RESUMEN
AIM: To explore the effects of kaempferol on the proliferation, invasion and migration abilities of HBx-HepG2 cells and to examine the underlying molecular mechanisms.METHODS: The expression levels of related genes at mRNA and protein levels were determined by RT-qPCR and Western blot.The cell apoptotic rate was analyzed by flow cytometry.The cell proliferation, growth, invasion and migration abilities were measured by MTT assay, colony formation assay, Transwell invasion assay and wound healing assay, respectively.RESULTS: Kaemferol inhibited HBx-HepG2 cell proliferation in a concentration-and time-dependent manner.Kaempferol at 100 μmol/L significantly inhibited the colony formation, invasion and migration abilities of the HBx-HepG2 cells.Kaemferol at 100 μmol/L also increased cell apoptotic rate, increased the protein levels of cleaved caspase-3, cleaved caspase-9 and Bax, and decreased the expression level of Bcl-2.In addition, kaemferol at 100 μmol/L suppressed the mRNA and protein expression levels of β-catenin, c-Myc and cyclin D1 in the HBx-HepG2 cells.Kaemferol at 100 μmol/L also suppressed the protein level of p-GSK-3β and the β-catenin protein levels in both cytoplasm and nucleus.LiCl treatment reversed the inhibitory effect of kaempferol on the growth, invasion and migration of the HBx-HepG2 cells.CONCLUSION: Kaempferol inhibits cell proliferation, invasion and migration via activating Wnt/β-catenin signaling in HBx-HepG2 cells.
RESUMEN
AIM:To investigate the regulation of miR-222 on BCL2L13 gene and its effect on the growth and apoptosis of HBx-HepG2 cells, and to explore the underlying molecular mechanisms .METHODS:The expression level of miR-222 was detected by RT-qPCR.The HBx-HepG2 cell growth was examined by MTT and colony formation assays .The cell cycle and apoptosis were analyzed by flow cytometry .The recombination vector pmirGLO-BCL2L13 was constructed, and dual-luciferase reporter experiment was performed to validate the target of miR-222.RESULTS:The expression level of miR-222 in the HBx-HepG2 cells was significantly higher than that in the L 02 cells ( P<0.05 ) .Over-expression of miR-222 enhanced HBx-HepG2 cell growth, changed cell cycle, and inhibited apoptosis (P<0.05).Knockdown of miR-222 reduced HBx-HepG2 cell growth, changed cell cycle, and increased cell apoptotic rate (P<0.05).BCL2L13 was down-regulated in the HBx-HepG2 cells as compared with L02 cells (P<0.05), and knockdown of miR-222 in the HBx-HepG2 cells increased the expression level of BCL2L13 (P<0.05).The results of dual-luciferase reporter assay and re-store experiment showed that miR-222 negatively regulated the expression of BCL2L13 via targeting 3’ UTR of BCL2L13, resulting in the promotion of HBx-HepG2 cell growth .CONCLUSION: miR-222 enhances HBx-HepG2 cell growth via down-regulation of BCL2L13.