Panoply™ Human CXCL13 Knockdown Stable Cell Line

C-X-C motif chemokine ligand 13 (CXCL13) belongs to the CXC subfamily of chemokines and influences the biological processes of various cell types as well as the progression of numerous clinical diseases. Here, researchers elucidated the underlying mechanisms linking CXCL13 to benign prostatic hyperplasia (BPH). CXCL13 is highly expressed in prostate tissue and is upregulated in BPH. The study found that in BPH-1 cells, CXCL13 regulates cell proliferation, apoptosis, and epithelial-mesenchymal transition (EMT) via the CXCR5 receptor and the AKT and ERK1/2 signaling pathways; meanwhile, in WPMY-1 cells, CXCL13 promotes inflammation and fibrosis via the CXCR5 receptor and the STAT3 signaling pathway. In vivo experiments demonstrated that rHuCXCL13 induced BPH in rats. Furthermore, CXCL13 levels positively correlated with prostate volume and total prostate-specific antigen (tPSA) levels. These data indicate that CXCL13 regulates cell proliferation, the cell cycle, and epithelial EMT-mediated by various inflammatory factors-and induces fibrosis in prostatic stromal cells, suggesting that CXCL13 is a promising therapeutic target for BPH.

Here, researchers generated CXCL13-knockdown BPH-1 cells and verified the knockdown efficiency via qRT-PCR and Western blot (Figure 1A-C). Flow cytometry results demonstrated that CXCL13 knockdown significantly promoted apoptosis (Figure 1D, E), induced G0/G1 phase arrest (Figure 1F, G), and reduced cell proliferation (Figure 1H). Western blot analysis revealed elevated levels of the pro-apoptotic protein BAX and significantly reduced levels of the anti-apoptotic protein BCL-2 in CXCL13-knockdown cells (Figure 1I, K). Concurrently, the levels of proteins regulating the G0/G1 phase (Cyclin D1, CDK2, CDK4) were significantly decreased (Figure 1I, K). Furthermore, CXCL13 knockdown resulted in significantly reduced levels of pAKT and pERK1/2, while total AKT and ERK1/2 protein levels remained unchanged (Figure 1J, K). Additionally, CXCL13 knockdown inhibited the epithelial-mesenchymal transition (EMT) process, evidenced by increased E-cadherin expression and decreased N-cadherin expression (Figure 1L, M). However, treatment with 100 ng/mL rHuCXCL13 almost completely reversed the effects of CXCL13 knockdown on BPH-1 cell proliferation, apoptosis, and EMT (Figure 1).

Figure 1. Knockdown of CXCL13 inhibited proliferation, EMT and promoted apoptosis of BPH-1 cells via ERK1/2 and AKT pathway. (Liu D, et al., 2022)