Influences of circEIF6 on the proliferation,apoptosis and gemcitabine sensitivity of pancreatic cancer cells by regulating the miR-129-5p/TRAF6 axis
Objective:To investigate the influences of circular RNA eukaryotic initiation factor 6(circEIF6)on the proliferation,apoptosis and gemcitabine(GEM)sensitivity of pancreatic cancer(PC)cells by regulating miR-129-5p/tumor necrosis factor receptor associated factor 6(TRAF6)axis.Methods:PC cells SW1990 were divided into a control group(normal culture),a si-NC group(transfected with si-NC),a si-circEIF6 group(transfected with si-circEIF6),a si-circEIF6+inhibitor NC group(co-transfected with si-circEIF6 and inhibitor NC),and a si-circEIF6+miR-129-5p inhibitor group(co-transfected with si-circEIF6 and miR-129-5p inhibitor).The expression levels of circEIF6 and miR-129-5p were detected by RT-qPCR.The proliferation ability of cells was detected by MTT assay.Cell apoptosis was detected by flow cytometry.Western blotting was applied to detect the expression of TRAF6,proliferating cell nuclear antigen(PCNA),Bax and caspase-3.MTT assay was used to detect the effect of different concentrations of GEM on the proliferation inhibition rate.Double luciferase reporter gene experiment was used to verify the relationship between circEIF6 and miR-129-5p,between miR-129-5p and TRAF6 respectively.Results:Compared with the control group and si-NC group,the expression of circEIF6,OD490 value,the protein expression of TRAF6 and PCNA in si-circEIF6 group were decreased,the expression of miR-129-5p,apoptosis rate,the protein expression of Bax and caspase-3 were increased.Compared with the si-circEIF6 group and si-circEIF6+inhibitor NC group,there was no obvious difference in the expression of circEIF6 in the si-circEIF6+miR-129-5p inhibitor group,OD490 value,and the protein expression of TRAF6 and PCNA were increased,the expression of miR-129-5p,apoptosis rate,the protein expression of Bax and caspase-3 were decreased.The proliferation inhibition rate was obviously increased in a dose-dependent manner under GEM treatment.Compared with the control group,the proliferation inhibition rate and apoptosis rate under GEM treatment were obviously higher,knocking-down circEIF6 could increase the proliferation inhibition rate and apoptosis rate under GEM treatment.However,miR-129-5p inhibitor could weaken the above effects of knockdown circEIF6.circEIF6 negatively regulated the expression of miR-129-5p,and miR-129-5p negatively regulated the expression of TRAF6.Conclusion:Knocking down circEIF6 may down-regulate the expression of TRAF6 by targeting miR-129-5p,thereby inhibiting the proliferation of PC cells,promoting apoptosis,and increasing the sensitivity of PC cells to GEM.
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