Abstract
This study aims at exploring the regulatory effects of miR-193a-5p on hepatocellular carcinoma (HCC). It might provide new insight into the improvement of clinical treatment of HCC. A total of 50 HCC patients who did not receive any tumor treatments were recruited, and 50 paired tumor tissues and adjacent non-tumor tissues were obtained. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to validate the expression and significance of miR-193a-5p in HCC tumor tissues, adjacent non-tumor tissues and cell lines. Binding-site of the target gene of miR-193a-5p was predicted by bioinformatics and further verified by Dual-Luciferase reporter gene assay and Western blotting (WB) assay. To investigate the potential role of miR-193a-5p in HCC development, cell counting kit-8 (CCK-8) assay was performed to study the proliferation and viability capacities. Flow cytometric analyses were adopted to test the cell cycle distribution and quantify the apoptotic cell proportion. MiR-193a-5p expression was specifically up-regulated in HCC tissues and cell lines compared with paired adjacent non-tumor tissues and normal liver cell lines (HL-7702) respectively. BMF was considered as a downstream gene of miR-193a-5p, which was further proofed in Dual-Luciferase reporter gene assay and Western blot assays. In vitro experiments showed that miR-193a-5p overexpression could accelerate the proliferation, facilitate the G1/S transition and suppress the apoptosis of HCC cells. However, BMF overexpression could reverse the effects of miR-193a-5p on the cellular functions of HCC cells. This finding suggested that miR-193a-5p is strongly up-regulated in HCC. MiR-193a-5p promoted the abnormal proliferation of HCC cells and limited their apoptosis by targeting the downstream gene BMF. Thus, the miR-193a-5p/BMF axis might be a novel regulatory pathway of apoptosis which could be potential therapeutic sites in HCC treatment.
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