Panoply™ Human EPHA3 Knockdown Stable Cell Line

Eph receptor tyrosine kinase and its ligand Ephrin mediate important cell communications during normal and tumorigenesis and development, playing a central role in a series of processes including angiogenesis, stem cell maintenance, and cancer metastasis. Eph receptor A3 (EphA3) is widely overexpressed in various cancers, including gastric cancer (GC), and is associated with tumor progression. Previous studies have suggested that EphA3 may play an important role in tumorigenesis and angiogenesis in gastric cancer. However, its exact role and mechanism in gastric cancer remain unclear. Here, functional analysis showed that the loss of EphA3 expression inhibited the proliferation and tumorigenicity of HGC-27 cells in vitro and in vivo. Furthermore, knockdown of EphA3 expression in HGC-27 cells inhibited tubular formation and migration of HUVEC endothelial cells. Knockdown of EphA3 in HGC-27 cells also suppressed tumor angiogenesis in vivo and reduced microvessel density (MVD) in a xenograft model. Further research revealed that the absence of EphA3 inhibits tumor angiogenesis and migration through signal transduction and the STAT3/VEGF signaling pathway. These results suggest that EphA3 may be an effective prognostic indicator and a potential target for gastric cancer treatment.

To determine whether EphA3 knockdown affected the proliferation of HGC-27 cells, researchers used CCK-8 and colony formation assays to examine cell growth. The results showed that the proliferation of EphA3-knockdown HGC-27 cells was inhibited compared to the control group (Figure 1A). Furthermore, EphA3 knockdown also reduced colony formation ability (Figure 1B). Researchers further investigated the effect of EphA3 on tumor growth using a mouse model of subcutaneous xenografting of HGC-27 cells. EphA3 knockdown did not induce any toxicity in mice; however, compared to nude mice transplanted with control cells, nude mice transplanted with EphA3-knockdown HGC-27 cells showed a significantly reduced mean tumor volume (39.1%) (Figures 1C and D). These results indicate that the absence of EphA3 expression inhibits the proliferation of HGC-27 cells both in vitro and in vivo.

Figure 1. EphA3 knockdown and tumor growth of HGC-27 cells. (Lv X Y, et al., 2018)