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Updated: Jul 19, 2026
Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
Published on: August 1, 2018
Detection of biomarker gene expression by real-time polymerase chain reaction using amplified ribonucleic acids from
Brian K Petroff1, Teresa A Phillips, Bruce F Kimler
1Breast Cancer Prevention Center, University of Kansas Medical Center, Kansas City, Kansas 66160, USA. bpetroff@kumc.edu
Objective:
To address the detection of breast cancer biomarker gene expression in formalin-fixed random periareolar fine needle aspiration (RPFNA) samples of benign breast tissue collected during breast cancer prevention trials by quantitative real-time polymerase chain reaction (qPCR).
Study Design:
Formalin-fixed breast epithelial cells collected by RPFNA and processed as thin layer preparations were isolated by laser capture microdissection (LCM). Ribonucleic acid (RNA) was extracted and amplified using a single round of T7-based linear amplification followed by quality assessment and biomarker assay using TaqMan chemistry.
Results:
More than 80% of RPFNA samples yielded RNA of sufficient quantity and quality for measurement of a panel of biomarker genes following a single round of linear amplification. RNA and protein expression for estrogen receptor alpha, as assessed by LCM/qPCR and immunohistochemistry, were correlated. Amplification plots were similar for cDNA standards and cDNA derived from RPFNA samples.
Conclusion:
Assessment of gene expression using amplified RNA from microdissected formalin-fixed RPFNAs can increase the number of biomarkers used during breast cancer chemoprevention trials.
