Figure 1.

Identification of the trp expression signature in PTEC. (a) Heatmap showing the relative mRNA expression of the trp genes in TECs of different origins. TEC expression normalized based on normal endothelial counterparts: PTEC1-3 expression is normalized against HPrMEC expression, while BTEC and RTEC expression is normalized against HMEC and GEC expression, respectively. (b) Differential analysis of PTEC1-3 and HPrMEC highlighted six genes (i.e., trpa1, trpc1, trpc4, trpm1, trpm7, and trpml1) that are differentially deregulated (adjusted p-value < 0.1, red line), while the other genes did not show significant deregulation. (c) Differential analysis identified six genes that are differentially deregulated (panels a,b), and twelve genes displayed at least 2-fold change in their expression in at least two patient lines (panel e). Among these, four channels (trpa1, trpv2, trpc3 and trpc6) were selected as ‘prostate-associated’. (d) Real-time qPCR analysis of mRNA expression shows that trpa1, trpv2, trpc3 and trpc6 are deregulated in PTEC1-3 relative to those in normal HPrMEC, and the deregulation is observed only in the prostatic tissue but not in the other normal ECs or TECs tested. Values are expressed as the mean ΔΔCT values ± SEM of at least three independent experiments. *: p-value < 0.05. (e) Scatter plots showing the expression of trp channels in PTEC1 (left panel), PTEC2 (central panel) and PTEC3 (right panel) relative to that in HPrMEC, as determined by real-time qPCR. Gray dots indicate genes with less than 2-fold change in PTEC relative to those in HPrMEC. Blue dots indicate genes with more than 2-fold change in at least one PTEC line relative to those in HPrMEC. Red dots showing the four ‘prostate-associated’ genes (i.e., trpa1, trpv2, trpc3, and trpc6), with more than 2-fold change in at least 2 PTEC lines out of three. Correlation values for trp channels are expressed as Log (C_(T,gene x)/C_(T,18s)) in primary PTEC (x-axis) and normal HPrMECs (y-axis).