长链非编码RNA PCED1B-AS1靶向miR-485-5p抑制宫颈癌细胞增殖、迁移和侵袭的实验研究  

LncRNA PCED1B-AS1 knockdown inhibits proliferation,migration and invasion of cervical cancer via targeting miR-485-5p

作　　者：刘明博[1] 董青青 周博[1] 刘东斌 王跃伟[1] 吴广银[1] LIU Mingbo;DONG Qingqing;ZHOU Bo;LIU Dongbin;WANG Yuewei;WU Guangyin(Department of Cancer Center, Henan Provincial People's Hospital, Zhengzhou 450000, China)

机构地区：[1]河南省人民医院肿瘤中心,郑州450000

出　　处：《临床肿瘤学杂志》2022年第5期385-392,共8页Chinese Clinical Oncology

摘　　要：目的 探讨PCED1B-AS1在宫颈癌中的表达及其对预后、细胞增殖、迁移和侵袭能力的影响。方法 选取2013年1月至2016年12月河南省人民医院80例宫颈癌患者标本。采用荧光定量PCR(qRT-PCR)检测PCED1B-AS1及靶基因miR-485-5p的表达。采用Kaplan-Meier法和Cox比例风险回归模型分析PCED1B-AS1表达对预后的影响。将PCED1B-AS1低表达质粒(si-PCED1B-AS1)及其阴性对照质粒(si-NC)、miR-485-5p过表达质粒(miR-485-5p模拟物)及其阴性对照质粒(NC模拟物)、miR-485-5p低表达质粒(miR-485-5p抑制物)及其阴性对照质粒(NC抑制物)分别转染至宫颈癌HeLa细胞。采用CCK-8法和Transwell实验检测宫颈癌细胞增殖、迁移及侵袭能力的变化。通过生物信息学预测PCED1B-AS1的靶基因。采用双荧光素酶报告基因检测、RNA免疫共沉淀(RIP)实验及挽救(Rescue)实验验证PCED1B-AS1和miR-485-5p的靶向调控关系。结果 PCED1B-AS1在宫颈癌组织中高表达,且高表达患者有着较差的总体生存期(OS)。CCK-8检测结果显示si-PCED1B-AS1组细胞的增殖能力显著低于si-NC组(P<0.01)。Transwell实验结果显示si-PCED1B-AS1组的迁移细胞数为223±15,显著低于si-NC组的463±15,差异有统计学意义(P<0.001)。同时,si-PCED1B-AS1组的侵袭细胞数为90±10,显著低于si-NC组的400±10,差异有统计学意义(P<0.001)。PCED1B-AS1表达与miR-485-5p表达呈负相关(r=-0.823,P<0.001)。双荧光素酶报告基因与RIP实验证实了PCED1B-AS1与miR-485-5p之间的相互作用。挽救实验表明,miR-485-5p低表达可以抵消si-PCED1B-AS1对细胞增殖、迁移和侵袭能力的抑制作用(P<0.001)。结论 PCED1B-AS1在宫颈癌中表达上调,并且下调PCED1B-AS1可通过靶向负调控miR-485-5p抑制宫颈癌细胞的增殖、迁移和侵袭能力。Objective To investigate the expression level of PCED1B-AS1 in cervical cancer,and its relationship with the prognosis of patients,and proliferation,migration,invasion of human cervical cancer HeLa cells.Methods A total of 80 cervical cancer patients from the Henan Provincial People's Hospital between January 2013 and December 2016 were enrolled.The expression levels of PCED1B-AS1 and miR-485-5p were evaluated using real-time quantitative PCR(qRT-PCR).Survival analysis was performed using Kaplan-Meier curves and Cox proportional hazards regression model.PCED1B-AS1 knockdown plasmid(si-PCED1B-AS1)and its negative control(si-NC),miR-485-5p over-expression plasmid(miR-485-5p mimics)and its negative control(NC mimics),as well as miR-485-5p knockdown plasmid(miR-485-5p inhibitor)and its negative control(NC inhibitor)were transfected into HeLa cells,respectively.The effects of PCED1B-AS1 on cell proliferation,migration and invasion capacities were analyzed using CCK-8 and Transwell assays.Target genes of PCED1B-AS1 were assessed using bioinformatics,luciferase reporter assay,RNA immunocoprecipitation(RIP)assay and rescue experiments.Results PCED1B-AS1 was upregulated in human cervical cancer tissues,and its high expression was associated with poor prognosis of patients.CCK-8 assays demonstrated that the proliferation activity of HeLa cells in si-PCED1B-AS1 group was significantly lower than that in si-NC group(P<0.01).Transwell assays revealed that the number of migrating cells was 223±15,which was significantly lower than that in si-NC group(463±15),and the difference was statistically significant(P<0.001).Meanwhile,the number of invading cells was 90±10,which was significantly lower than that in si-NC group(400±10),and the difference was statistically significant(P<0.001).PCED1B-AS1 expression was negatively correlated with miR-485-5p expression(r=-0.823,P<0.001).Luciferase reporter assays and RIP confirmed that miR-485-5p was a direct target of PCED1B-AS1.Rescue assays demonstrated that miR-485-5p downregulation

关 键 词：宫颈癌 PCED1B-AS1 miR-485-5p 促癌基因 

分 类 号：R737.33[医药卫生—肿瘤]