miR-186下调KIF3C抑制PI3K/AKT信号通路抗肺癌细胞的增殖、迁移和侵袭  

MicroRNA-186 inhibits the proliferation,migration and invasion of lung cancer cells by inhibiting KIF3C and PI3K/AKT signaling pathway

作　　者：刘海旺[1] 刘燃[2] 郝美玲[1] 赵醒[1] 李春辉[1] Haiwang Liu;Ran Liu;Meiling Hao;Zhao Xing;Chunhui Li(Affiliated Hospital of Chengde Medical College Department of Pathology,Chengde 067000;Affiliated Hospital of Chengde Medical College Department of anesthesia,Chengde 067000)

机构地区：[1]承德医学院附属医院病理科,承德067000 [2]承德医学院附属医院麻醉科,承德067000

出　　处：《湖南师范大学学报（医学版）》2022年第5期21-28,共8页Journal of Hunan Normal University(Medical Sciences)

基　　金：2021年度河北省医学科学研究课题计划项目(20211020)。

摘　　要：目的 :探究驱动蛋白家族成员3C(kinesin family member 3C,KIF3C)和微小RNA(mircoRNA,miR)-186对人肺癌细胞增殖、迁移和侵袭的影响及其调控机制。方法 :体外培养人肺癌A549细胞系,然后转染miR-186mimics、miR-186 inhibitor、siRNA(si)-KIF3C、miR-186 inhibitor+si-KIF3C及mimic NC、inhibitor NC和si-NC至A549细胞,分别设为miR-186 mimic组、miR-186 inhibitor组、si-KIF3C组、miR-186 inhibitor+si-KIF3C组及NC组(mimic NC组、inhibitor NC组和si-NC组),非转染的细胞设为Con组。用实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)测定A549细胞中miR-186及KIF3C基因表达水平;蛋白免疫印迹(western blotting,WB)法测定KIF3C和磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶(phosphatidylinositol 3 kinase/serine threonine protein kinase,PI3K/AKT)通路相关蛋白表达水平;活细胞计数试剂盒-8(cell counting kit-8,CCK-8)测定细胞活力;5-乙炔基-2’脱氧尿嘧啶核苷(5-ethynyl-2’deoxyuracil nucleoside,EdU)法测定细胞增殖;Transwell小室测定细胞的迁移和侵袭。结果 :miR-186 mimic组miR-186表达水平显著高于mimic NC组(P<0.05),miR-186 inhibitor组miR-186表达水平显著低于inhibitor NC组(P<0.05),si-KIF3C组KIF3C mRNA和蛋白表达水平显著低于si-NC组(P<0.05),以上实验证明了miR-186 mimic、miR-186 inhibitor和si-KIF3C转染成功。与NC组相比,miR-186 mimic组和siKIF3C组细胞活力、增殖率、迁移数、侵袭数及KIF3C、p-PI3K和p-AKT蛋白表达水平显著降低,miR-186 inhibitor组则显著升高(P<0.05);miR-186 inhibitor+si-KIF3C组上述指标显著低于miR-186 inhibitor组,显著高于siKIF3C组(P<0.05)。结论 :过表达miR-186通过下调KIF3C抑制肺癌细胞的增殖、迁移和侵袭,其作用机制可能是抑制PI3K/AKT信号通路有关。Objective To investigate the role of kinesin family member 3C(KIF3C)and microRNA-186(miR-186on the proliferation,migration and invasion of human non-small cell lung cancer cells. Methods Human lung cancer A549cell line was cultured in vitro and divided into miR-186 mimic group,miR-186 inhibitor group,si-KIF3C group,miR-186inhibitor+si-KIF3C group and NC group(mimic NC group,inhibitor NC group and si-NC group. Dysregulation of miR-186 and KIF3C was obtained using cell transfection,and transfection efficiency was determined by real-time quantitative PCR(RT-qPCR)and western blot(WB). The activation of phosphatidylinositol-3 kinase/serine-threonine protein kinase(PI3K/AKT)pathway was detected by WB. Results miR-186 expression was highly increased and significantly decreased with miR-186 mimic or inhibition transfection;KIF3C mRNA and protein expression levels were significantly decreased by transfecting KIF3C small interfering RNA. Moreover,cell viability,proliferation,migration,invasion and protein levels of phosphorylated(p)-PI3K and p-AKT were overall inhibited by miR-186 upregulation and KIF3C downregulation,and were uniformly enhanced by silencing miR-186 and promoting KIF3C. Notably,miR-186 inhibition-induced promotions were attenuated by KIF3C downregulation,and KIF3C inhibition-induced suppressions were counteracted by miR-186 downregulation as well.Conclusion miR-186 might downregulate KIF3C to suppress the proliferation,migration and invasion of A549 cells through inhibiting PI3K/AKT signaling pathway,indicating a possible miR-186/KIF3C regulation in lung cancer.

关 键 词：肺癌 miR-186 驱动蛋白家族成员3C 磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶信号通路 增殖 迁移 侵袭 

分 类 号：R363[医药卫生—病理学]