Expression and regulation network of BLNK in CP/CPPS in animal, cell model and clinical samples.

Abstract

B-cell linker (BLNK) is associated with the inflammatory process of certain diseases. However, no studies have explored the role of BLNK in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). We performed transcriptome analysis of the rat model and found that BCR signaling pathway, especially the BLNK gene, is closely related to CP/CPPS. We detected the expression levels of BLNK gene or protein in CP/CPPS rat tissues, human expressed prostatic secretion (EPS), and cell models by qPCR, WB, and IHC. Subsequently, we silenced the BLNK gene in human prostatitis cell model. The cell cycle, and expression level of inflammatory factors were detected; the core pathway and interacting molecules of BLNK were revealed by mRNA/lncRNA sequencing, bioinformatics analysis and validation. Results showed that the BLNK gene or protein was significantly upregulated in the CP/CPPS rat tissues, human EPS, and cell model. Functionally, BLNK silencing inhibited the proliferation, promoted cell apoptosis and S phase arrest, and inhibited the expression of IL-1β, IL-6, and IL-8 at the human prostatitis cell model. Furthermore, the DEGs and DE-lncRNA induced by BLNK silencing were significantly enriched in the calcium signaling pathway, the BLNK has strongly interacted with EDNRB, FGF1, FGF2, and F2R. Moreover, in prostatitis group, all four genes are target genes of LINC02518, and their expression levels were significantly altered after BLNK silencing. These results suggest the important role of BLNK in CP/CPPS. BLNK may affect the proliferation and inflammatory factor secretion of CP/CPPS through calcium signaling pathway, which may be closely related to LINC02518.