ABSTRACT: To date, the role of hematopoietic-substrate-1-associated protein X-1 (HAX1) in liver cancer is rarely studied. The present study explored the role of HAX1 in liver cancer. The association between HAX1 expression and survival of patients with liver cancer was analyzed by a log-rank test. The target genes for HAX1 was predicted by TargetScan and verified by a dual-luciferase reporter assay. The protein and mRNA expressions of HAX1 in liver cancer and adjacent non-cancerous tissues were examined by immunohistochemistry and reverse transcription-quantitative PCR (RT-qPCR). The transfection efficiency of HAX1, small interfering RNA against HAX1, microRNA (miR)-125a mimics, miR-125a inhibitor, miR-223 mimics and miR-223 inhibitor in liver cancer cells was determined by RT-qPCR. The expression of HAX1, p53, VEGF, epithelial-to-mesenchymal transition (EMT)-related markers (E-cadherin, N-cadherin and vimentin) in the cancer cells were determined by western blotting and RT-qPCR. Cell viability, migration, invasion and colony formation rates were determined by Cell Counting Kot-8, wound healing, Transwell and colony formation assays, respectively. The results showed that high expression of HAX1 in liver cancer was found relate to poor prognosis in patients with liver cancer, and upregulation of HAX1 expression in liver cancer tissues was related to lower overall survival. miR-125a-5p directly binds to HAX1. Upregulation of miR-125a-5p expression inhibited cell viability, migration, invasion and colony formation of SK-Hep1 cells and reduced the expression of HAX1, VEGF, N-cadherin and vimentin, but increased cell apoptosis and the expression of p53 and E-cadherin. However, the effects miR-125a-5p upregulation were partially reversed by SK-Hep1 cells with HAX1 overexpression. Downregulated miR-125a-5p in SNU-387 cells produced opposite effects, which was partially reversed by HAX silencing. In conclusion, miR-125a-5p suppresses liver cancer growth via targeting HAX1 and concurrently modulating the expression of p53 and VEGF and EMT-related markers.
