Panoply™ Human TNFRSF11B Knockdown Stable Cell Line

Tumor necrosis factor receptor superfamily member 11B (TNFRSF11B) has been implicated in the development and progression of various human malignancies. However, the complex mechanisms underlying TNFRSF11B in human gastric cancer (GC) remain largely unknown. Here, researchers evaluated the expression and clinical significance of TNFRSF11B in 70 and 160 GC tissues using immunohistochemistry and gene chip analysis, respectively. The biological functions of TNFRSF11B were investigated using in vitro and in vivo experiments. Results showed that TNFRSF11B was highly expressed in the cytoplasm of GC cells and correlated with poor patient prognosis. TNFRSF11B significantly promoted GC cell proliferation, migration, and invasion, as well as tumorigenesis in vitro and in vivo. Furthermore, TNFRSF11B inhibited GC cell apoptosis. Nuclear β-catenin expression was positively correlated with TNFRSF11B expression. TNFRSF11B directly binds to GSK-3β, promoting its phosphorylation and thereby upregulating the expression of β-catenin and its downstream signaling molecules. In summary, TNFRSF11B promotes the malignant phenotype of gastric cancer cells and activates the Wnt/β-catenin signaling pathway. Therefore, TNFRSF11B is expected to become a biomarker for gastric cancer, and inhibiting TNFRSF11B expression may provide a new therapeutic target for gastric cancer patients.

Here, researchers examined the role of TNFRSF11B in proliferation by using the RTCA system. The results indicated that the proliferation activities were strongly promoted in TNFRSF11B-overexpressing HGC-27 and BGC-823 cells (Figure 1A and B). The proliferation activities were significantly inhibited in TNFRSF11B-knockdown MGC-803 and SGC-7901 cells (Figure 1C and D). Furthermore, a colony formation assay was performed to further investigate the regulatory role of TNFRSF11B in the clonogenicity of gastric cancer cells. Upregulation of TNFRSF11B significantly enhanced the clonogenicity of HGC-27 and BGC-823 cells (Figure 1E and F), whereas downregulation of TNFRSF11B significantly reduced the clonogenicity of MGC-803 and SGC-7901 cells (Figure 1G and H). To assess the tumorigenic potential of TNFRSF11B, control and TNFRSF11B-knockdown MGC803 cells were subcutaneously injected into mice. Five mice in each experimental and control group were observed for two months and then sacrificed. The results showed that knockdown of TNFRSF11B in MGC803 cells significantly reduced the volume and weight of transplanted tumors, and tumor growth in the shTNFRSF11B group was significantly slower than that in the control group (Figure 1I). These in vitro and in vivo results suggest that TNFRSF11B plays an important role in gastric cancer progression.

Figure 1. TNFRSF11B promotes GC cell growth in vitro and in vivo. (Luan F, et al., 2020)