Next-gen tissue: preservation of molecular and morphological fidelity in prostate tissue.
BACKGROUND
Personalization of cancer therapy requires molecular evaluation of tumor tissue. Traditional tissue preservation involves formalin fixation, which degrades the quality of nucleic acids. Strategies to bank frozen prostate tissue can interfere with diagnostic studies. PAXgene is an alternative fixative that preserves protein and nucleic acid quality.


METHODS
Portions of prostates obtained from autopsy specimens were fixed in either 10% buffered formalin or PAXgene, and processed and embedded in paraffin. Additional sections were immediately embedded in OCT and frozen. DNA and RNA were extracted from the formalin-fixed, PAXgene-fixed, or frozen tissue. Quantitative PCR was used to compare the quality of DNA and RNA obtained from all three tissue types. In addition, 5 μm sections were cut from specimens devoid of cancer and from prostate cancer specimens obtained at prostatectomy and fixed in PAXgene. They were either stained with hematoxylin and eosin or interrogated with antibodies for p63, PSA and p504.


RESULTS
Comparable tissue morphology was observed in both the formalin and PAXgene-fixed specimens. Similarly, immunohistochemical expression of the P63, PSA and P504 proteins was comparable between formalin and PAXgene fixation techniques. DNA from the PAXgene-fixed tissue was of similar quality to that from frozen tissue. RNA was also amplified with up to 8-fold greater efficiency in the PAXgene fixed tissue compared to the formalin-fixed tissue.


CONCLUSIONS
Prostate specimens fixed with PAXgene have preserved histologic morphology, stain appropriately, and have preserved quality of nucleic acids. PAXgene fixation facilitates the use of prostatectomy tissue for molecular biology techniques such as next-generation sequencing.
Department of Medicine, University of Chicago Chicago, IL.
Marc
M
Gillard
Department of Surgery, University of Chicago Chicago, IL.
Westin R
WR
Tom
Department of Pathology, University of Chicago Chicago, IL.
Tatjana
T
Antic
Department of Pathology, University of Chicago Chicago, IL.
Gladell P
GP
Paner
Department of Pathology, University of Chicago Chicago, IL.
Mark W
MW
Lingen
Department of Medicine, University of Chicago Chicago, IL.
David J
DJ
VanderWeele
PAXgene; Prostate cancer; fixation; formalin; immunohistochemistry; next-generation sequencing