Sun Hongyan, Sun Ran, Hao Miao, Wang Yuqian, Zhang Xuewen, Liu Ya, Cong Xianling
Tissue Bank, China-Japan Union Hospital of Jilin University, Jilin, China.
Department of Nutrition and Food Hygiene, School of Public Health of Jilin University, Jilin, China.
Ann Surg Oncol. 2016 Jan;23(1):297-304. doi: 10.1245/s10434-014-4327-9. Epub 2015 Jan 8.
RNA degradation is a major problem in tissue banking, and the effects of the ex vivo ischemia time, storage time, and transport conditions on RNA integrity and gene expression have not been well understood.
A total of 100 fresh-frozen clear cell carcinoma tissues and matched normal tissues during five storage periods (≤6, 7-12, 13-18, 19-24, and 25-30 months) were chosen to detect RNA quality. At surgery, fresh kidney cancer tissues from five patients were cut into pieces and snap frozen. Additional fresh tissue pieces were (1) left at room temperature, (2) kept on ice, or (3) placed in normal saline before being snap frozen after 0.5, 1, 2, or 4 h. RNA integrity was determined by microchip electrophoresis, and gene expression was analyzed by real-time polymerase chain reaction.
Altogether, 82 % of kidney cancer specimens banked using a standardized protocol yielded RNA with an RNA integrity number of ≥7 (7.77 ± 0.95, 7.87 ± 0.37, 7.15 ± 1.46, 8.10 ± 0.64, and 7.11 ± 1.08 during the five storage periods, respectively). RNA remained intact after 4 h on ice, whereas degradation was found in tissues left at room temperature or in saline. Expression of genes in certain functional pathways changed during storage under three conditions.
More than 80 % of the banked kidney cancer biospecimens collected following a standardized protocol yielded high-quality RNA. Fresh human cancer tissue samples should be transported on ice before biobanking to avoid a major reduction in RNA quality. The presented data should be considered in attempts to further standardize tissue biospecimen collection and banking.
RNA降解是组织库中的一个主要问题，而体外缺血时间、储存时间和运输条件对RNA完整性及基因表达的影响尚未得到充分了解。
选取100份新鲜冷冻的透明细胞癌组织及配对的正常组织，分五个储存期（≤6个月、7 - 12个月、13 - 18个月、19 - 24个月和25 - 30个月）检测RNA质量。手术时，将5例患者的新鲜肾癌组织切成小块并速冻。另外的新鲜组织块分别进行如下处理：(1) 室温放置；(2) 置于冰上；(3) 放入生理盐水中，分别在0.5、1、2或4小时后速冻。通过微芯片电泳测定RNA完整性，采用实时聚合酶链反应分析基因表达。
总体而言，按照标准化方案保存的肾癌标本中，82%产生的RNA完整性数值≥7（五个储存期的数值分别为7.77±0.95、7.87±0.37、7.15±1.46、8.10±0.64和7.11±1.08）。置于冰上4小时后RNA仍保持完整，而室温放置或置于盐水中的组织出现降解。在三种条件下储存期间，某些功能途径中的基因表达发生了变化。
按照标准化方案收集的肾癌生物标本中，超过80%产生了高质量RNA。新鲜的人类癌组织样本在生物样本库储存前应在冰上运输，以避免RNA质量大幅下降。在进一步规范组织生物样本采集和储存时应考虑本文所呈现的数据。
