摘要
目的:长链非编码RNA(long non-codingRNA,LncRNA)是机体重要的转录及转录后调控分子,近年其与肿瘤细胞恶性表型的关系逐步被揭示,本研究旨在探究垂体特异性转录因子-3相关长链非编码RNA(pit-oct-unc class 3 homeobox 3 related longnon-codingRNA,Linc-POU3F3)在食管癌中的表达特征及其与食管癌细胞放射治疗(以下简称放疗)抵抗(radiation resistance,IR)及肿瘤干细胞(cancer stemcells,CSCs)标志物表达的关系。方法:采用生物信息学方法检索公共数据库中Linc-POU3F3在食管癌中的表达特征及潜在的相互作用分子。收集42例食管癌组织样本及相应的癌旁组织,培养人正常食管上皮细胞HEEC及人食管癌细胞ECA109、TE-1、TE-2、TE-13。采用实时荧光定量PCR(qPCR)技术检测食管癌组织及细胞中Linc-POU3F3的表达水平。以梯度递增剂量的射线诱导食管癌细胞系TE-13 IR的形成作为IR组细胞,同等条件以0 Gy剂量处理TE-13细胞系作为对照组(Control)细胞;同时采用细胞转染技术,以实验组细胞为模型构建随机干扰序列组(siControl)细胞及靶向Linc-POU3F3的干扰组(siLinc-POU3F3)细胞,在ECA109细胞中转染空白及过表达Linc-POU3F3载体作为空白组(Vector)及过表达组(oeLinc-POU3F3)。采用qPCR技术及蛋白质印迹法分别检测CSCs标志物CD44、CD133及CD90 mRNA及蛋白质的表达水平;四唑化合物MTS[3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt]检测不同放射剂量下细胞活力;以克隆形成实验验证IR细胞的抵抗能力。结果:生物信息分析结果显示Linc-POU3F3的表达与食管癌患者肿瘤进展及预后不良相关;Linc-POU3F3 mRNA在食管癌组织及细胞系中的表达均显著高于正常癌旁组织及正常细胞(均P<0.01),IR细胞中Linc-POU3F3、CD44、CD133及CD90 mRNA及蛋白质表达均显著高于Control细胞(均P<0.01);siLinc-POU3F3细胞Linc-POU3F3表达显著低于siControl细胞(P<0.01),抑制率
Objective:Long non-coding RNA(LncRNA)is an important transcriptional and posttranscriptional regulatory molecule in the body.In recent years,relationship between LncRNA and malignant phenotype of tumor cells has been revealed gradually.This study aims to investigate the expression characteristics of pit-oct-unc class 3 homeobox 3 related long non-coding RNA(Linc-POU3F3)in esophageal cancer and its relationship with radiation resistance(IR)as well as the expressions of cancer stem cell(CSC)markers in esophageal cancer cells.Methods:The expression characteristics and potential interaction molecules of Linc-POU3F3 in esophageal cancer were collected from the public database via bioinformatics retrieval.Forty-two pair samples of esophageal cancer tissues and corresponding adjacent tissues were collected.Human normal esophageal epithelial cells(HEEC)and human esophageal cancer cell lines(ECA109,TE-1,TE-2,TE-13)were cultured.Real-time quantitative PCR(qPCR)was used to detect the expression level of Linc-POU3F3 in clinical tissues and cells.The formation of TE-13 IR cell line induced by different doses of radiation served as IR group cells,and the same condition treated with 0 Gy dose was set as control group(control)cells.Meanwhile,we used cell transfection technology to construct random interference sequence(siControl)cells and interference(siLinc-POU3F3)cells.In ECA109 cells,we transfected blank and over expressed Linc-POU3F3 plasmids as vector and over-expressed group(oeLinc-POU3F3).The mRNA and protein expressions of CD44,CD133 and CD90 were detected by qPCR and Western blotting,respectively.MTS[3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt]was used to detect the cell viability under different radiation doses,and the resistance of IR cells was verified by clone formation experiment.Results:The expression of Linc-POU3F3 was correlated with the tumor progression and poor prognosis of esophageal cancer.The level of Linc-POU3F3 mRNA expression was significantly hig
作者
陈一川
唐敬群
李乐之
卢婷
CHEN Yichuan;TANG Jingqun;LI Lezhi;LU Ting(Department of Cardiovascular Surgery,Second Xiangya Hospital,Central South University,Changsha 410011;Department of Thoracic Surgery,Second Xiangya Hospital,Central South University,Changsha 410011;Clinic Nursing Teaching and Research Section,Second Xiangya Hospital,Central South University,Changsha 410011,China)
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2021年第6期583-590,共8页
Journal of Central South University ：Medical Science
