miR-143-3p靶向MCL1调节线粒体氧化磷酸化抑制宫颈癌细胞的增殖和迁移研究OA

Objective This study aimed to investigate the mechanism of miR-143-3p in the proliferation and migration of cervical cancer(CC)cells,particularly its effect via targeting the myeloid cell leukemia 1(MCL1)gene to regulate the mitochondrial oxidative phosphorylation process.Methods The CC cell line HeLa was divided into six groups:NC-mimic group(transfected with negative control NC-mimic),miR-143-3p mimic group(transfected with miR-143-3p mimic),oe-NC group(transfected with negative control oe-NC),oe-MCL1 group(transfected with oe-MCL1),miR-143-3p mimic+oe-NC group(co-transfected with miR-143-3p mimic and oe-NC),and miR-143-3p mimic+oe-MCL1 group(co-transfected with miR-143-3p mimic and oe-MCL1).Quantitative real-time polymerase chain reaction(PCR),western blotting,colony formation assays,and wound healing assays were performed to evaluate the expression levels of miR-143-3p and MCL1,as well as cell proliferation and migration.The activities of mitochondrial respiratory chain complexes Ⅰ-Ⅴ,mitochondrial membrane potential,intracellular adenosine triphosphate(ATP)content,and mitochondrial reactive oxygen species(ROS)levels were measured in each group.Results In CC cell lines,miR-143-3p expression was significantly downregulated(F=432.860,P<0.05),whereas the mRNA and protein expression levels of MCL1 were significantly upregulated(F=503.883 and 430.645,respectively,P<0.05).Compared with the NC-mimic group,the miR-143-3p mimic group showed significantly increased miR-143-3p expression and ROS levels(t=19.227 and 14.658,respectively,P<0.05).The mRNA and protein expression levels of MCL1(t=19.941 and 19.078),the number of cell colonies and wound healing rate(t=12.709 and 9.333),the enzymatic activities of mitochondrial respiratory chain complexes Ⅰ-Ⅴ(t=9.177,6.123,7.607,5.777,and 96.12),as well as mitochondrial membrane potential and ATP levels(t=51.783 and 12.335)were all significantly decreased(P<0.05).Compared with the miR-143-3p mimic group,the miR-143-3p mimic+oe-MCL1 group exhibited significantly increased MCL1 mRNA and protein expression(t=7.703 and 8.908),cell colony number and wound healing rate(t=5.282 and 4.713),enzymatic activities of mitochondrial respiratory chain complexes Ⅰ-Ⅴ(t=5.918,4.772,5.637,4.189,and 5.717),as well as mitochondrial membrane potential and ATP levels(t=49.843 and 7.005)(P<0.05).The ROS level was significantly decreased(t=5.648,P<0.05).Conclusion miR-143 is significantly downregulated in CC cell lines.Upregulation of miR-143-3p markedly inhibits oxidative phosphorylation in CC cells,thereby suppressing their proliferation and migration.This effect may be mediated through the targeted inhibition of the downstream gene MCL1 by miR-143-3p.