长链非编码RNA UBE2Q1-AS1对食管癌细胞增殖和迁移的调节作用及机制实验研究  

Regulatory effect and mechanism of lncRNA UBE2Q1-AS1 on proliferation and migration of esophageal cancer cells

作　　者：王银燕 黄翠 朱艳[1] 朱磊[2] WANG Yinyan;HUANG Cui;ZHU Yan;ZHU Lei(Department of Comprehensive Internal Medicine,Huangshi Maternal and Child Health Hospital,Huangshi 435000,China)

机构地区：[1]黄石市妇幼保健院综合内科,湖北黄石435000 [2]四川大学华西医院肿瘤科,四川成都610044

出　　处：《陕西医学杂志》2024年第9期1172-1176,1181,共6页Shaanxi Medical Journal

基　　金：国家自然科学基金资助项目(81902819)。

摘　　要：目的:观察长链非编码RNA(lncRNA)UBE2Q1-AS1对食管癌细胞增殖和迁移的影响,并探讨UBE2Q1-AS1发挥调节作用的分子机制。方法:实时荧光定量PCR(RT-qPCR)检测食管癌EC9706、Eca109、KYSE-510、TE-13细胞和永生化食管上皮HET-1A细胞中UBE2Q1-AS1的表达。将KYSE-510细胞分为si-NC组和si-UBE2Q1-AS1组,分别转染si-NC质粒和si-UBE2Q1-AS1质粒。集落形成实验检测各组KYSE-510细胞增殖活性。细胞划痕实验检测各组KYSE-510细胞的迁移能力。双荧光素酶报告系统验证UBE2Q1-AS1与微小RNA(miR)-377-3p的靶向关系。RT-qPCR检测各组KYSE-510细胞中miR-377-3p相对表达量。蛋白质印迹法(Western blot)检测各组KYSE-510细胞中同源盒蛋白A1(HOXA1)、纤连蛋白(Fibronectin)、叉头相关转录因子C2(FOXC2)、粒状蛋白质2同源物(GRHL2)和E-钙黏蛋白(E-cadherin)蛋白的相对表达量。结果:与HET-1A细胞比较,UBE2Q1-AS1在食管癌细胞中表达均升高(P<0.01)。si-UBE2Q1-AS1组KYSE-510细胞增殖活性低于si-NC组(P<0.01)。si-UBE2Q1-AS1组KYSE-510细胞迁移能力低于si-NC组(P<0.01)。UBE2Q1-AS1与miR-377-3p间存在靶向关系(P<0.01)。si-UBE2Q1-AS1组KYSE-510细胞中miR-377-3p表达水平高于si-NC组(P<0.01)。与si-NC组比较,si-UBE2Q1-AS1组KYSE-510细胞中HOXA1、Fibronectin、FOXC2蛋白表达降低,GRHL2和E-cadherin蛋白表达升高(均P<0.01)。结论:在食管癌细胞中,UBE2Q1-AS1表达上调,敲低UBE2Q1-AS1可能通过调节miR-377-3p/HOXA1轴抑制食管癌细胞增殖和迁移。Objective:To observe the effect of long non-coding RNA(lncRNA)UBE2Q1-AS1 on the proliferation and migration of esophageal cancer cells,and to explore the molecular mechanism by which UBE2Q1-AS1 exerts its regulatory effect.Methods:RT-qPCR was used to detect the expression of UBE2Q1-AS1 in esophageal cancer EC9706,Eca109,KYSE-510,TE-13 cells and immortalized esophageal epithelial HET-1A cells.KYSE-510 cells were divided into si-NC group and si-UBE2Q1-AS1 group,and were transfected with si-NC plasmid and si-UBE2Q1-AS1 plasmid respectively.Colony formation experiment was used to detect the proliferation activity of KYSE-510 cells in each group.The scratch experiment was used to detect the migration ability of KYSE-510 cells in each group.The dual-luciferase reporter system was used to verify the targeting relationship between UBE2Q1-AS1 and miR-377-3p.RT-qPCR was used to detecte the expression of miR-377-3p in KYSE-510 cells in each group.Western blot was used to detecte the expression of homeobox protein A1(HOXA1),fibronectin,forkhead box protein C2(FOXC2),grainyhead-like protein 2 homolog(GRHL2)and E-cadherin proteins in KYSE-510 cells in each group.Results:Compared with HET-1A cells,UBE2Q1-AS1 expression was increased in esophageal cancer cells(P<0.01).The proliferation activity of KYSE-510 cells in the si-UBE2Q1-AS1 group was lower than that in the si-NC group(P<0.01).The migration ability of KYSE-510 cells in the si-UBE2Q1-AS1 group was lower than that in the si-NC group(P<0.01).There was a targeting relationship between UBE2Q1-AS1 and miR-377-3p(P<0.01).The expression level of miR-377-3p in KYSE-510 cells in the si-UBE2Q1-AS1 group was higher than that in the si-NC group(P<0.01).Compared with the si-NC group,the protein expressions of HOXA1,Fibronectin,and FOXC2 in KYSE-510 cells in the si-UBE2Q1-AS1 group decreased,and the protein expressions of GRHL2 and E-cadherin increased(all P<0.01).Conclusion:UBE2Q1-AS1 expression is up-regulated in esophageal cancer cells,and UBE2Q1-AS1 down-regulation may inhibit the

关 键 词：食管癌 长链非编码RNA UBE2Q1-AS1 微小RNA-377-3p 细胞增殖 细胞迁移 

分 类 号：R73-3[医药卫生—肿瘤]