Volume 29, Issue 4, 2020
DOI: 10.24205/03276716.2020.811
Effect of lncRNA LINC00483 on Taxol Resistance of Hepatocellular Carcinoma Cells by Targeting miR-646
Abstract
Objective: To investigate the effects of lncRNA LINC00483 on the proliferation, migration and invasion of hepatocellular carcinoma cells and taxol resistance, as well as the potential molecular mechanism.
Methods: Hepatocellular carcinoma HCCLM3 cells and taxol resistant HCCLM3/Tax cells were treated with taxol (Tax) of different concentrations (0.08 μmol/L, 0.16 μmol/L, 0.32 μmol/L, 0.64 μmol/L, 1.28 μmol/L, 2.56 μmol/L). CCK8 assay was used to determine the proliferation inhibition rate and IC50 of Tax on HCCLM3 cells. qRT-PCR was carried out to detect the levels of lncRNA LINC00483 and miR-646 in HCCLM3 and HCCLM3/Tax cells. Western blot was applied to detect the expression levels of proteins CyclinD1, p21, MMP2 and MMP9. Transwell assay was used to detect the cell migration and invasion ability. Dual-luciferase reporter assay system was performed to validate the relationship between LINC00483 and miR-646.
Results: The inhibition rate of taxol (Tax) of 0.08 μmol/L, 0.16 μmol/L, 0.32 μmol/L, 0.64 μmol/L, 1.28 μmol/L, 2.56 μmol/L on HCCLM3/Tax cells was significantly lower than that of HCCLM3 cells in a concentration-dependent manner. The IC50 (10.70±0.11) μmol/L of HCCLM3/Tax on Tax was significantly higher than that of HCCLM3 cells (0.49±0.04) μmol/L. In HCCLM3/Tax cells, the level of lncRNA LINC00483 was significantly higher than that of HCCLM3 cells (P<0.05), and the level of miR-646 was significantly lower than that of HCCLM3 cells (P<0.05). Inhibition of lncRNA LINC00483 or over-expression of miR-646 combined with 0.16 μmol/L Tax treatment inhibited HCCLM3/Tax cell migration and invasion, enhanced inhibition rate of Tax on cell proliferation, up-regulated the level of p21 protein, and downregulated the level of CyclinD1, MMP2 and MMP9. LncRNA LINC00483 negatively regulated the expression of miR-646 by targeting. Interfering miR-646 reversed the effects of lncRNA LINC00483 inhibition on proliferation, migration, invasion and taxol resistance of HCCLM3/Tax cells.
Conclusions: Inhibition of lncRNA LINC00483 inhibits proliferation, migration and invasion of HCCLM3/Tax cells and reduces the taxol resistance of cells by targeting miR-646. LncRNA LINC00483 is a potential molecular target for hepatocellular carcinoma.
Keywords
Hepatocellular carcinoma, lncRNA LINC00483, miR-646, Proliferation, Migration, Invasion, Taxol resistance.