J Cancer 2019; 10(20):4978-4988. doi:10.7150/jca.29372

Research Paper

Impact of Cold Ischemic Time and Freeze-Thaw Cycles on RNA, DNA and Protein Quality in Colorectal Cancer Tissues Biobanking

Xin-Juan Fan1,2, Yan Huang2, Pei-Huang Wu1, Xin-Ke Yin1, Xi-Hu Yu1,3, Xin-Hui Fu1, Li-Li Feng1, Yun-Long Wang1, Hong-Jun Yi2, Zhi-Ting Chen1, Jun-Xiang Yin4, Da-Lu Zhang4, Wei-Xing Feng1, Shao-Mei Bai1, Taewan Kim5, Gordon B. Mills6, Yi-Ling Lu6, Xiang-Bo Wan1,7✉, Lei Wang1,3✉

1. Guangdong Institute of Gastrointestinal, Guangzhou, Guangdong, China.
2. Department of Pathology, the Sixth Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
3. Department of Gastrointestinal Surgery, the Sixth Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
4. China National Center for Biotechnology Development, Beijing, China.
5. The Ohio State University Comprehensive Cancer Center, Columbus, Ohio, USA.
6. Department of Systems Biology, Division of Basic Science Research, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
7. Department of Radiation Oncology, the Sixth Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
XJ Fan, Y Huang, PH Wu and XK Yin contributed equally to this study.

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.

Keywords: colorectal tissue bank, quality control, RNA integrity, DNA integrity, protein expression.

This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions.
How to cite this article:
Fan XJ, Huang Y, Wu PH, Yin XK, Yu XH, Fu XH, Feng LL, Wang YL, Yi HJ, Chen ZT, Yin JX, Zhang DL, Feng WX, Bai SM, Kim T, Mills GB, Lu YL, Wan XB, Wang L. Impact of Cold Ischemic Time and Freeze-Thaw Cycles on RNA, DNA and Protein Quality in Colorectal Cancer Tissues Biobanking. J Cancer 2019; 10(20):4978-4988. doi:10.7150/jca.29372. Available from http://www.jcancer.org/v10p4978.htm