摘要
目的观察长链非编码RNA(LncRNA)LINC00641(LINC00641)在宫颈癌细胞中的表达特征,探讨LINC00641通过微小RNA-181 d-5p(miR-181 d-5p)/真核细胞翻译起始因子4A2(EIF4A2)对宫颈癌细胞增殖的调节作用。方法选择宫颈癌Hela细胞株,建立空白对照组,构建转染pc-DNA3.1的无关序列组､转染pc-DNA3.1-LINC00641的pc-DNA-LINC00641组､转染pc-DNA3.1-LINC00641-siRNA的si-LINC00641组,转染miR-181 d-5p inhibitor的miR-181 d-5p inhibitor组､转染miR-181 d-5p mimic的miR-181 d-5p mimic组､转染siRNA-EIF4A2的si-EIF4A2组。采用CCK-8实验检测细胞增殖活性,采用双荧光素酶报告基因实验观察LINC00641和miR-181 d-5p､miR-181 d-5p､EIF4A2的靶向关系。选取2019年12月至2021年1月陕西中医药大学第二附属医院诊治的45例宫颈鳞癌且未行术前新辅助放､化疗的女性患者作为研究对象,留取其术后肿瘤组织(宫颈鳞癌组织)。另选取45例因子宫肌瘤行子宫全切术,且经病理确诊为慢性宫颈炎的患者,留取其宫颈组织(慢性宫颈炎组织)。采用实时荧光定量聚合酶链反应(qRT-PCR)法检测LINC00641和miR-181 d-5p的表达,采用免疫组化SP法检测EIF4A2和Ki67的表达,采用Pearson检验分析其相关性。结果生物信息分析显示,LINC00641与miR-181 d-5p､miR-181 d-5p､EIF4A2具有可能的结合位点。与空白对照组和无关序列组比较,pc-DNA-LINC00641组､miR-181 d-5p mimic组､si-EIF4A2组细胞增殖活性下降,si-LINC00641组､miR-181 d-5p inhibitor组细胞增殖活性升高。双荧光素酶报告基因实验显示,LINC00641与miR-181 d-5p､miR-181 d-5p､EIF4A2具有靶向关系。挽救实验显示,沉默EIF4A2可部分逆转沉默LINC00641和沉默miR-181 d-5p对细胞增殖的促进作用。与慢性宫颈炎组织比较,宫颈鳞癌组织中LINC00641､miR-181 d-5p低表达,EIF4A2高表达(P<0.05)。Pearson相关分析显示,LINC00641与miR-181 d-5p､EIF4A2､Ki67呈正相关性,miR-181 d-5p与EIF4A2呈负相关性。结论LINC00
Objective To observe the expression characteristics of long non-coding RNA(LncRNA)LINC00641(LINC00641)in cervical cancer cells,investigate the proliferation regulatory effect of LINC00641 on cervical cancer cells by microRNA-181 d-5p(miR-181 d-5p)/eukaryotic translation initiation factor 4A2(EIF4A2).Methods Hela cell line was selected to establish blank control group,the unrelated sequence group(transfected pc-DNA3.1),the pc-DNA-LINC00641 group(transfected pc-DNA3.1-LINC00641),the siLINC00641 group(transfected pc-DNA3.1-LINC00641-siRNA),the miR-181 d-5p inhibitor group(transfected miR-181 d-5p inhibitor),the miR-181 d-5p mimic group(transfected miR-181 d-5p mimic),the si-EIF4A2 group(transfected siRNA-EIF4A2).The proliferation activity was detected by CCK-8 method.The target relationship between LINC00641 and miR-181 d-5p,miR-181 d-5p and EIF4A2 was observed by double luciferase reporter gene experiment.45 cases of cervical squamous cell carcinoma were selected as observation group,45 cases of chronic cervicitis were selected as control group.LINC00641 and miR-181 d-5p were detected by qRTPCR,EIF4A2 and Ki67 were detected by immunohistochemistry method.Correlation was analyzed by Pearson method.Results Potential binding sites was found between LINC00641 and miR-181 d-5p,miR-181 d-5p and EIF4A2 by bioinformatics analysis.Compared with the control group and the unrelated sequence group,the proliferation activity was lower in pc-DNA-LINC00641 group,miR-181 d-5p mimic group and si-EIF4A2 group,was higher in si-LINC00641 group and miR-181 d-5p inhibitor group.Targeted relationship was found between LINC00641 and miR-181 d-5p,miR-181 d-5p and EIF4A2 by double luciferase reporter gene experiment.Silencing of EIF4A2 could partially reverse cell proliferation effect of silence LINC00641 and miR-181 d-5p by rescue experiment.Compared with chronic cervicitis tissues,LINC00641 and miR-181 d-5p were low expression and EIF4A2 was high expression in cervical squamous cell carcinoma tissues(P<0.05).Pearson correlation analysis s
作者
张杨
王刚
陈侠
张云志
秋彤
高飞
ZHANG Yang;WANG Gang;CHEN Xia;ZHANG Yunzhi;QIU Tong;GAO Fei(Second Depart-ment of Oncology,the Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,Shaanxi,China)
出处
《中国性科学》
2022年第2期117-122,共6页
Chinese Journal of Human Sexuality
