The regulatory mechanism of miR-370 on the proliferation and migration of hepatocellular carcinoma cells via targeting RNA binding protein FUS

Abstract

Abstract: Objective To investigate the mechanism by which miR-370 targets FUS to regulate the proliferation and migration of hepatocellular carcinoma (HCC) cells. Methods A vector named pcDNA3/pri-miR-370 for overexpression miR-370 was synthesized and constructed, and the corresponding antisense oligonucleotide ASO-370 was prepared. MiRNA was amplified using bacterial transformation and subsequently transfected into HCC cell lines to investigate its effect on cellular functions. The experiment was divided into four groups: pcDNA3 empty vector group (pcDNA3 group), pcDNA3/pri-miR-370 plasmid group (miR-370 group), scrambled oligonucleotide group (ASO-ctrl group), and miR-370 ASO group (ASO-370 group). Cell proliferation ability was assessed using the CCK-8 assay; cell migration and invasion capabilities were evaluated using Transwell assays. Bioinformatics analysis was performed to predict the candidate target gene of miR-370, the RNA-binding protein FUS. The expression levels of FUS at both mRNA and protein levels in miR-370 overexpressing HCC cell lines were measured by real-time quantitative PCR and Western blotting. Results After overexpressing miR-370, the proliferative ability of HepG2 liver cancer cells were significantly inhibited, while inhibiting the expression of miR-370 in the cells enhanced their proliferative ability. The relative migration numbers of the pcDNA3 group, miR-370 group, ASO-ctrl group, and ASO-370 group were (1.13±0.24), (0.49±0.13), (1.37±0.31), and (1.58±0.39), respectively. The FUS mRNA expression levels in the pcDNA3 group, miR-370 group, ASO-ctrl group, and ASO-370 group were 1, (0.56±0.08), 1, and (3.62±1.51), respectively. The FUS mRNA expression level in the miR-370 group was lower than that in the pcDNA3 group, while the FUS mRNA expression level in the ASO-370 group was higher than that in the ASO-ctrl group. The relative expression levels of FUS protein in the pcDNA3 group, miR-370 group, ASO-ctrl group, and ASO-370 group were 1, (0.59±0.12), 1, and (1.38±0.29), respectively. The FUS protein expression level in the miR-370 group was lower than that in the pcDNA3 group, while the FUS protein expression level in the ASO-370 group was higher than that in the ASO-ctrl group. The differences were statistically significant (P<0.05). Conclusion miR-370 regulates the proliferation and migration of HCC line HepG2 by inhibiting the expression of the RNA-binding protein FUS, indicating that miR-370 may serve as a viable therapeutic target for suppressing HCC.

Key words: miR-370, Hepatocellular carcinoma cells, FUS protein, HepG2

WANG Mi-si, MA Li-tao, ZHANG Li-na, WANG Bin. The regulatory mechanism of miR-370 on the proliferation and migration of hepatocellular carcinoma cells via targeting RNA binding protein FUS[J]. Chinese Hepatolgy, 2026, 31(1): 35-38.

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