甘草查耳酮A通过miR-1270/ADAM9通路抑制肝癌细胞恶性表型的机制研究OA

Objective:To investigate the effect and mechanism of licochalcone A(Lico A)on the malignant phenotype of hepatocellular carcinoma(HCC)cells.Methods:LO2 and HepG2 cells were treated with culture medium containing 0,5,10,20,40,80,and 160 μmol/L Lico A to screen for the effective concentration of Lico A.HepG2 cells were assigned into control group(normal culture),Lico A group(treated with 20 μmol/L Lico A for 24 hours),Lico A+negative control inhibitor transfection group(treated with 20 μ mol/L Lico A for 24 hours after transfection of negative control inhibitor),Lico A+miR-1270 inhibitor transfection group(treated with 20 μ mol/L Lico A for 24 hours after transfection of miR-1270 inhibitor),Lico A+mimics NC group(Transfect mimics NC and treat with 20 μ mol/L Lico A for 24 hours),Lico A+miR-1270 mimics group(Transfect miR-1270 mimics and treat with 20 μ mol/L Lico A for 24 hours).RT-qPCR was applied to detect the expression of miR-1270 and A Disintegrin and Metalloproteinase 9(ADAM9)mRNA in cells.Cell Connting Kit-8(CCK-8)method was applied to detect cell proliferation.Flow cytometry was applied to detect cell apoptosis.Scratch healing experiment was applied to detect cell migration ability.Transwell chamber was used to detect cell invasion ability.Western blot was applied to detect the expression of ADAM9 protein in cells.Dual luciferase reporter gene experiment was applied to verify the targeting relationship between ADAM9 and miR-1270.Results:20 μmol/L Lico A could reduce the viability of HepG2 cells and had no great toxicity to LO2 cells.After treatment with Lico A and miR-1270 mimics,the expression level of miR-1270 and apoptosis rate in HepG2 cells increased,while the mRNA and protein levels of ADAM9,cell proliferation activity,scratch healing rate,and number of invasive cells decreased(all P＜0.05).Knocking down miR-1270 expression can weaken the inhibitory effect of Lico A on the malignant phenotype of HepG2 cells;Upregulation of miR-1270 expression leads to a decrease in mRNA and protein expression of ADAM9 in HepG2 cells,which inhibits the malignant phenotype of the cells;miR-1270 targeted the regulation of ADAM9 expression.Conclusion:Lico A may upregulate miR-1270,reduce ADAM9 expression,inhibit proliferation,migration,and invasion of HCC cells,and induce cell apoptosis.