NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

Stability and Heterogeneity of Expression Profiles in Lung Cancer Specimens Harvested Following Surgical Resection

Author(s): Blackhall Fiona H, Pintilie Melania, Wigle Dennis A, Jurisica Igor, Liu Ni, Radulovich Nikolina, Johnston Michael R, Keshavjee Shaf, Tsao Ming-Sound

Publication: Neoplasia, 2004, Vol. 6, Page 761

PubMed ID: 15720802 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to assess changes in global gene expression and stress and hypoxia-activated genes as a function of (1) delayed sample processing at room temperature, and (2) intratumoral sampling.

Conclusion of Paper

No clear relationship was observed between gene expression and delayed sample processing following surgical resection. Gene expression was highly variable among intratumoral samples for each tumor examined. The authors recommend pooling samples from a minimum of four sites for a single tumor. Samples of tumors that were snap-frozen at increasing time intervals following surgical resection had similar gene expression patterns. Based on these results, the authors conclude that samples snap-frozen within 30 to 60 minutes of surgical resection are suitable for gene expression studies. However, molecular heterogeneity within tumors should be considered as a confounding factor, and sampling and pooling from multiple sites within each tumor may be necessary for expression profiling studies.

Studies

  1. Study Purpose

    The purpose of this study was to determine if gene expression is altered after storage at room temperature (0, 5, 10, 15, 20, 30 , 40 50, 60, 120 min of cold ischemia) following surgical resection prior to snap freezing. Tumor specimens from two patients were used for analysis.

    Summary of Findings:

    Of the 623 genes investigated via microarray there was no clear linear relationship between altered gene expression and delayed processing time. Real-time qRT-PCR analysis of six stress and hypoxia-induced genes (mitogen-activated protein/MAPK10, JNK3; JUN-B proto oncogene, JUN-B; c-Jun proto oncogene, AP-1; carbonic anhydrase IX, CAIX; protease, serine 25, PRSS25; human homologue of Rad6B, HHR6B) confirmed results of the microarray, suggesting that a delay at room temperature of up to 2 h prior to freezing does not alter gene expression.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    RNA DNA microarray
    RNA Real-time qRT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Cold ischemia time 5 min
    10 min
    15 min
    20 min
    30 min
    40 min
    50 min
    60 min
    120 min
    Real-time qRT-PCR Specific Targeted nucleic acid JNK3
    JUN-B
    AP-1
    CAIX
    PRSS25
    HHR6B
  2. Study Purpose

    The purpose of this study was to assess the effect of tumor heterogenity on global and stress and hypoxia-induced gene expression. A total of 6 samples were collected from each of 4 tumors.

    Summary of Findings:

    Gene expression (JNK3, JUNB, AP-1, CAIX, HHR6B, PRSS25) varied widely among the 6 intratumoral samples collected from each of the 4 tumors examined. Variance for all genes except HHR6B was greater among intratumoral samples than those samples subjected to 5-120 min of cold ischemia. Interestingly, gene expression was not correlated to the position of sampling (tumor periphery versus core).

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Cold ischemia time 50 min
    60 min
    120 min
    5 min
    10 min
    15 min
    20 min
    30 min
    40 min
    Biospecimen Aliquots and Components Biospecimen heterogeneity Biospecimen core
    Biospecimen periphery
    RT-PCR Specific Targeted nucleic acid JNK3
    JUNB
    AP-1
    CAIX
    HHR6B
    PRSS25
  3. Study Purpose

    The purpose of this study was to assess the histopathologic appearance of samples collected from 6 different intratumoral locations ranging from the core of the specimen to the periphery. Tumors from 4 patients were examined, and specimens were snap frozen within 30 minutes of resection.

    Summary of Findings:

    Individual tumors displayed consistent histopathology regardless of intratumoral location (tumor periphery versus core), with the percentage of cancer cells ranging from 60-80% per sample.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Morphology H-and-E microscopy
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Cold ischemia time 40 min
    50 min
    60 min
    120 min
    5 min
    10 min
    15 min
    20 min
    30 min
    Biospecimen Aliquots and Components Biospecimen heterogeneity Biospecimen core
    Biospecimen periphery

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