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Archive > Volume 29 Issue 7 > 2022,29(7):639-645. DOI:10.3872/j.issn.1007-385X.2022.07.005 Prev Next
Basic Research
The effects of miR-1243 on the proliferation and migration of hepatocellular carcinoma HepG2 cells through targeted regulation of hnRNPA2B1 expression
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Abstract:
Objective: To investigate the effects of miR-1243 on the proliferation and migration of hepatocellular carcinoma HepG2 cells by targeting and regulating the expression of heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2B1) and its molecular mechanisms. Methods: qPCR and WB methods were used to detect the levels of miR-1243 and hnRNPA2B1 mRNA and the expression of hnRNPA2B1, cyclin D1 and matrix metalloproteinase-2 (MMP-2) protein in 40 HCC tissues and adjacent tissues (surgically removed in Shouyi Hospital of Wuhan Third Hospital from January 2019 to August 2021), human normal liver QSG-7701 cells and hepatocellular carcinoma HepG2, Hep3b and Huh-7 cells. Dual-luciferase reporter assay was performed to verify the targeting relationship between miR-1243 and hnRNPA2B1. HepG2 cells were randomly divided into control group (not transfected), miR-NC group (transfected with miR-NC), miR-1243 mimic group (transfected with miR-1243 mimic), miR-1243 mimic+pcDNA3.1 group (transfected with miR-1243 mimic and pcDNA3.1), and miR-1243 mimic+pc-hnRNPA2B1 group (transfected with miR-1243 mimic and pc-hnRNPA2B1) and corresponding transfections were then carried out. CCK-8 method was performed to evaluate the proliferation ability of HepG2 cells in each group after transfection. Scratch healing test was performed to evaluate the migration ability of HepG2 cells, and Western blotting was performed to determine the protein expression levels of hnRNPA2B1, cyclin D1 and matrix metalloproteinase-2 (MMP-2). Results: Compared with adjacent tissues or human normal liver cells QSG-7701, the expression level of miR-1243 in liver cancer tissues and cells was significantly reduced, and the expression levels of hnRNPA2B1 mRNA and protein were significantly increased (all P<0.05). The results of Dual-luciferase experiments showed that miR-1243 and hnRNPA2B1 had a targeting relationship, and miR-1243 targeting negatively regulated the expression of hnRNPA2B1 protein. After miR-1243 mimic transfection,the expression of hnRNPA2B1 protein in HepG2 cells, proliferation ability of cells, scratch healing rate, and the expression of cyclin D1 and MMP-2 proteins were all significantly decreased (all P<0.05); the overexpression of both hnRNPA2B1 and miR-1243 could reverse the inhibitory effects of miR-1243 overexpression on the proliferation and migration of HepG2 cells. Conclusion: miR-1243 inhibits the proliferation and migration of liver cancer cells through targeting the expression of hnRNPA2B1.
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