LncRNA CCAT1靶向miR-218-5p影响人卵巢癌细胞株SKOV3增殖、侵袭与迁移的作用探讨  

Effects of LncRNA CCAT1 targeting miR-218-5p on the proliferation,invasion and migration of human ovarian cancer cell line SKOV3

作　　者：宋玉芳 张前 刘英杰[1] 刘芳[1] SONG Yufang;ZHANG Qian;LIU Yingjie;LIU Fang(Department of Gynecology,Tangshan Maternal and Child Health Care Hospital,Tangshan,Hebei,063000,China)

机构地区：[1]唐山市妇幼保健院妇科,河北唐山063000

出　　处：《中国优生与遗传杂志》2023年第3期525-532,共8页Chinese Journal of Birth Health & Heredity

基　　金：河北省医学科学研究课题计划(20201482)。

摘　　要：目的探讨长链非编码核糖核酸(LncRNA)结肠癌相关转录因子1(CCAT1)靶向微小核糖核酸-218-5p(miR-218-5p)影响人卵巢癌细胞株SKOV3增殖、侵袭与迁移的作用。方法取人卵巢癌细胞株SKOV3培养,采用脂质体转染法分别将si-CCAT1、pcDNA-CCAT1、CCAT1-NC、miR-218-5p inhibitor、miR-218-5p mimic、miR-NC转染至上述细胞,记为CCAT1下调组、CCAT1上调组、CCAT1对照组、miR-218-5p下调组、miR-218-5p上调组、miR对照组,另取未转染细胞设为空白组,每组6个复孔。细胞计数试剂盒8(CCK-8)法检测细胞增殖;Transwell试验检测细胞侵袭和迁移;反转录定量聚合酶链反应(RT-qPCR)检测细胞中CCAT1、miR-218-5p表达及c-myc、E-钙黏素(E-cadherin)、基质金蛋白酶9(MMP-9)信使核糖核酸(mRNA)表达;蛋白质免疫印迹法(WB)检测细胞中c-myc、E-cadherin和MMP-9蛋白表达。双荧光素酶报告基因实验验证LncRNA CCAT1与miR-218-5p调控关系。结果与空白组和CCAT1对照组比,CCAT1下调组细胞增殖、侵袭和迁移能力降低,CCAT1上调组细胞增殖、侵袭和迁移能力升高(P<0.05);与空白组和miR对照组比,miR-218-5p下调组细胞增殖、侵袭和迁移能力升高,miR-218-5p上调组细胞增殖、侵袭和迁移能力降低(P<0.05)。与空白组和CCAT1对照组比,CCAT1下调组CCAT1表达、c-myc、MMP-9 mRNA及蛋白表达降低,miR-218-5p表达和E-cadherin mRNA及蛋白表达升高,CCAT1上调组CCAT1表达、c-myc、MMP-9mRNA及蛋白表达升高,miR-218-5p表达和E-cadherin mRNA及蛋白表达降低(P<0.05);与空白组和miR对照组比,miR-218-5p下调组CCAT1表达、c-myc、MMP-9 mRNA及蛋白表达升高,miR-218-5p表达和E-cadherin mRNA及蛋白表达降低,miR-218-5p上调组CCAT1表达、c-myc、MMP-9 mRNA及蛋白表达降低,miR-218-5p表达和E-cadherin mRNA及蛋白表达升高(P<0.05)。双荧光素酶报告基因实验结果证实CCAT1靶向调控miR-218-5p。结论CCAT1可提高人卵巢癌细胞株SKOV3的增殖、侵袭与迁移能力,促�Objective To investigate the effect of long non-coding ribonucleic acid(LncRNA)colon cancer associated transcription factor 1(CCAT1)targeting microRNA-218-5p(miR-218-5p)on the proliferation,invasion and migration of human ovarian cancer cell line SKOV3.Methods The human ovarian cancer cell line SKOV3 was cultured,and si-CCAT1,pcDNA-CCAT1,CCAT1-NC,miR-218-5p inhibitor,miR-218-5p mimic,and miR-NC were transfected into the above cells by lipofection method,respectively,which were recorded as CCAT1 down-regulated group,CCAT1 up-regulated group,CCAT1control group,miR-218-5p down-regulated group,miR-218-5p up-regulated group,and miR control group,and the untransfected cells were taken as blank group,with 6 replicate wells in each group.Cell proliferation was detected by cell counting kit 8(CCK-8).Transwell assay was used to detect cell invasion and migration.Reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of CCAT1 and miR-218-5p,and the mRNA expression of c-myc,E-cadherin,matrix metalloprotein 9(MMP-9).The protein expressions of c-myc,E-cadherin and MMP-9 in cells were detected by western blotting(WB).Dual-luciferase reporter gene assay was used to verify the regulatory relationship between LncRNA CCAT1 and miR-218-5p.Results Compared with the blank group and the CCAT1 control group,the cell proliferation,invasion and migration abilities of CCAT1 down-regulated group were decreased,while the cell proliferation,invasion and migration abilities of CCAT1 up-regulated group were increased(P<0.05).Compared with the blank group and the miR control group,the proliferation,invasion and migration abilities of cells in the miR-218-5p down-regulated group were increased,while the proliferation,invasion and migration abilities of cells in the miR-218-5p up-regulated group were decreased(P<0.05).Compared with the blank group and CCAT1 control group,CCAT1 expression,c-myc,MMP-9 mRNA and protein expression in CCAT1 down-regulated group were decreased,while the expression of miR-2

关 键 词：LncRNA CCAT1 miR-218-5p 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]