[Mechanism of flavonoids of Sophorae Fructus in inhibiting proliferation, migration and invasion of hepatocellular carcinoma cells by regulating LncRNA FBXL19-AS1/miR-342-3p pathway].

Abstract

The aim of this paper was to investigate the effect of flavonoids of Sophorae Fructus on the proliferation, migration and invasion of hepatocellular carcinoma cells and analyze the regulatory mechanism of LncRNA FBXL19-AS1/miR-342-3 p pathway. MTT assay and plate cloning assay were used to detect the effect of flavonoids of Sophorae Fructus at different concentrations(1, 5, 10 mg·mL~(-1)) on the proliferation of liver cancer Huh7 cells. The effect of flavonoids of Sophorae Fructus on the migration and invasion of Huh7 cells was examined by Transwell chamber assay. qRT-PCR was used to detect the effect of flavonoids of Sophorae Fructus on the expression levels of FBXL19-AS1 and miR-342-3 p in Huh7 cells. The dual luciferase reporter assay was used to detect whether FBXL19-AS1 targeted at miR-342-3 p. The effect on the inhibition of FBXL19-AS1 expression or FBXL19-AS1 overexpression and then the proliferation, migration and invasion of Huh7 cells were examined by the above methods. Gelatin zymography was used to detect the activities of MMP-2 and MMP-9. The expression levels of cyclinD1, p21, MMP-2 and MMP-9 proteins were detected by Western blot. Flavonoids of Sophorae Fructus significantly inhibited the proliferation, migration and invasion of Huh7 cells(P<0.05), promoted the expression of p21 protein(P<0.05), and inhibited the expressions of cyclinD1, MMP-2 and MMP-9(P<0.05) in a dose-dependent manner, and could reduce the activity of MMP-2 and MMP-9(P<0.05). The expression level of FBXL19-AS1 was significantly decreased in Huh7 cells treated with flavonoids of Sophorae Fructus(P<0.05), whereas the expression level of miR-342-3 p was significantly increased(P<0.05). The dual luciferase reporter assay confirmed that FBXL19-AS1 targeted at the inhibition of miR-342-3 p expression. After inhibiting the expression of FBXL19-AS1, the inhibition rate of cell proliferation was significantly increased(P<0.05), the number of cell clone formation was significantly reduced(P<0.05), the number of migrated cells and the number of invasive cells were significantly decreased(P<0.05), and the expression levels of cyclinD1, MMP-2 and MMP-9 were significantly decreased(P<0.05), the activities of MMP-2 and MMP-9 were significantly reduced(P<0.05), while the expression level of p21 protein was significantly increased(P<0.05). The overexpression of FBXL19-AS1 reversed the inhibitory effect of flavonoids of Sophorae Fructus on the proliferation, migration and invasion of Huh7 cells. Flavonoids of Sophorae Fructus could inhibite the proliferation, migration and invasion of hepatoma cells by regulating LncRNA FBXL19-AS1/miR-342-3 p pathway.