Liver regeneration phosphatase 3 promotes angiogenesis in colorectal cancer cells via upregulating hypoxia inducible factor 1-α

Abstract

Objective To investigate whether the liver regeneration phosphatase 3 (PRL-3) upregulates the expression of hypoxia inducible factor 1-α (HIF1-α), promoting angiogenesis. Methods LoVo cells were selected for constructing PRL-3-overexpression lines, and HT29 cells were chosen for the knockdown groups. We performed qPCR and Western blotting to identify the expression of HIF1-α and VEGF in cell lines mentioned above compared with their negative control group respectively. Besides, their supernatants were collected and used to carry out angiogenesis assay in human umbilical vein endothelial cells (HUVECs). In addition, immunohistochemistry was used to confirm the status of HIF1-α and VEGF in xenografts of mice injected with cells mentioned above.Statisticalanalysis of data using the statistical product and service solutions 15.0 software. Results Western blotting and qPCR showed that overexpression of PRL-3 corelated with high levels of HIF1-α [LoVo-P vs. LoVo-NC: 4] and VEGF [LoVo-P vs.LoVo-NC: 5.8] in colorectal cancer cells. Similarly, knocking down PRL-3 showed the opposite trend [HT29-P vs.HT29-NC, HIF1-α: 0.3, VEGF: 0.2, P<0.05]. Also, the angiogenesis assay presented that PRL-3 promoted the formation of new vessels [LoVo-P/LoVo-NC: 8±2 vs. 2±1, HT29-P vs. HT29-NC: 3±1 vs. 11±3 (P<0.05)]. Results of immunohistochemistry were consistent with those in Western blotting and qPCR[ LoVo-P vs. LoVo-NC, HIF1-α: 89% vs. 33%, VEGF: 83% vs. 25%, HT29-P/HT29-NC, HIF1-α: 8% vs. 67%, VEGF: 12% vs. 82%, P<0.05]. Conclusion PRL-3 in colorectal cancer cells promotes angiogenesis by increasing the expression of HIF1-α. Key words: Colorectal cancer; Angiogenesis; Liver regeneration phosphatase 3; Hypoxia inducible factor 1-α