Impact of cold ischemic time and freeze-thaw cycles on RNA, DNA and protein quality in colorectal cancer tissues biobanking

Xin Juan Fan, Yan Huang, Pei Huang Wu, Xin Ke Yin, Xi Hu Yu, Xin Hui Fu, Li Li Feng, Yun Long Wang, Hong Jun Yi, Zhi Ting Chen, Jun Xiang Yin, Da Lu Zhang, Wei Xing Feng, Shao Mei Bai, Taewan Kim, Gordon B. Mills, Yi Ling Lu, Xiang Bo Wan, Lei Wang

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Tissue-derived RNA, DNA and protein samples become more and more crucial for molecular detection in clinical research, personalized and targeted cancer therapy. This study evaluated how to biobanking colorectal tissues through examining the influences of cold ischemic time and freeze-thaw cycles on RNA, DNA and protein integrity. Here, 144 pairs of tumor and normal colorectal tissues were used to investigate the impact of cold ischemic times (0-48h) on RNA, DNA and protein integrity at on ice or room temperature conditions. Additionally, 45 pairs of tissues experienced 0-9 freeze-thaw cycles, and then the RNA, DNA and protein quality were analyzed. On ice, RNA, DNA and protein from colorectal tumor and normal tissues were all stable up to 48h after surgery. At room temperature, RNA in colorectal tumor and normal tissues began to degrade at 8h and 24h, respectively. Meanwhile, the tumor tissues DNA degradation occurred at 24h after surgery at room temperature. Similarly, the protein expression level of tumor and normal tissues began to change at 24h after the surgery at room temperature. Interestingly, tissue RNA and DNA remained stable even after 9 freeze-thaw cycles, whereas the proteins levels were remarkably changed after 7 freeze-thaw cycles. This study provided a useful evidence on how to store human colorectal tissues for biobanking. Preserving the surgical colorectal tissue on ice was an effective way to prevent RNA, DNA and protein degradation. Importantly, more than 7 repeated freeze-thaw cycles were not recommended for colorectal tissues.

    Original languageEnglish (US)
    Pages (from-to)4978-4988
    Number of pages11
    JournalJournal of Cancer
    Volume10
    Issue number20
    DOIs
    StatePublished - 2019

    Keywords

    • Colorectal tissue bank
    • DNA integrity
    • Protein expression
    • Quality control
    • RNA integrity

    ASJC Scopus subject areas

    • Oncology

    Fingerprint Dive into the research topics of 'Impact of cold ischemic time and freeze-thaw cycles on RNA, DNA and protein quality in colorectal cancer tissues biobanking'. Together they form a unique fingerprint.

    Cite this