Impact of ischemia time and storage periods on RNA quality of fresh-frozen breast cancer and esophageal cancer tissue samples in biobank

ZHENG Yang-si, LIN Xuan-hao, LI Fan, XIAO Kun-sheng, CHEN Xi-feng, LIU Chun-peng, YAO Pei-xiu, WANG Shao-hong

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Fudan University Journal of Medical Sciences ›› 2025, Vol. 52 ›› Issue (03) : 437-445. DOI: 10.3969/j.issn.1672-8467.2025.03.015
Method and Technique

Impact of ischemia time and storage periods on RNA quality of fresh-frozen breast cancer and esophageal cancer tissue samples in biobank

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Abstract

Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer （BC） and esophageal cancer （EC） tissue samples in order to establish evidence-based protocols for biobank sample management. Methods The tumor （T） and paired normal （N） tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank， Shantou Central Hospital. Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis. The samples were divided into two groups of <15 min and 15-30 min according to ischemia time， and RNA quality was analyzed at 4 storage periods of 8-10 months （T1）， 14-16 months （T2）， 26-28 months （T3） and 38-40 months （T4）. Results In 96 analyzed samples， 93.8% （90/96） exhibited high quality （RIN≥6）， with 89.6% （43/48） in BC and 97.9% （47/48） in EC. Significant differences in RIN were observed between BC group and EC group （8.050 vs. 8.600， P=0.009）. In EC group， RIN value was significantly negatively correlated with RNA yield （P<0.001）. Moreover， RIN values of tumor-normal pairs exhibited markedly significant differences （7.550 vs. 9.000， P<0.001）. In contrast， no significant difference was detected in BC group （8.200 vs. 7.700， P=0.348）. Statistical analysis showed that RIN value was positively correlated with 28S/18S （P<0.001）， but had no correlation with tumor content （P=0.676） and necrotic content （P=0.055）. Neither ischemia time （<15 min vs. 15-30 min： 8.200 vs. 8.300， P=0.932） nor storage periods （T1-T4： 8.400， 7.700， 8.450， 8.600， P=0.163） compromised RNA quality. Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples. However， limited ischemia time （≤30 min） and long-term storage period （38-40 months） do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

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biobank / RNA quality / ischemia time / storage periods / breast cancer / esophageal cancer

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ZHENG Yang-si , LIN Xuan-hao , LI Fan , et al . Impact of ischemia time and storage periods on RNA quality of fresh-frozen breast cancer and esophageal cancer tissue samples in biobank. Fudan University Journal of Medical Sciences. 2025, 52(03): 437-445 https://doi.org/10.3969/j.issn.1672-8467.2025.03.015

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郑仰思 实验设计和执行，数据整理和分析，论文撰写和修订。林旋豪 实验指导，图表制作。李帆，肖坤胜 样本采集和保存。陈锡丰，刘春鹏 病理制片和诊断。姚佩秀 数据整理和分析。王少洪 论文指导。

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