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Updated: Mar 18, 2026
Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts
Published on: April 29, 2014
Targeted Transcriptional Profiling of Microdissected Biopsy Specimens Representing Early Colonic Neoplasia
Allen Mo1, Stephen Jackson2, Thomas J Devers3
1Center for Molecular Medicine, UConn Health, Farmington, Connecticut.
Abstract:
Our incomplete understanding of the critical changes that accompany the earliest stages of tumor initiation provides a substantial hurdle for the development of novel intervention strategies for cancer prevention. Premalignant lesions are inherently difficult to characterize given their diminutive size, creating technical obstacles for accurate genetic profiling. Here, we describe an approach combining laser-capture microdissection (LCM) with targeted RNA-sequencing to study the transcriptional state of epithelial and stromal cells during the earliest detectable stage of human colorectal neoplasia, the aberrant crypt foci (ACF). We provide a robust and reproducible workflow for RNA isolation, library preparation, and expression profiling of laser-captured cells from frozen OCT-embedded tissue specimens. It is anticipated that the methodological approach outlined in this report will provide a framework for a broad range of microgenomics analyses that can be routinely applied to many other premalignant tissues. J. Cell. Biochem. 117: 2677-2681, 2016. © 2016 Wiley Periodicals, Inc.
Insights
Studying early cancer development is hard. This research uses laser-capture microdissection and RNA sequencing to analyze gene activity in the earliest colorectal neoplasia (aberrant crypt foci) for better cancer prevention.
Area of Science:
- Oncology
- Genomics
- Molecular Biology
Background:
- Understanding early tumor initiation is crucial for cancer prevention, but premalignant lesions are difficult to study.
- Accurate genetic profiling of small lesions presents significant technical challenges.
Purpose of the Study:
- To develop and validate a method for analyzing the transcriptional state of cells in early colorectal neoplasia.
- To investigate the earliest detectable stage of human colorectal neoplasia, the aberrant crypt foci (ACF).
Main Methods:
- Combined laser-capture microdissection (LCM) with targeted RNA-sequencing.
- Developed a workflow for RNA isolation, library preparation, and expression profiling of laser-captured cells.
- Utilized frozen OCT-embedded tissue specimens.
Main Results:
- Successfully profiled the transcriptional state of epithelial and stromal cells within ACF.
- Established a robust and reproducible workflow for microgenomic analysis of small tissue samples.
Conclusions:
- The described methodological approach enables detailed analysis of early premalignant lesions.
- This workflow provides a framework for microgenomics analyses applicable to various premalignant tissues.
- Facilitates advancements in understanding and preventing cancer initiation.
