Role of miR-498 Combined with CREB1 in Apoptosis and Invasion of Hepatoma Cell Line.

Abstract

Objective To detect the expression levels of miR-498 in the hepatoma cells and to clarify the biological roles of miR-498 in hepatoma by investigating CREB1, which is the target of miR-498. This study provides a new biomarker for the early diagnosis and targeted therapies for hepatoma. Methods The expression of miR-498 between hepatoma cells and hepatocytes was detected by qRT-PCR. miR-498 was overexpressed in hepatoma cells, and then, flow cytometry was used to analyze the cell apoptosis rate. Cell migration and invasion ability were evaluated by Transwell migration assay and Matrigel invasion assay. The downstream targets of miR-498 were searched in the biological database or related software, and the result can be verified by luciferase reporter assay. The knockdown of the downstream target using RNA interference detected its biological functions in hepatoma cells and was confirmed by cotransfection experiments. Results miR-498 was downregulated in hepatoma cell lines compared with hepatocytes. The overexpression of miR-498 significantly promoted apoptosis. Luciferase reporter assays showed that miR-498 could target CREB1 3′UTR and CREB1 was one of the targets of miR-498. Knockdown of CREB1 also inhibited hepatoma cells' malignant potential and increased the apoptosis rate of hepatoma cells. CREB1 was able to alleviate the changes caused by miR-498 overexpression. Conclusions miR-498 is downregulated in hepatoma cell lines. Therefore, miR-498 can be one of the potential molecular markers for hepatoma diagnosis. miR-498 plays a role in tumor suppression through regulating CREB1.