Effects of miR-30c-5p on mitophagy,mesenchymal transition,and PEG10 levels in human trophoblast cells
Objective To investigate the effects of miR-30c-5p on mitophagy,mesenchymal transition,and PEG10 levels in human trophoblast cells.Methods Human trophoblast cell line HTR-8/SVneo was divided in-to the following groups:blank group(HTR-8/SVneo cells cultured normally),hypoxia group(HTR-8/SVneo cells cultured under hypoxic conditions),miR-30c-5p mimics group(HTR-8/SVneo cells cultured under hypoxic condi-tions transfected with miR-30c-5p mimics),NC-mimics group(HTR-8/SVneo cells cultured under hypoxic condi-tions transfected with NC-mimics),miR-30c-5p inhibitor group(HTR-8/SVneo cells cultured under hypoxic condi-tions transfected with miR-30c-5p inhibitor),and NC-inhibitor group(HTR-8/SVneo cells cultured under hypoxic conditions transfected with NC-inhibitor).qRT-PCR was used to detect the expression of miR-30c-5p in each group of cells.Transwell chamber was used to measure cell invasion.Scratch test kit was used to measure the cell scratch healing rate.ROS level was measured using relevant assay kits.Immunostaining was used to measure the protein lev-els of PEG10,Parkin,and PINK1.Results The expression of miR-30c-5p in the blank group,hypoxia group,miR-30c-5p mimics group,NC-mimics group,miR-30c-5p inhibitor group,and NC-inhibitor group were 1.00±0.00,1.00±0.00,1.61±0.15,1.03±0.13,0.75±0.08,and 0.96±0.10,respectively,showing significant differences(F=45.750,P<0.05).Compared with the blank group,the number of invasive cells,scratch healing rate,PEG10,Par-kin,and PINK1 levels were decreased(P<0.05),while ROS level was increased(P<0.05)in the hypoxia group.There was no significant difference in these parameters between the hypoxia group and the NC-mimics group or the NC-inhibitor group(P>0.05).Compared with the hypoxia group,the number of invasive cells,scratch healing rate,PEG10,Parkin,and PINK1 levels were decreased(P<0.05),while ROS level was increased(P<0.05)in the miR-30c-5p mimics group.Conversely,the number of invasive cells,scratch healing rate,PEG10,Parkin,and PINK1 levels were increased(P<0.05),while ROS level was decreased(P<0.05)in the miR-30c-5p inhibitor group.Con-clusion Inhibition of miR-30c-5p can promote invasion and migration of hypoxic human trophoblasts and reduce ROS expression by promoting mitochondrial autophagy,and this mechanism is associated with the inhibition of PEG10 level.
Human trophoblast cellsMitochondrial autophagymiR-30c-5pMesenchymal transi-tionPaternally expressed gene 10