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Comparative Study
. 2006 Apr 25:7:92.
doi: 10.1186/1471-2164-7-92.

Transcriptomes of human prostate cells

Asa J Oudes  1 , Dave S Campbell, Carrie M Sorensen, Laura S Walashek, Lawrence D True, Alvin Y Liu
Affiliations
Comparative Study

Transcriptomes of human prostate cells

Asa J Oudes et al. BMC Genomics. .

Abstract

Background: The gene expression profiles of most human tissues have been studied by determining the transcriptome of whole tissue homogenates. Due to the solid composition of tissues it is difficult to study the transcriptomes of individual cell types that compose a tissue. To overcome the problem of heterogeneity we have developed a method to isolate individual cell types from whole tissue that are a source of RNA suitable for transcriptome profiling.

Results: Using monoclonal antibodies specific for basal (integrin beta4), luminal secretory (dipeptidyl peptidase IV), stromal fibromuscular (integrin alpha 1), and endothelial (PECAM-1) cells, respectively, we separated the cell types of the prostate with magnetic cell sorting (MACS). Gene expression of MACS-sorted cell populations was assessed with Affymetrix GeneChips. Analysis of the data provided insight into gene expression patterns at the level of individual cell populations in the prostate.

Conclusion: In this study, we have determined the transcriptome profile of a solid tissue at the level of individual cell types. Our data will be useful for studying prostate development and cancer progression in the context of single cell populations within the organ.

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Figures

Figure 1
Figure 1
Cell type specific CD antibodies. The specificity of antibodies directed at luminal secretory (CD26), endothelial (CD31), stromal fibromuscular (CD49a), and basal epithelial (CD104) cells are demonstrated. Brown color indicates positive staining. Each prostatic gland is composed of a one-cell layer thick basal epithelium surmounted by columnar secretory cells enclosing the luminal space. The interglandular stroma contains the fibromuscular cells and various other cell types.
Figure 2
Figure 2
Effectiveness of cell sorting. Differential expression of transcripts that code for surface antigens targeted by MACS sorting. (A) luminal (CD26), (B) basal (CD104), (C) stromal (CD49a), and (D) endothelial (CD31). ANOVA results indicate significantly different expression at P < 0.001 (***) and P < 0.01 (**). Cell sorting enriched for the transcripts belonging to the gene that codes for the surface antigen that was the target of cell sorting.
Figure 3
Figure 3
Prostate cell specific genes. Expression levels of genes that are documented in the literature as being expressed by prostate luminal secretory (A), basal epithelial (B), or stromal fibromuscular (C) cells was determined from Affymetrix GeneChip data. ANOVA results indicate significantly different expression at P < 0.001 (***), P < 0.01 (**), and P < 0.05 (*).
Figure 4
Figure 4
p63 Immunohistochemistry. Immunohistochemistry detecting p63 in the prostate. Brown color indicates positive staining. The expression of p63 protein in the prostate is confined to basal cells, which corresponds to the expression of its transcript in our data.
Figure 5
Figure 5
Similarity of prostate cell transcriptomes. A Venn diagram depicting the number of genes expressed by each prostate cell type: basal (red), luminal (green), and stromal (blue). Intermediate colors represent groups of genes that are shared between two cell types. Genes expressed by all prostate cell types are white. The transcriptomes of luminal cells are more similar to basal cells than stromal cells.
Figure 6
Figure 6
Cell type specific expression of TGFβ family members. A heatmap diagram of the expression profiles for genes that belong to the TGFβ and related signaling pathways. Red indicates high expression and blue indicates no detectable expression. Prostate stromal cells differentially express genes involved in TGFβ signaling relative to other prostate cell types.
Figure 7
Figure 7
Transcript expression of BMP signaling pathway members. The diagram indicates cellular locations of differentially expressed genes in the BMP signaling pathway in prostate stromal cells. Decorin, a negative regulator of TGFβ signaling, is produced by stromal cells, which may result in autocrine regulation of the pathway. BMP4 and BMP5 are produced by stromal cells and may be responsible for paracrine regulation of Inhibitor of Differentiation (ID) gene expression in the prostate.
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