HIF-1α promotes ZEB1 expression and EMT in a human bladder cancer lung metastasis animal model.

Lung is one of the most common sites for bladder cancer to metastasize. Although the involvement of the epithelial-to-mesenchymal transition (EMT) in bladder cancer progression has been established, the mechanism of EMT induction remains unclear. In order to investigate this, T24-parental (P) and T24-lung (L) bladder cancer cells were obtained from primary tumors and lung metastatic sites of an animal model with orthotopic spontaneous metastatic bladder cancer, according to a protocol previously described. Compared with T24-P cells, mesenchymal-like T24-L cells exhibited an increased ability in tumor invasion and metastasis, as well as an increased expression of hypoxia-inducible factor (HIF)-1 alpha, zinc finger E-box-binding homeobox 1 (ZEB1), vimentin and N-cadherin and lower level of cytokeratin 18 were observed. Mechanistically, it was identified that HIF-1 alpha increases ZEB1 expression and subsequently regulates the expression of EMT-related genes in both HIF-1 alpha knocking down by siRNA and gain-in HIF-1 alpha by hypoxia culture cell models. In addition, the expression of HIF-1 alpha and ZEB1 in bladder cancer tissues were increased compared with normal bladder epithelial tissues, as well as significantly increased in the high-grade, invasive and metastatic bladder cancer tissues compared with low-grade, superficial and non-metastatic bladder cancer tissues by using immune-histochemical staining assay. Notably, the protein level of HIF-1 alpha was positively associated with that of ZEB1 in bladder cancer tissues. Results from the present study indicate that HIF-1 alpha promotes ZEB1 expression and EMT in the T24-L human bladder cancer lung metastasis animal model, suggesting that HIF-1 alpha serves an important function in the metastasis of bladder cancer, and HIF-1 alpha and ZEB1 may be potential targets for inhibiting bladder metastasis in the future.