Transcriptome analysis to identify the downstream genes of androgen receptor in dermal papilla cells

Abstract

Background: Testosterone signaling mediates various diseases, such as androgenetic alopecia and prostate cancer. Testosterone signaling is mediated by the androgen receptor (AR). In this study, we fortuitously found that primary and immortalized dermal papilla cells suppressed AR expression, although dermal papilla cells express AR in vivo. To analyze the AR signaling pathway, we exogenously introduced the AR gene via a retrovirus into immortalized dermal papilla cells and comprehensively compared their expression profiles with and without AR expression.

Results: Whole-transcriptome profiling revealed that the focal adhesion pathway was mainly affected by the activation of AR signaling. In particular, we found that caveolin-1 gene expression was downregulated in AR-expressing cells, suggesting that caveolin-1 is controlled by AR.

Conclusion: Our whole transcriptome data is critical resources for discovery of new therapeutic targets for testosterone-related diseases.

Keywords: Androgen receptor; Dermal papilla cells; RNA-Seq.

Fig. 1

Workflow of RNA-Seq analysis and PCA of immortalized human dermal papilla cells (HFDPCs) with K4DT cells and AR-expressing cells. A Workflow of the analysis. B Correlation matrix of all samples. Triplicate samples formed unique clusters. C Mapping ratio of parent immortalized HFDPC-K4DT cells and AR-expressing HFDPC-K4DT cells. The mapping results of parent K4DT were reproduced from our previous publication (Fukuda T., et al., 24: 101929, iScience, 2012). D PCA of expression profiling of HFDPC-K4DT cells with and without AR expression

Fig. 2

Heatmap analysis of differentially expressed genes listed in focal adhesion pathway and Proteoglycans in cancer pathway, which are listed in the KEGG database. Red indicates high expression and blue indicates low expression of genes

Fig. 3

Expression level of caveolin-1 and EGF receptors detected by qRT-PCR in identical RNAs, which used for RNA-Seq. Relative RNA quantality which detected with internal control (GAPDH) were shown in the graph. Average value and standard errors are shown with biological triplicated samples

Fig. 4

Expression level of caveolin-1 and EGF receptors in HFDPC-K4DT parental cell, and AR expressing HFDPC-K4DT cell, and after the treatment of 50 nM of DHT treatment. A Expression level of caveolin-1, before treatment of DHT (Left side), and after the 8 h of DHT treatment (Right side). B Expression level of EGFR, before before treatment of DHT (Left side), and after the 8 h of DHT treatment (Right side). Average value and standard error are shown with biological triplicated samples

Fig. 5

Location of upregulated or downregulated genes in AR-expressing HFDPC-K4DT cells in focal adhesion pathway from KEGG

Fig. 6

Location of upregulated or downregulated genes in AR-expressing HFDPCK4DT cell in Proteoglycans in cancer pathway from KEGG