微小RNA-600在卵巢癌组织的表达及其靶向调控对氧磷酶3对卵巢癌细胞株OVCAR-3侵袭和迁移的影响  

Expression of microRNA-600 in ovarian cancer tissues and its targeted regulation effect of paraoxonase 3 on invasion and migration of ovarian cancer cell line OVCAR-3

作　　者：李航 杨蝶 刘慧芝 LI Hang;YANG Die;LIU Hui-Zhi(Department of Medical Services,Affiliated Hospital,Zunyi Medical University,Zunyi 563003,China)

机构地区：[1]遵义医科大学附属医院医务处,遵义563003 [2]遵义市第一人民医院血管超声诊断室,遵义563002

出　　处：《中国免疫学杂志》2019年第19期2357-2361,2368,共6页Chinese Journal of Immunology

基　　金：贵州省教育厅计划项目[No.KY(2017)045]

摘　　要：目的:探讨微小RNA(miRNA)-600在卵巢癌组织及细胞系的表达及其靶向调控对氧磷酶3对卵巢癌细胞株OVCAR-3侵袭和迁移的机制。方法:收集2017年1月至2018年10月我院妇产科经手术治疗的卵巢癌患者共50例。使用Lipofectamine 2000将miRNA-600模拟物、miRNA-600抑制剂、模拟物与抑制剂对照物、对氧磷酶3(PON3)及其对照载体等转染到卵巢癌细胞。实时荧光定量聚合酶链反应(qRT-PCR)检测miRNA-600和PON3基因相对表达量。蛋白印迹检测法(WB)检测PON3及侵袭和迁移相关分子E-钙黏素、N-钙黏素、MMP-2、MMP-9的蛋白分子水平。应用划痕试验和Transwell实验分别测定细胞迁移和侵袭能力。免疫组织化学法检测组织中PON3、E-钙黏素、N-钙黏素、MMP-2、MMP-9的水平。结果:与癌旁组织(6.11±1.15)相比,癌组织(3.81±0.69)中miRNA-600的相对表达量显著降低(t=7.139,P<0.05)。与模拟物对照[miRNA-600:2.13±0.18;N-钙黏素:4.78±0.69;E-钙黏素:2.07±0.24;MMP-2:5.16±0.73;MMP-9:4.44±0.62;迁移率:(73.45±5.18)%;细胞侵袭数量:47.22±6.90]相比,miRNA-600模拟物[miRNA-600:7.88±0.69;N-钙黏素:2.21±0.45;E-钙黏素:4.67±0.20;MMP-2:2.04±0.12;MMP-9:2.39±0.20;迁移率:(37.63±3.76)%;细胞侵袭数量:19.34±4.63]的miRNA-600与E-钙黏素显著增高,而N-钙黏素、MMP-2、MMP-9、迁移率和细胞侵袭数量显著降低(P<0.05);与抑制剂对照[miRNA-600:2.15±0.19;N-钙黏素:1.89±0.22;E-钙黏素:4.92±0.42;MMP-2:1.76±0.09;MMP-9:1.88±0.12;迁移率:(41.45±3.97)%;细胞侵袭数量:41.56±7.98]相比,miRNA-600抑制剂[miRNA-600:0.26±0.08;N-钙黏素:4.67±0.60;E-钙黏素:1.85±0.07;MMP-2:4.38±0.57;MMP-9:4.55±0.60;迁移率:(84.12±6.50)%;细胞侵袭数量:62.89±9.57]的miRNA-600与E-钙黏素显著降低,而N-钙黏素、MMP-2、MMP-9、迁移率和细胞侵袭数量显著增高(P<0.05)。与模拟物对照[迁移率:(46.45±3.88)%;细胞侵袭数量:49.76±4.26]相比,miRNA-600模拟物[迁移率:(22.09±1.68)%;细胞侵袭�Objective:To investigate the expression of microRNA(miRNA)-600 in ovarian cancer tissues and cell lines and to explore the mechanism of paraoxonase 3 invasion and migration of ovarian cancer cell line OVCAR-3.Methods:A total of 50 patients with ovarian cancer who underwent surgery in our department from January 2017 to October 2018 were enrolled.miRNA-600 mimics,miRNA-600 inhibitor,mimics and inhibitor control,paraoxonase 3(PON3)and its control vector were transfected into ovarian cancer cells by Lipofectamine 2000.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the relative expression of miRNA-600 and PON3 genes.Western blot was used to detect the protein levels of PON3 and invasion and migration related molecules E-cadherin,N-cadherin,MMP-2 and MMP-9.The cell migration and invasion ability were determined by the scratch test and the Transwell test,respectively.The levels of PON3,E-cadherin,N-cadherin,MMP-2,MMP-9 in tissues were detected by immunohistochemistry.Results:Compared with adjacent tissues(6.11±1.15),the relative expression of miRNA-600 in cancer tissues(3.81±0.69)was significantly decreased(t=7.139,P<0.05).Compared with the mimics control[miRNA-600:2.13±0.18;N-cadherin:4.78±0.69;E-cadherin:2.07±0.24;MMP-2:5.16±0.73;MMP-9:4.44±0.62;mobility:(73.45±5.18)%;cell invasion number:47.22±6.90),miRNA-600 mimics(miRNA-600:7.88±0.69;N-cadherin:2.21±0.45;E-cadherin:4.67±0.20;MMP-2:2.04±0.12;MMP-9:2.39±0.20;mobility:(37.63±3.76)%;cell invasion number:19.34±4.63]showed significantly increased miRNA-600 and E-cadherin,while N-cadherin,MMP-2,MMP-9,mobility and cell invasion were significantly reduced(P<0.05).Compared with the inhibitor control[miRNA-600:2.15±0.19;N-cadherin:1.89±0.22;E-cadherin:4.92±0.42;MMP-2:1.76±0.09;MMP-9:1.88±0.12;migration:(41.45±3.97)%;cell invasion number:41.56±7.98],miRNA-600 inhibitors[miRNA-600:0.26±0.08;N-cadherin:4.67±0.60;E-cadherin:1.85±0.07;MMP-2:4.38±0.57;MMP-9:4.55±0.60;mobility:(84.12±6.50%);cell invasion number:62.89±9.57]s

关 键 词：卵巢癌 微小RNA 迁移 侵袭 对氧磷酶3 

分 类 号：R737.31[医药卫生—肿瘤]