Effect of lncRNA-AFAP1-AS1 targeting miR-195-5p on biological behavior of hepatocellular carcinoma cells
LI Junhui
Yeermaike Ahati
GU Peng
Abstract：Objective To investigate the effects of long non-coding RNA(lncRNA)-AFAP1-AS1 and microRNA(miR)-195-5p on the biological behavior of hepatocellular carcinoma(HCC)cells.Methods Human HCC cell lines(HepG2,Hep3B,SMMC-7721,Bel7402,SK-hep1 cells)and normal human liver cells(HL-7702 cells)were routinely cultured.The expression levels of AFAP1-AS1 and miR-195-5p were detected by real-time fluorescence quantitative PCR to select the experimental cells.The experimental cells in logarithmic growth phase were randomly assigned to different groups:si-AFAP1-AS1 group,si-NC group,miR-195-5p mimics group,mimics-NC group,miR-195-5p inhibitor+si-NC group,miR-195-5p inhibitor+si-AFAP1-AS1 group,inhibitor-NC+si-AFAP1-AS1 group,and inhibitor-NC+si-NC group,respectively.Subsequently,the corresponding plasmids were transfected into the cells using Lipofectamine 2000 reagent based on the grouping.Cell proliferation was assessed by CCK-8 and colony formation experiments,while apoptosis and cell cycle were analyzed using flow cytometry.Cell migration and invasion capabilities were evaluated by Transwell experi-ment,and the targeted relationship between AFAP1-AS1 and miR-195-5p was confirmed via dual luciferase reporter gene assay.Results In HCC cells,AFAP1-AS1 expression was higher than that in normal liver cells(P<0.05),while miR-195-5p expression was lower than that of the normal liver cells(P<0.05).Given that SK-hep1 cells exhibited the highest AFAP1-AS1 expression and the lowest miR-195-5p expression,subsequent investigations were carried out using SK-hep1 cells for experimental purposes.Compared with the si-NC group,the si-AFAP1-AS1 group displayed reduced AFAP1-AS1 expression,decreased cell proliferation capacity,reduced migration and invasion cells,a higher cell population in the G0/G1 phase,and an elevated apoptosis rate(all P<0.05).The Starbase v3.0 online database predicted that there were com-plementary binding sites between AFAP1-AS1 and miR-195-5p.Findings from the dual-luciferase reporter gene assay indi-cated that there was significant difference in the relative luciferase activity between the miR-NC and miR-195-5p after co-transfection of WT-AFAP1-AS1(P<0.05),while no significant difference was found after co-transfection of MUT-AFAP1-AS1(P>0.05).Compared with the inhibitor-NC+si-NC group,the si-NC+miR-195-5p inhibitor group demonstrated en-hanced cell proliferation,increased migration and invasion,reduced cell population in the G0/G1 phase,and decreased apoptosis rate(all P<0.05).Conversely,the si-AFAP1-AS1+miR-195-5p inhibitor group exhibited the opposite cellular re-sponses.Conclusion The lncRNA AFAP1-AS1 potentially plays a role in regulating various biological processes in HCC cells,including proliferation,migration,invasion,and apoptosis,through its competitive interaction with miR-195-5p.
Keywords：hepatocellular carcinomalong non-coding RNAAFAP1-AS1miR-195-5pcell biological behavior
Publication Date：2024-07-05
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 40-44 )