Analysis of differential gene expression in colorectal cancer and stroma using fluorescence-activated cell sorting purification
| Autor: | Aedín C. Culhane, B. D. Barry, M A Kelly, Henry Paul Redmond, Desmond G. Higgins, Jeffrey Tze-Yee Wang, Thomas G. Cotter, John Calvin Coffey, Myles Smith, Maryanne Donovan, W. O. Kirwan |
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| Rok vydání: | 2009 |
| Předmět: |
Cancer Research
Pathology medicine.medical_specialty Stromal cell Microarray Biopsy colorectal cancer Cell Separation Collagen Type VI Biology Collagen Type I Parenchyma Gene expression medicine Humans RNA Neoplasm gene expression microarray analysis Molecular Diagnostics Oligonucleotide Array Sequence Analysis Regulation of gene expression Tissue Inhibitor of Metalloproteinase-2 Gene Expression Profiling Cell sorting Flow Cytometry Gene expression profiling Gene Expression Regulation Neoplastic Oncology Cancer research Collagen DNA microarray Stromal Cells Colorectal Neoplasms Cell Adhesion Molecules fluorescence-activated cell sorting |
| Zdroj: | British Journal of Cancer |
| ISSN: | 1532-1827 |
| Popis: | Tumour stroma gene expression in biopsy specimens may obscure the expression of tumour parenchyma, hampering the predictive power of microarrays. We aimed to assess the utility of fluorescence-activated cell sorting (FACS) for generating cell populations for gene expression analysis and to compare the gene expression of FACS-purified tumour parenchyma to that of whole tumour biopsies. Single cell suspensions were generated from colorectal tumour biopsies and tumour parenchyma was separated using FACS. Fluorescence-activated cell sorting allowed reliable estimation and purification of cell populations, generating parenchymal purity above 90%. RNA from FACS-purified and corresponding whole tumour biopsies was hybridised to Affymetrix oligonucleotide microarrays. Whole tumour and parenchymal samples demonstrated differential gene expression, with 289 genes significantly overexpressed in the whole tumour, many of which were consistent with stromal gene expression (e.g., COL6A3, COL1A2, POSTN, TIMP2). Genes characteristic of colorectal carcinoma were overexpressed in the FACS-purified cells (e.g., HOX2D and RHOB). We found FACS to be a robust method for generating samples for gene expression analysis, allowing simultaneous assessment of parenchymal and stromal compartments. Gross stromal contamination may affect the interpretation of cancer gene expression microarray experiments, with implications for hypotheses generation and the stability of expression signatures used for predicting clinical outcomes. |
| Databáze: | OpenAIRE |
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