Annals of Surgical Oncology

, Volume 25, Issue 12, pp 3764–3770 | Cite as

Tumor Cell Content and RNA Integrity of Surgical Tissues from Different Types of Tumors and Its Correlation with Ex Vivo and In Vivo Ischemia

  • Xiao-Hui Zheng
  • Shao-Dan Zhang
  • Pei-Fen Zhang
  • Xi-Zhao Li
  • Ye-Zhu Hu
  • Tian Tian
  • Lin Zhu
  • Ruo-Zheng WangEmail author
  • Wei-Hua JiaEmail author
Translational Research and Biomarkers



Tissues from tumor patients are important resources for promoting cancer research, and therefore many biobanks have been established to collect tumor tissues; however, the quality of tumor tissues after surgical resection has not been well documented.


A total of 896 cases of tissues from 12 types of tumors were chosen for this study. First, histopathological examination was conducted to evaluate the tumor cell content; second, microchip electrophoresis was used to determine the RNA integrity number (RIN) in 466 cases of tissues with a tumor cell content ≥ 75%; and, finally, a correlation test was used to analyze the effect of ischemia on RNA integrity in 384 cases of tissues with a recorded ischemia time.


Tumor tissues from 12 different organs had different tumor cell contents and RNA integrity. The liver had the highest percentage (69.7%) of tissue samples with a tumor cell content ≥ 75%, and the highest percentage (96%) of samples with an RIN ≥ 7. RNA integrity was not correlated with limited ex vivo ischemia time (5–60 min) in any of the 12 types of tumors. In contrast, a significant correlation with in vivo ischemia time was observed in several types of tumors.


Not every sample of excised tumor tissue has a sufficient amount of tumor cells and enough RNA integrity. In vivo ischemia has a more significant influence on RNA integrity, and tumor tissues have different tolerances to pre-analytical variables. Those conducting translational research should pay attention to pre-analytical variables when collecting and utilizing tumor tissues.



This work was supported by the National Science Fund for Distinguished Young Scholars (81325018), the Key Project for International Cooperation and Exchange of the National Natural Science Foundation of China (81220108022), the National Key Research and Development Program (2016YFC1302704), and Guangdong Special Support Programs for high-level personnel (2014TX01R201).


No potential conflicts of interest were disclosed.

Supplementary material

10434_2018_6697_MOESM1_ESM.doc (132 kb)
Supplementary material 1 (DOC 131 kb)


  1. 1.
    Garman KS, Nevins JR, Potti A. Genomic strategies for personalized cancer therapy. Hum Mol Genet 2007;16 Spec No. 2:R226–232.CrossRefGoogle Scholar
  2. 2.
    Watson RW, Kay EW, Smith D. Integrating biobanks: addressing the practical and ethical issues to deliver a valuable tool for cancer research. Nat Rev Cancer. 2010;10:646–51.CrossRefGoogle Scholar
  3. 3.
    Vaught J, Rogers J, Myers K, et al. An NCI perspective on creating sustainable biospecimen resources. J Natl Cancer Inst Monogr. 2011;2011:1–7.CrossRefGoogle Scholar
  4. 4.
    Whyte B. National tumor bank set up in United Kingdom. J Natl Cancer Inst. 2003;95(10):706.CrossRefGoogle Scholar
  5. 5.
    LiVolsi VA, Clausen KP, Grizzle W, Newton W, Pretlow TG 2nd, Aamodt R. The cooperative human tissue network. An update. Cancer. 1993;71:1391–94.PubMedGoogle Scholar
  6. 6.
    Myles R, Massett HA, Comey G, Atkinson N, Allsop D, Compton C. Stakeholder research on biospecimen needs and reactions to the development of a national cancer human biobank by the National Cancer Institute. J Natl Cancer Inst Monogr. 2011;2011:16–23.CrossRefGoogle Scholar
  7. 7.
    Le Bastard N, De Deyn PP, Engelborghs S. Importance and impact of preanalytical variables on Alzheimer disease biomarker concentrations in cerebrospinal fluid. Clin Chem. 2015;61:734–43.CrossRefGoogle Scholar
  8. 8.
    Ellervik C, Vaught J. Preanalytical variables affecting the integrity of human biospecimens in biobanking. Clin Chem. 2015;61:914–34.CrossRefGoogle Scholar
  9. 9.
    Moore HM. The NCI biospecimen research network. Biotech Histochem. 2012;87:18–23.CrossRefGoogle Scholar
  10. 10.
    Huang J, Qi R, Quackenbush J, Dauway E, Lazaridis E, Yeatman T. Effects of ischemia on gene expression. J Surg Res. 2001;99:222–27.CrossRefGoogle Scholar
  11. 11.
    Spruessel A, Steimann G, Jung M, et al. Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision. Biotechniques. 2004;36:1030–37.CrossRefGoogle Scholar
  12. 12.
    Imbeaud S, Graudens E, Boulanger V, et al. Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces. Nucl Acids Res. 2005;33:e56.CrossRefGoogle Scholar
  13. 13.
    Morente MM, Mager R, Alonso S, et al. TuBaFrost 2: standardising tissue collection and quality control procedures for a European virtual frozen tissue bank network. Eur J Cancer. 2006;42:2684–91.CrossRefGoogle Scholar
  14. 14.
    Schroeder A, Mueller O, Stocker S, et al. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 2006;7:3.CrossRefGoogle Scholar
  15. 15.
    Mager SR, Oomen MH, Morente MM, et al. Standard operating procedure for the collection of fresh frozen tissue samples. Eur J Cancer. 2007;43:828–34.CrossRefGoogle Scholar
  16. 16.
    Sun H, Sun R, Hao M, Wang Y, Zhang X, Liu Y, et al. Effect of duration of ex vivo ischemia time and storage period on RNA quality in biobanked human renal cell carcinoma tissue. Ann Surg Oncol. 2016;23:297–304.CrossRefGoogle Scholar
  17. 17.
    Lee SM, Schelcher C, Thasler R, Schiergens TS, Thasler WE. Pre-analytical determination of the effect of extended warm or cold ischaemia on RNA stability in the human ileum mucosa. PLoS ONE. 2015;10:e0138214.CrossRefGoogle Scholar
  18. 18.
    Liu NW, Sanford T, Srinivasan R, et al. Impact of ischemia and procurement conditions on gene expression in renal cell carcinoma. Clin Cancer Res. 2013;19:42–9.CrossRefGoogle Scholar
  19. 19.
    Lee SM, Schelcher C, Gashi S, Schreiber S, Thasler RM, Jauch KW, et al. RNA stability in human liver: comparison of different processing times, temperatures and methods. Mol Biotechnol. 2013;53:1–8.CrossRefGoogle Scholar
  20. 20.
    Bao WG, Zhang X, Zhang JG, et al. Biobanking of fresh-frozen human colon tissues: impact of tissue ex vivo ischemia times and storage periods on RNA quality. Ann Surg Oncol. 2013;20:1737–44.CrossRefGoogle Scholar
  21. 21.
    Musella V, Verderio P, Reid JF, et al. Effects of warm ischemic time on gene expression profiling in colorectal cancer tissues and normal mucosa. PLoS ONE. 2013;8:e53406.CrossRefGoogle Scholar
  22. 22.
    Hatzis C, Sun H, Yao H, et al. Effects of tissue handling on RNA integrity and microarray measurements from resected breast cancers. J Natl Cancer Inst. 2011;103:1871–83.CrossRefGoogle Scholar
  23. 23.
    Rudloff U, Bhanot U, Gerald W, et al. Biobanking of human pancreas cancer tissue: impact of ex vivo procurement times on RNA quality. Ann Surg Oncol. 2010;17:2229–36.CrossRefGoogle Scholar
  24. 24.
    Hong SH, Baek HA, Jang KY, et al. Effects of delay in the snap freezing of colorectal cancer tissues on the quality of DNA and RNA. J Korean Soc Coloproctol. 2010;26:316–23.CrossRefGoogle Scholar
  25. 25.
    Bray SE, Paulin FE, Fong SC, et al. Gene expression in colorectal neoplasia: modifications induced by tissue ischaemic time and tissue handling protocol. Histopathology. 2010;56:240–50.CrossRefGoogle Scholar
  26. 26.
    De Cecco L, Musella V, Veneroni S, et al. Impact of biospecimens handling on biomarker research in breast cancer. BMC Cancer. 2009;9:409.CrossRefGoogle Scholar
  27. 27.
    Micke P, Ohshima M, Tahmasebpoor S, Ren ZP, Ostman A, Ponten F, et al. Biobanking of fresh frozen tissue: RNA is stable in nonfixed surgical specimens. Lab Invest. 2006;86:202–11.CrossRefGoogle Scholar
  28. 28.
    Guerrera F, Tabbo F, Bessone L, et al. The influence of tissue ischemia time on RNA integrity and patient-derived xenografts (PDX) engraftment rate in a non-small cell lung cancer (NSCLC) biobank. PLoS ONE. 2016;11:e0145100.CrossRefGoogle Scholar
  29. 29.
    Galissier T, Schneider C, Nasri S, et al. Biobanking of fresh-frozen human adenocarcinomatous and normal colon tissues: which parameters influence RNA quality? PLoS ONE. 2016;11:e0154326.CrossRefGoogle Scholar
  30. 30.
    Sandusky GE, Teheny KH, Esterman M, Hanson J, Williams SD. Quality control of human tissues: experience from the Indiana University Cancer Center-Lilly Research Labs human tissue bank. Cell Tissue Bank. 2007;8:287–95.CrossRefGoogle Scholar
  31. 31.
    Li H, Guo Y, Sun B, Chen K. Histological assessment of tumor tissue samples via the mirror image method. Biopreserv Biobank. 2015;13:25–30.CrossRefGoogle Scholar
  32. 32.
    Asterand C. RNA quality assurance using RIN. Detroit: Asterand; 2006.
  33. 33.
    Ibberson D, Benes V, Muckenthaler MU, Castoldi M. RNA degradation compromises the reliability of microRNA expression profiling. BMC Biotechnol. 2009;9:102.CrossRefGoogle Scholar
  34. 34.
    Lee J, Hever A, Willhite D, Zlotnik A, Hevezi P. Effects of RNA degradation on gene expression analysis of human postmortem tissues. FASEB J. 2005;19:1356–58.CrossRefGoogle Scholar
  35. 35.
    Viana CR, Neto CS, Kerr LM, et al. The interference of cold ischemia time in the quality of total RNA from frozen tumor samples. Cell Tissue Bank. 2013;14:167–73.CrossRefGoogle Scholar
  36. 36.
    Zeugner S, Mayr T, Zietz C, Aust DE, Baretton GB. RNA quality in fresh-frozen gastrointestinal tumor specimens-experiences from the tumor and healthy tissue bank TU Dresden. Recent Results Cancer Res. 2015;199:85–93.CrossRefGoogle Scholar
  37. 37.
    Ma Y, Dai H, Kong X. Impact of warm ischemia on gene expression analysis in surgically removed biosamples. Anal Biochem. 2012;423:229–35.CrossRefGoogle Scholar
  38. 38.
    Fajardy I, Moitrot E, Vambergue A, Vandersippe-Millot M, Deruelle P, Rousseaux J. Time course analysis of RNA stability in human placenta. BMC Mol Biol. 2009;10:21.CrossRefGoogle Scholar
  39. 39.
    Abrahamsen HN, Steiniche T, Nexo E, Hamilton-Dutoit SJ, Sorensen BS. Towards quantitative mRNA analysis in paraffin-embedded tissues using real-time reverse transcriptase-polymerase chain reaction: a methodological study on lymph nodes from melanoma patients. J Mol Diagn. 2003;5:34–41.CrossRefGoogle Scholar
  40. 40.
    Betsou F, Lehmann S, Ashton G, et al. Standard preanalytical coding for biospecimens: defining the sample PREanalytical code. Cancer Epidemiol Biomarkers Prev. 2010;19:1004–11.CrossRefGoogle Scholar
  41. 41.
    Moore HM, Kelly AB, Jewell SD, et al. Biospecimen reporting for improved study quality (BRISQ). Cancer Cytopathol. 2011;119:92–101.CrossRefGoogle Scholar

Copyright information

© Society of Surgical Oncology 2018

Authors and Affiliations

  • Xiao-Hui Zheng
    • 1
    • 2
  • Shao-Dan Zhang
    • 1
  • Pei-Fen Zhang
    • 1
  • Xi-Zhao Li
    • 1
  • Ye-Zhu Hu
    • 1
  • Tian Tian
    • 1
  • Lin Zhu
    • 2
  • Ruo-Zheng Wang
    • 2
    Email author
  • Wei-Hua Jia
    • 1
    • 2
    Email author
  1. 1.Tumor Biobank, Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer MedicineGuangzhouPeople’s Republic of China
  2. 2.Affiliated Tumor Hospital of Xinjiang Medical UniversityÜrümqiPeople’s Republic of China

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