Effects of theaflavins on HepG2 cell proliferation and apoptosis by regulating Wnt/β-catenin and Hedgehog signaling pathway
Objective To investigate the effects of theaflavins on the proliferation,apoptosis and Wnt/β-catenin and Hedgehog signaling pathway in hepatocellular carcinoma HepG2 cells.Methods HepG2 cells were treated with different concentrations of theaflavins(0,2.5,5,10,20,40,80,160 and 320 μmol/L)and cultured for 24,48 or 72 hours.The viability of HepG2 cells was detected by CCK-8 assay.According to the results of HepG2 cell viability experiment,the subsequent experiments were divided into blank control group,low dose of theaflavins(20 μmol/L)group,middle dose of theaflavins(40 μmol/L)group and high dose of theaflavins(80 μmol/L)group,and HepG2 cells were cultured for 24 hours.Cell proliferation was detected by colony formation assay and cell apoptosis was detected by flow cytometry.The levels of Ki67,PCNA,cleaved caspase-3,cleaved caspase-9,GLi1,SMO,PTch1,β-catenin,c-myc and cyclin D1 protein were detected by western blot.The levels of GLi1,SMO,PTch1,β-catenin,c-myc and cyclin D1 mRNA expression were detected by qRT-PCR.Results Compared with the control group,the HepG2 cell activity of theaflavins group was decreased,the cell apoptosis rate was increased,and the levels of GLi1,SMO,PTch1,β-catenin,c-myc and cyclin D1 mRNA expression were significantly downregulated(P<0.05).The levels of Ki67,PCNA,GLi1,SMO,PTch1,β-catenin,c-myc and cyclin D1 protein were downregulated,while the levels of cleaved caspase-3 and cleaved caspase-9 protein were significantly upregulated in theaflavins group(P<0.05).Conclusion Theaflavins could inhibit HepG2 cell proliferation and promote cell apoptosis by regulating the Wnt/β-catenin and Hedgehog pathway.
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