Abstract
BackgroundHepatocellular carcinoma (HCC) is one of the most common malignant tumors of the digestive system, with a low 5-year survival rate (5–9%). The abnormal expression of miRNA in liver cancer cells may play an important role in the pathophysiology of liver cancer. The ability of tumor invasion and metastasis is an important indicator to evaluate the degree of malignancy of HCC. Autophagy may affect the ability of tumor cells to invade and metastasize. Autophagy-related genes and proteins (Atg) are the core and key to regulating autophagy. The purpose of this study was to investigate the effect of microRNA-7 (miR-7) on targeting autophagy-related protein Atg5 to inhibit the effect of autophagy on the invasion and metastasis ability of liver cancer cells.MethodsThe content of miR-7 and Atg5 in normal liver tissue and HCC tissues was detected by quantitative real-time polymerase chain (qRT-PCR). SMMC-7721 hepatoma cells were cultured in vitro, a starvation environment was simulated to activate autophagy, and transfection of cells was carried out by using miR-7 mimic, inhibitor, and autophagic inhibitor 3-MA. The RFP-GFP-LC3 double-labeled adenovirus infected hepatoma cells, and autophagy was detected by fluorescence microscopy. Western blot was used to detect the expression of LC3, Atg5, and the epithelial-mesenchymal transition (EMT)-related proteins (N-cadherin, vimentin, and snail). Transwell and plate cloning were used to detect the proliferation, invasion, and metastasis of hepatocarcinoma cells.ResultsExpression of miR-7 (16.72±4.71, P<0.05) in HCC tissues was low, but the expression of Atg5 (13.70±2.80, P<0.05)was high. MiR-7 and Atg5 were highly negatively correlated in hepatoma tissues (r=−0.97). With the overexpression of hepatoma cells, the expression of Atg5 (0.49±0.07, F=395.26) and LC3II (0.51±0.06, F=23.58) was increased (P<0.05), and the autophagy was enhanced. As a result, the proliferation of hepatocarcinoma cells was decreased (t=3.22, P<0.05), the expression of EMT-related protein [N-cadherin (0.37±0.04), vimentin (0.60±0.07), snail (0.54±0.07)] was decreased (P<0.05), and hepatoma cell invasion and metastasis were decreased (n=6, F=162.28, P<0.05).ConclusionsMiR-7 can inhibit the invasion and metastasis of hepatoma cells by targeting Atg5 to regulate autophagy.
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