长链非编码RNA HOX转录反义RNA靶向miR-206对结直肠癌细胞迁移、侵袭的影响  

Effect of long non-coding RNA HOX transcript antisense inter genic RNA targeting miR-206 on migration and invasion of colorectal cancer cells

作　　者：薛强 武雪亮[2] 孙光源 杨东东[3] 李坤[4] XUE Qiang;WU Xueliang;SUN Guangyuan;YANG Dongdong;LI Kun(Department of General Surgery,the First Hospital of Zhangjiakou,Hebei Province,Zhangjiakou075000,China;Cancer Research Institute,the First Affiliated Hospital of Hebei North University,Hebei Province,Zhangjiakou075000,China;Department of General Surgery,the First Affiliated Hospital of Hebei North University,Hebei Province,Zhangjiakou075000,China;Department of Pathology,the First Affiliated Hospital of Hebei North University,Hebei Province,Zhangjiakou075000,China)

机构地区：[1]张家口市第一医院普通外科,河北省张家口075000 [2]河北北方学院附属第一医院肿瘤研究所,河北省张家口075000 [3]河北北方学院附属第一医院普通外科,河北省张家口075000 [4]河北北方学院附属第一医院病理科,河北省张家口075000

出　　处：《中国医药导报》2023年第34期26-30,共5页China Medical Herald

基　　金：河北省卫生健康委员会医学科学研究课题计划项目(20210954)。

摘　　要：目的探讨长链非编码RNA(lncRNA)HOX转录反义RNA(HOTAIR)靶向miR-206对结直肠癌细胞侵袭和迁移的影响。方法常规培养人结直肠癌HT-29细胞,将合成的HOTAIR si RNA和siRNA control转染于HT-29细胞,设置为si-HOTAIR组、si-con组,未进行转染的HT-29细胞设置为空白组。实时荧光定量聚合酶链反应测定转染效果,Transwell和划痕愈合实验检测HT-29细胞侵袭和迁移情况。生物信息学软件分析HOTAIR与miR-206区域结合位点,构建突变型(HOTAIR 3’UTR mut)和野生型(HOTAIR 3’UTR wt)萤光素酶报告载体,将HOTAIR 3’UTR wt、HOTAIR 3’UTR mut与miR-206 mimic或miR-NC载体共转染至HT-29细胞中,设为HOTAIR 3’UTRwt/miR-NC组、HOTAIR3’UTRwt/miR-206组、HOTAIR3’UTRmut/miR-NC组,HOTAIR3’UTRmut/miR-206组,培养48 h后检测萤光素酶活性。将HOTAIR si RNA和miR-206 inhibitor共转染于HT-29细胞,设置si-HOTAIR+anti-NC组和si-HOTAIR+anti-miR-206组,用Transwell和划痕愈合实验分析细胞侵袭、迁移水平。结果空白组与si-con组HT-29细胞中HOTAIR m RNA的表达水平比较,差异无统计学意义(P>0.05);与si-con组比较,si-HOTAIR组HT-29细胞中HOTAIR m RNA的表达水平降低(P<0.01)。空白组与si-con组HT-29细胞侵袭数目、细胞划痕愈合率比较,差异无统计学意义(P>0.05);与si-con组比较,si-HOTAIR组HT-29细胞侵袭数目减少,细胞划痕愈合率降低(P<0.01)。与HOTAIR 3’UTR wt/miR-NC组比较,HOTAIR 3’UTR wt/miR-206组中HOTAIR wt的萤光素酶活性下降(P<0.01);而HOTAIR 3’UTR mut/miR-NC组与HOTAIR 3’UTR mut/miR-206组萤光素酶活性比较,差异无统计学意义(P>0.05)。与si-HOTAIR+anti-NC组比较,si-HOTAIR+anti-miR-206组中miR-206的表达水平降低(P<0.01)。与si-HOTAIR+anti-NC组比较,si-HOTAIR+anti-miR-206组中HT-29细胞的侵袭和迁移率增加(P<0.01)。结论HOTAIR通过负调控miR-206促进HT-29细胞的侵袭和迁移能力。Objective To investigate the effect of long noncoding RNA(lncRNA)HOX transcript antisense inter genic RNA(HOTAIR)targeting miR-206 on invasion and migration of colorectal cancer cells.Methods HT-29 cells of human colorectal cancer were cultured regularly,and the synthetic HOTAIR siRNA and siRNA control were transfected into HT-29 cells as si-HOTAIR group and si-con group,and the non-transfected HT-29 cells were set as blank group.Real time-quantitative polymerase chain reaction was used to determine the transfection effect,and Transwell and scratch healing tests were used to detect the invasion and migration of HT-29 cells.Bioinformatics software analyzed the binding sites of HOTAIR and miR-206 region,and constructed mutant(HOTAIR 3’UTR mut)and wild type(HOTAIR 3’UTR wt)luciferase reporter vectors.HOTAIR 3’UTR wt and HOTAIR 3’UTR mut were transfected into HT-29 cells with miR-206 mimic or miR-NC vector.HOTAIR 3’UTR wt/miR-NC group,HOTAIR 3’UTR wt/miR-206 group,HOTAIR 3’UTR mut/miR-NC group and HOTAIR 3’UTR mut/miR-206 group were cultured for 48 h to detect luciferase activity.HOTAIR siRNA and miR-206 inhibitor were co-transfected into HT-29 cells,si-HOTAIR+anti-NC group and si-HOTAIR+anti-miR-206 group were set up,and the cell invasion and migration levels were analyzed by Transwell and scratch healing assay.Results There was no significant difference in the expression level of HOTAIR mRNA in HT-29 cells between blank group and si-con group(P>0.05).Compared with si-con group,the expression level of HOTAIR mRNA in HT-29 cells in si-HOTAIR group was decreased(P<0.01).There was no significant difference in HT-29 cell invasion number and cell scratch healing rate between blank group and si-con group(P>0.05).Compared with the si-con group,the invasion number of HT-29 cells in the si-HOTAIR group was decreased,and the cell scratch healing rate was decreased(P<0.01).Compared with HOTAIR 3’UTR wt/miR-NC group,the luciferase activity of HOTAIR wt in the HOTAIR 3’UTR wt/miR-206 group decreased(P<0.01).T

关 键 词：结直肠癌 长链非编码RNA HOX转录反义RNA miR-206 细胞侵袭 细胞迁移 

分 类 号：R735.3[医药卫生—肿瘤]