miR-34b-3p靶向FDX1抑制小儿神经母细胞瘤细胞生长和凋亡  

miR-34b-3p targeting FDX1 suppressed the growth and apoptosis of neuroblastoma cells in children

作　　者：冯祯祯 赵煜[2] 黄贵祥[2] 路翔宇 王宇 陈丹 Feng Zhenzhen;Zhao Yu;Huang Guixiang;Lu Xiangyu;Wang Yu;Chen Dan(epartment of Pediatric Surgery,Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital,University of Electronic Science&Technology of China,Chengdu 610072,China;Department of Emergency Medicine,Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital,Chengdu 610072,China;Department of Hepatobiliary Surgery,Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital,Chengdu 610072,China;Institute of Disaster&Emergency Medicine,Sichuan Provincial People's Hospital,Chengdu 610072,China)

机构地区：[1]四川省医学科学院·四川省人民医院,电子科技大学附属医院小儿外科,成都610072 [2]四川省医学科学院·四川省人民医院,电子科技大学附属医院急诊科,成都610072 [3]四川省医学科学院·四川省人民医院,电子科技大学附属医院肝胆外科,成都610072 [4]四川省人民医院灾难医学与急诊医学研究所,成都610072

出　　处：《中华小儿外科杂志》2024年第3期262-268,共7页Chinese Journal of Pediatric Surgery

基　　金：成都市医学科研项目(2022708)。

摘　　要：目的探究miR-34b-3p在神经母细胞瘤(neuroblastoma,NB)细胞增殖和凋亡过程中的调节作用。方法体外培养人正常背根神经节细胞和NB细胞系,搜集NB组织和邻近的正常组织。qRT-PCR检测NB组织和细胞中miR-34b-3p和FDX1的表达;CCK-8法、克隆形成实验和流式细胞术检测NB细胞的增殖和凋亡;双荧光素酶报告基因分析FDX1 mRNA 3'非翻译区(3'-untranslated region,3'-UTR)和miR-34b-3p之间的靶点关系;Western blot检测相关蛋白的表达。采用SPSS 23.0和GraphPad Prism 6.01进行Student's t-test检验。结果与对照组比较,miR-34b-3p在NB肿瘤组织和癌细胞系(SK-N-BE、SK-N-SH、SH-SY5Y和LAN-6)中均下调(P<0.05)。与对照组比较,miR-34b-3p过表达抑制了癌细胞SK-N-BE和SH-SY5Y的生长,并诱导凋亡增加(P<0.05)。萤光素酶报告基因证实,miR-34b-3p可以与FDX13'-UTR结合,抑制NB细胞中FDX1的表达。此外,FDX1过表达有效逆转了miR-34b-3p对NB细胞生长和凋亡的抑制作用。与对照组比较,miR-34b-3p稳定转染SH-SY5Y细胞抑制了异种移植瘤的生长和肿瘤重量(P<0.05)。结论miR-34b-3p可能通过靶向FDX1在NB中发挥肿瘤抑制作用,是一种新颖的NB诊断和治疗的生物标志物。Objective To explore the regulatory role of miR-34b-3p in the proliferation and apoptosis of neuroblastoma(NB)cells.Methods Human normal dorsal root ganglion cell and NB cell lines were cultured in vitro,and NB tissues and adjacent normal tissues were harvested.Quantitative real-time polymerase chain reaction(qRT-PCR)was employed for detecting the expressions of miR-34b-3p and FDX1 in NB tissues and cells.The proliferation and apoptosis of NB cells were detected by CCK-8 assay,colony formation assay and flow cytometry.Target relationship between ferriredoxin 1(FDX1)mRNA 3'untranslated region(3'-UTR)and miR-34b-3p was examined by dual-luciferase reporter gene.Western blot was utilized for detecting the expression of related proteins.And SPSS 23.0 and GraphPad Prism 6.01 were utilized for Student's t-test.Results As compared with control group,miR-34b-3p was down-regulated in NB tumor tissues and cell lines(SK-N-BE,SK-N-SH,SH-SY5Y&LAN-6)(P<0.05).An over-expression of miR-34b-3p suppressed the growth of SK-N-BE and SH-SY5Y cells and induced their apoptosis(P<0.05).Luciferase reporter gene confirmed that miR-34b-3p conjugated with FDX13'-UTR and arrested the expression of FDX1 in NB cells.In addition,an over-expression of FDX1 effectively reversed the inhibitory effect of miR-34b-3p on the growth and apoptosis of NB cells.As compared with control group,stable transfection of miR-34b-3p into SH-SY5Y cells suppressed the growth and tumor weight of xenograft tumor(P<0.05).Conclusions miR-34b-3p may play a tumor suppressor role in NB through targeting FDX1.It is a novel biomarker for diagnosing and treating NB.

关 键 词：神经母细胞瘤 微RNA 细胞增殖 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]