miR-150靶向调控FOXO4促进宫颈癌C-33A细胞生长与生存

microRNA-150 promotes cervical cancer cell C-33A growth and survival by targeting FOXO4

目的检测miR-150在宫颈癌细胞C-33A中的表达,并探讨其是否通过调控靶向基因FOXO4促进宫颈癌细胞的增殖与凋亡。方法将携带miR-150 mimic(模拟物)和miR-150 inhibitor(抑制物)、阴性对照siRNA经LipofectamineTM2000转染至C-33A细胞中,将细胞分为NC组(阴性对照组)、miR-150模拟物组(转染miR-150 mimic细胞组)及miR-150-in抑制物组(转染miR-150 inhibitor细胞组)。采用流式细胞仪、TUNEL检测法、荧光定量PCR、Western bolt分别检测miR-150对宫颈癌C-33A细胞周期、凋亡以及周期和凋亡相关基因的影响。3'-端非翻译区(UTR)荧光素酶活性分析证实miR-150的直接靶向作用。结果miR-150模拟物促进宫颈癌细胞C-33A的增殖与凋亡,抑制物作用相反,miR-150模拟物促进细胞由G1/G0期进展到S期,从而促进宫颈癌细胞增殖,抑制物相反,miR-150调控几个细胞周期、凋亡相关基因CyclinD1、p27、BIM和FASL的表达;miR-150通过靶向结合FOXO4的3'UTR区从而抑制FOXO4的表达。结论miR-150靶向作用于FOXO4从而调节多种基因的表达以致宫颈癌细胞C-33A的增殖与凋亡。

Objective To detect the expression of miR-150 in cervical cancer cells C-33A and investigate whether it promotes the proliferation and invasion of cervical cancer cells by regulating the target gene FOXO4.Methods The effects of miR-150 on cell cycle and apoptosis,as well as the expression of cycle-and apoptosis-related genes,were determined using flow cytometry,TUNEL assay,qRT-PCR,and Western blot,respectively.The direct target of miR-150 was confirmed using 3' untranslated region (UTR) luciferase reporter assay.Results miR-150 promotes cervical cancer cell C-33A survival and growth,while the inhibition of miR-150 suppresses these actions.miR-150 also induced the cell cycle progressionfrom G1/G0 to S phase,resulting in an enhancement of growth.Several cell cycle-and apoptosis-related genes,CyclinD1,p27,BIM,and FASL were modulated by miR-150.Moreover,miR-150 directly reduced the expression of FOXO4,which regulates the expression of CyclinD1,p27,BIM,and FASL,by targeting its 3' UTR.Conclusion Our data demonstrate that elevated miR-150 targets FOXO4 expression and therefore regulates multiple genes expression,resulting in cervical cancer cell C-33A growth and survival.