Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Controlled Clinical Trial
. 2012 Jul;176(1):121-32.
doi: 10.1016/j.jss.2011.05.058. Epub 2011 Jun 25.

Differentially expressed genes in window trials are influenced by the wound-healing process: lessons learned from a pilot study with anastrozole

Mary Morrogh  1 , Victor P Andrade, Asawari J Patil, Li-Xuan Qin, Qianxing Mo, Rita Sakr, Crispinita D Arroyo, Edi Brogi, Monica Morrow, Tari A King
Affiliations
Controlled Clinical Trial

Differentially expressed genes in window trials are influenced by the wound-healing process: lessons learned from a pilot study with anastrozole

Mary Morrogh et al. J Surg Res. 2012 Jul.

Abstract

Background: Perioperative window trials provide an opportunity to obtain intact tumor samples at two different time-points for evaluation of potential surrogate biomarkers. We report results of a pilot trial designed to determine if treatment-mediated changes in gene expression can be detected in formalin-fixed paraffin-embedded (FFPE) samples after 10-d exposure to anastrozole in estrogen receptor (ER)-positive breast cancer compared with untreated controls.

Methods: Paired tumor samples (biopsy, surgical) were obtained from 26 postmenopausal women with ER-positive breast cancer. Patients were assigned anastrozole (1 mg/d) for 10 d immediately prior to surgery (13 cases) or no treatment (13 controls). Five hundred two cancer-related genes were examined by the Illumina cDNA-mediated annealing, selection, extension, and ligation, FFPE cDNA array (moderated t-test, P ≤ 0.005). Surrogate biomarkers reflecting changes in gene expression were examined by immunohistochemistry (Wilcoxon rank-based test, P < 0.05).

Results: Sufficient RNA was available from 19 paired samples (8 controls, 11 cases). Frozen tissue and FFPE showed good correlation (r = 0.82). Within each group, 18 genes, reflecting roles in proliferation, angiogenesis, and apoptosis, showed differential expression from biopsy to surgery (P < 0.005). Estrogen-related genes were dysregulated in the treated group only. A reduction in Ki-67 was observed in 7 (54%) treated cases and in 1 (7.7%) control patient.

Conclusions: 10-d exposure to anastrozole resulted in dysregulation of 18/502 cancer-related genes, and Ki-67 was reduced in 54% of cases. FFPE samples demonstrated good correlation with frozen samples. However, changes in gene expression and increased Ki-67 in the control group suggest local effects of wound healing may represent a confounding factor in the interpretation of perioperative window trials.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Scatter plot graphs showing gene expression correlation between fresh-frozen (X-axis) and formalin fixed paraffin-embedded (Y-axis) specimens in patient matched samples #13-control (a), #17-treated (b) and #19-treated (c) representative of the lowest, median, and highest correlation coefficients.
Figure 1
Figure 1
Scatter plot graphs showing gene expression correlation between fresh-frozen (X-axis) and formalin fixed paraffin-embedded (Y-axis) specimens in patient matched samples #13-control (a), #17-treated (b) and #19-treated (c) representative of the lowest, median, and highest correlation coefficients.
Figure 1
Figure 1
Scatter plot graphs showing gene expression correlation between fresh-frozen (X-axis) and formalin fixed paraffin-embedded (Y-axis) specimens in patient matched samples #13-control (a), #17-treated (b) and #19-treated (c) representative of the lowest, median, and highest correlation coefficients.
Figure 2
Figure 2
Box-plot of the 5 genes with significant different variations over time (biopsy to surgery) between the control and treated groups. Boxes represent inferior quartile, median, and superior quartile values for each group.
Figure 2
Figure 2
Box-plot of the 5 genes with significant different variations over time (biopsy to surgery) between the control and treated groups. Boxes represent inferior quartile, median, and superior quartile values for each group.
Figure 2
Figure 2
Box-plot of the 5 genes with significant different variations over time (biopsy to surgery) between the control and treated groups. Boxes represent inferior quartile, median, and superior quartile values for each group.
Figure 2
Figure 2
Box-plot of the 5 genes with significant different variations over time (biopsy to surgery) between the control and treated groups. Boxes represent inferior quartile, median, and superior quartile values for each group.
Figure 2
Figure 2
Box-plot of the 5 genes with significant different variations over time (biopsy to surgery) between the control and treated groups. Boxes represent inferior quartile, median, and superior quartile values for each group.
Figure 3
Figure 3
TUNEL, Ki-67 and CD34 expression changes from biopsy to surgery for individual (a) control and (b) treated patients.
Figure 3
Figure 3
TUNEL, Ki-67 and CD34 expression changes from biopsy to surgery for individual (a) control and (b) treated patients.
Figure 4
Figure 4
Comparison of Ki-67 expression between biopsy and surgery for the control and treated groups. Boxes represent inferior quartile, median, and superior quartile values for each group.
See this image and copyright information in PMC

References

    1. Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet. 2005;365:1687–1717. - PubMed
    1. Chang JC, Makris A, Gutierrez MC, Hilsenbeck SG, Hackett JR, Jeong J, Liu ML, Baker J, Clark-Langone K, Baehner FL, Sexton K, Mohsin S, Gray T, Alvarez L, Chamness GC, Osborne CK, Shak S. Gene expression patterns in formalin-fixed, paraffin-embedded core biopsies predict docetaxel chemosensitivity in breast cancer patients. Breast Cancer Res Treat. 2008;108:233–240. - PubMed
    1. Paik S, Shak S, Tang G, Kim C, Baker J, Cronin M, Baehner FL, Walker MG, Watson D, Park T, Hiller W, Fisher ER, Wickerham DL, Bryant J, Wolmark N. A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer. N Engl J Med. 2004;351:2817–2826. - PubMed
    1. Dixon M. A study looking at the changes in the breast cancer of postmenopausal women having hormone therapy (Primary Endocrine Response Study) Edinburgh: 2011.
    1. Smith I. A trial of a short course of hormone therapy before and after surgery for early breast cancer (POETIC) Multiple cities, UK: 2011.

Publication types

MeSH terms