠道肿瘤组织样本经低温冻存后对其生物大分子DNA、RNA及蛋白质的影响。方法随机选取2014—2021年间锦州医科大学附属第一医院临床生物样本中心采集、冻存的胃癌及结直肠癌样本的癌组织、癌旁组织及正常组织96份,并依据低温冻存年限分为≤2年、3~5年和≥6年3组。采用分光光度计测量DNA、RNA的纯度及蛋白质浓度,1%琼脂糖电泳检测DNA完整性并进行质量分级,测定RNA分子完整数(RIN值)评估RNA完整性,考马斯亮蓝法测定蛋白质完整性并进行质量分级。结果3组低温冻存年限的胃肠道肿瘤组织之间DNA纯度和完整性、RNA纯度以及蛋白质浓度均无显著性差异(P>0.05),胃癌及结直肠癌样本的DNA、RNA及蛋白质完整性无显著性差异(P>0.05)。胃肠道肿瘤样本正常组织的DNA完整性与癌组织及癌旁组织均有显著性差异(P<0.05)。组织样本的RIN值显示,存储≥6年的样本RNA完整性及蛋白质完整性较存储3~5年及≤2年均有显著下降(P<0.05)。结论长期低温冻存的胃肠道肿瘤样本对其DNA的影响较小,冻存超过6年RNA和蛋白质会开始发生降解。Objective To explore the influence of gastrointestinal tumor tissue samples after cryopreservation on their biological macromolecule DNA,RNA and protein.Methods A total of 96 cancerous tissues,para-cancerous tissues and normal tissues of gastric cancer and colorectal cancer collected and cryopreserved at the Center for Clinical Biological Sample of the First Affiliated Hospital of Jinzhou Medical University from 2014 to 2021 were randomly selected.According to the cryopreservation period,they were divided into three groups:≤2 years,3~5 years and≥6 years.DNA and RNA purity and protein concentration were measured by spectrophotometer,DNA integrity was detected by 1%agarose electrophoresis and quality grading was performed,RNA molecular integrity number(RIN)was determined to evaluate RNA integrity,and protein integrity and quality grading were determined by Coomash bright blue method.Results There were no significant differences in DNA purity and integrity,RNA purity and protein concentration among the three groups of gastrointestinal tumor tissues cryopreserved for years(P>0.05).The DNA,RNA and protein integrity of gastric cancer and colorectal cancer samples were no significant differences(P>0.05).The DNA integrity of normal tissues of gastrointestinal tumor samples was significantly different from cancerous tissues and para-cancerous tissues(P<0.05).The RIN value of tissue samples showed that RNA integrity and protein integrity of samples stored for≥6 years were significantly lower than those stored for 3~5 years and≤2 years(P<0.05).Conclusion Long-term cryopreservation of gastrointestinal tumor samples has little effect on their DNA and RNA.Protein will begin to degrade after cryopreservation for more than 6 years.

关 键 词：胃肠道肿瘤 低温冻存 生物大分子 纯度 完整性 

分 类 号：R735[医药卫生—肿瘤]