摘要
目的探讨长链非编码RNA(lncRNA)同源盒D基因簇反义生长相关长非编码RNA(HAGLR)对前列腺癌DU145细胞增殖、侵袭和凋亡的影响及其机制。方法采用实时荧光定量聚合酶链反应(PCR)检测人前列腺癌DU145细胞和正常前列腺上皮RWPE-1细胞中HAGLR的表达水平。将体外培养的DU145细胞分为对照组(未转染)、siNC组(转染阴性对照)和siHAGLR组(转染靶向HAGLR的小干扰RNA),采用实时荧光定量PCR检测各组细胞中HAGLR和微小RNA(miR)-6516-5p的表达水平,噻唑蓝(MTT)法、Transwell小室实验和流式细胞仪检测各组细胞增殖、侵袭和凋亡。采用双荧光素酶报告基因实验检测HAGLR和miR-6516-5p的靶向结合关系。将miR-6516-5p抑制剂(anti-miR-6516-5p)、miRNA抑制剂阴性对照(anti-miR-NC)分别与HAGLR-siRNA共转染后观察下调miR-6516-5p表达对HAGLR低表达的DU145细胞增殖、侵袭和凋亡的影响。多组间比较使用单因素方差分析和LSD-t检验,两组间比较采用独立样本t检验。结果DU145细胞中HAGLR的表达水平(4.38±1.15比1.00±0.06,t=8.727,P<0.05)明显高于RWPE-1细胞,差异有统计学意义。siNC组细胞吸光度值(1.01±0.08比0.98±0.06,t=0.900,P>0.05)、穿膜细胞数[(119.75±8.86)个比(124.56±11.35)个,t=1.002,P>0.05]和细胞凋亡率[(6.62±1.06)%比(6.85±1.12)%,t=0.447,P>0.05]与对照组比较差异均无统计学意义。siHAGLR组细胞吸光度值(0.73±0.05比0.98±0.06,1.01±0.08,t_(对照)=9.603,t_(siNC)=8.904,P<0.05)、穿膜细胞数[(55.64±4.28)个比(124.56±11.35)个,(119.75±8.86)个,t_(对照)=17.043,t_(siNC)=19.543,P<0.05]明显低于对照组、siNC组,而细胞凋亡率[(19.92±3.25)%比(6.85±1.12)%,(6.62±1.06)%,t_(对照)=11.406;t_(siNC)=11.672,P<0.05]均明显高于对照组、siNC组,差异均有统计学意义。双荧光素酶报告基因实验证实miR-6516-5p可与HAGLR靶向结合降低细胞的荧光素酶活性(0.32±0.02比1.00±0.06,t=32.255,P<0.05),差异有统计学意义。siHAGLR+anti-miR-6516-
Objective To investigate the effect of long-chain noncoding RNA(lncRNA)antisense growth related long noncoding RNA homeobox D gene cluster(HAGLR)on the proliferation,invasion and apoptosis of prostate cancer cell line DU145 and its mechanism.Methods The expression of HAGLR in DU145 cells and RWPE-1 cells was detected by real-time fluorescence quantitative PCR.DU145 cells cultured in vitro were divided into control group(not transfected),siNC group(transfected negative control)and siHAGLR group(transfected small interfering RNA targeting HAGLR).The expression levels of HAGLR and microRNA(miR)-6516-5p were detected by real-time fluorescent quantitative PCR,and cell proliferation,invasion and apoptosis were tested by methyl thiazolyl tetrazolium(MTT)method,Transwell chamber test and flow cytometry.The target binding relationship between HAGLR and miR-6516-5p was detected by double luciferase reporter gene assay.After co-transfection of miR-6516-5p inhibitor(anti-miR-6516-5p)and miRNA inhibitor negative control(anti-miR-NC)with HAGLR-siRNA,respectively,the effect of miR-6516-5p down-regulation on the proliferation,invasion and apoptosis of DU145 cells with low expression of HAGLR was observed.One-way analysis of variance and LSD-t test were used for comparison between multiple groups,and independent sample t test was used for comparison between two groups.Results Compared with RWPE-1 cells,the expression level of HAGLR(4.38±1.15 vs.1.00±0.06,t=8.727,P<0.05)in DU145 cells was significantly increased.There were no significant differences in cell absorbance value(1.01±0.08 vs.0.98±0.06,t=0.900,P>0.05),number of transmembrane cells(119.75±8.86 vs.124.56±11.35,t=1.002,P>0.05)and apoptosis rate[(6.62±1.06)%vs.(6.85±1.12)%,t=0.447,P>0.05]between siNC group and control group.The cell absorbance value(0.73±0.05 vs.0.98±0.06,1.01±0.08,t_(control)=9.603,t_(siNC)=8.904,P<0.05)and number of transmembrane cells(55.64±4.28 vs.124.56±11.35,119.75±8.86,t_(control)=17.043,t_(siNC)=19.543,P<0.05)in siHAGLR group were sign
作者
田亚明
赵辉
王淑娟
张大
武新超
崔西春
杨帆
Tian Yaming;Zhao Hui;Wang Shujuan;Zhang Da;Wu Xinchao;Cui Xichun;Yang Fan(Department of Pediatric Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Hospital Infection Control,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第4期703-707,共5页
Chinese Journal of Experimental Surgery
