Objective To investigate the molecular mechanism of shikonin in inhibiting the proliferation of hepatocellular carcinoma(HCC)cells.Methods HepG2 human HCC cells were divided into six groups:control,low-dose shikonin,high-dose shikonin,miR-181c negative control(NC),miR-181c-inhibitor,and miR-181c-mimic groups.Cells were treated respectively with 2.5 or 5.0 μmol/L concentrations of shikonin,and transfected with miR-181c inhibitor or miR-181c mimics,respectively.Cell proliferation was detected using the cell counting kit-8(CCK-8)assay,while apoptosis was assessed by flow cytometry.The relative mRNA expression levels of miR-181c and non-SMC condensin I complex subunit G(NCAPG)were measured by qRT-PCR,and the relative protein expression levels of NCAPG and B-cell lymphoma 2(Bcl-2)were determined by Western blot.Results Compared with the control group,the cell proliferation rate was significantly reduced in both the high-dose and low-dose shikonin groups(both P＜0.05),and that of the high-dose group reduced more remarkably than the low-dose group(P＜0.05).In comparison to the miR-181c NC group,the cell proliferation rate significantly increased in the miR-181c inhibitor group,while significantly decreased in the miR-181c mimic group(both P＜0.05).The high-dose group exhibited a significantly higher apoptosis rate than both the low-dose group and control group(both P＜0.05).Compared with the miR-181c NC group,the apoptosis rate significantly decreased in the miR-181c inhibitor group,while significantly increased in the miR-181c mimic group(both P＜0.05).The relative expression level of miR-181c mRNA in the high-dose group was significantly higher than that of the low-dose and control groups(both P＜0.05).Compared to the miR-181c NC group,the miR-181c mRNA relative expression level significantly decreased in the miR-181c inhibitor group,while significantly increased in the miR-181c mimic group(both P＜0.05).The relative expression levels of NCAPG mRNA,NCAPG and Bcl-2 proteins in the high-dose group were significantly lower than those of the low-dose group and control group(all P＜0.05).Compared to the miR-181c NC group,the relative expression levels of NCAPG mRNA,NCAPG and Bcl-2 proteins significantly increased in the miR-181c inhibitor group,while significantly decreased in the miR-181c mimic group(all P＜0.05).Conclusion Shikonin can upregulate miR-181c expression in HCC cells,thereby inhibiting the proliferation of HCC cells.
维普
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