Panoply™ Human TNFRSF1B Knockdown Stable Cell Line

Androgenetic alopecia (AGA) is a common type of hair loss whose pathogenesis remains incompletely understood. Here, researchers investigated the association between the tumor necrosis factor receptor superfamily member 1B (TNFRSF1B) gene and AGA to validate its potential pathogenic role. RNA sequencing data demonstrated that TNFRSF1B, PIK3CD, and THEMIS2 were significantly upregulated in hair follicles of AGA patients. Receiver operating characteristic (ROC) curve analysis indicated that these three genes had a certain diagnostic value. Weighted gene co-expression network analysis (WGCNA) and gene intersection analysis revealed a strong association between TNFRSF1B and AGA. Mendelian randomization analysis further confirmed the causal relationship between TNFRSF1B and AGA. Meta-analysis also confirmed the upregulation of TNFRSF1B expression in multiple AGA datasets. In vitro experiments demonstrated that TNFRSF1B knockdown enhanced the proliferation and migration of human dermal papilla cells (HDPCs), reduced cell apoptosis, and upregulated the expression of β-catenin and CyclinD1. Furthermore, under conditions of oxidative stress, knockdown of TNFRSF1B reduced reactive oxygen species (ROS) levels and upregulated the expression of superoxide dismutase 2 (SOD2) and catalase (CAT). In vivo experiments showed that knockdown of TNFRSF1B promoted hair growth and alleviated oxidative stress in an AGA mouse model. These results suggest that TNFRSF1B may be a potential pathogenic factor in AGA and provide a new target for its treatment.

To further investigate the role of TNFRSF1B in AGA, researchers generated HDPC cell lines expressing TNFRSF1B overexpression, knockdown, and shRNA-resistant mutations (Figure 1A). Subsequently, cell proliferation was assessed using a CCK-8 assay. As shown in Figure 1B, cell proliferation was significantly increased in TNFRSF1B knockdown cells, whereas it was significantly decreased in TNFRSF1B overexpression cells, indicating that TNFRSF1B inhibits HDPC proliferation. This finding was further confirmed by Ki67 immunofluorescence staining, which showed an increase in the number of Ki67-positive cells in the knockdown group and a decrease in the overexpression group (Figure 1C, D). Furthermore, apoptosis was assessed using a TUNEL assay. As shown in Figure 1E, F, apoptosis was significantly decreased in TNFRSF1B knockdown cells, whereas it was significantly increased in TNFRSF1B overexpression cells, indicating that TNFRSF1B knockdown reduces HDPC apoptosis. Cell migration assays were also performed. As shown in Figure 1G, H, TNFRSF1B gene knockdown significantly enhanced cell migration ability, while TNFRSF1B gene overexpression significantly inhibited cell migration, indicating that TNFRSF1B negatively regulates the migration ability of HDPCs.

Figure 1. Effects of TNFRSF1B on HDPC proliferation, apoptosis, and migration. (Xu F, et al., 2025)