Panoply™ Human EPHB4 Knockdown Stable Cell Line

Erythropoietin producing human hepatoma (EphB4) promotes the survival, migration, invasion, and angiogenesis of tumor cells from various sources. Here, researchers investigated the effects of EphB4 on the oncogenic potential of renal cell carcinoma cells, including in vitro and in vivo experiments. They used CCK-8 and Annexin V-FITC/PI staining methods to detect the effects of EphB4 knockdown on cell proliferation and cell death. The Boyden chamber assay and wound healing assay were used to detect the effects of EphB4 knockdown on cell invasion and migration. To explore the downstream effects of EphB4 knockdown, the researchers examined the activities of ERK and STAT3. The results showed that EphB4 downregulation inhibited the growth, invasion, and migration of renal cell carcinoma cells and promoted apoptosis. EphB4 knockdown also reduced the levels of p-ERK and p-STAT3. In vivo experiments showed that EphB4 knockdown significantly inhibited the growth of xenograft tumors and reduced the levels of p-ERK and p-STAT3. Therefore, EphB4 knockdown can effectively inhibit the proliferation of human renal cell carcinoma cells and induce their apoptosis both in vitro and in vivo.

Here, researchers overexpressed EphB4, resulting in increased RCC cell proliferation (detected by CCK-8 assay) (Figure 1A-D). Similarly, in EphB4 knockdown cells, cell proliferation was significantly reduced (Figure 1A-D). These results indicate that EphB4 positively regulates cell proliferation. Researchers used Annexin V-FITC/PI double staining to detect the apoptotic effect of EphB4 on RCC cells. Flow cytometry analysis was performed immediately after staining. The results showed that EphB4 downregulation promoted apoptosis. Annexin V-FITC staining revealed early apoptosis in EphB4 knockdown 786-O cells (Figure 1E, 1F). Compared to the control group, the apoptosis rate in EphB4 knockdown cells was significantly increased.

Figure 1. Effect of EphB4 on proliferation and apoptosis of RCC cells. (Zhang Y, et al., 2019)