Transit tumor retrieval preserves RNA fidelity and obviates snap-freezing.
Genetic expression profiling is enabling investigators to discover new diagnostic and possibly therapeutic pathways in sarcoma biology. To draw substantial conclusions from these molecular analyses, adequate tissue samples must be accrued. Beyond cohort size, the most variable and limiting aspect of doing gene expression analyses on fresh human tissue is the preservation of labile ribonucleic acids extracted from clinical specimens. We have developed a novel retrieval protocol that is readily amenable to the clinical constraints placed on surgeons and pathologists that minimizes variables that can corrupt ribonucleic acid fidelity. We evaluate critically genomic message integrity of mesenchymal tumors derived from transcontinental inter-institutional collaboration. Intact total ribonucleic acid was isolated and assessed for quality and quantity. Ribosomal RNA integrity was quantified using a bioanalyzer. Ribonucleic acid from 42 mesenchymal tumors was isolated and quantified, with selected samples amplified. The mean ribosomal ratios for collaborative institutions ranged from 1.0 to 1.18. Samples remained at 4 degrees C before processing from 1 to 17 days. Tumors stabilized using this protocol retained total ribonucleic acid integrity suitable for amplification and genomic expression analysis regardless of the institutional source or preprocessing duration, enabling a potential consortium of investigators to collaborate in the expression profiling of sarcomas.


METHODS
Diagnostic study, Level III-3 (no consistently applied gold standard). See the Guidelines for Authors for a complete description of levels of evidence.
Randall
R Lor
RL
Hunstman Cancer Institute SARC Lab and Primary Children's Medical Center, Syracuse, NY, USA.
Damron
Timothy A
TA
Coffin
Cheryl M
CM
Bastar
Jeffrey D
JD
Joyner
David E
DE
D001707:Biopsy, Needle; D001859:Bone Neoplasms / Q000235:genetics / Q000473:pathology; D003299:Cooperative Behavior; D015925:Cryopreservation; D019317:Evidence-Based Medicine; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D018212:Giant Cell Tumor of Bone / Q000235:genetics / Q000473:pathology; D006801:Humans; D015588:Observer Variation; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D012334:RNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D012512:Sarcoma, Ewing / Q000235:genetics / Q000473:pathology; D012983:Soft Tissue Neoplasms / Q000235:genetics / Q000473:pathology; D013048:Specimen Handling / Q000379:methods*; D013710:Tendons / Q000473:pathology; D013997:Time Factors
D012333:RNA, Messenger; D012334:RNA, Neoplasm