长链非编码RNA ZEB1-AS1促进食管鳞癌侵袭转移

Long non-coding RNA ZEB1-AS1 promotes invasion and metastasis of esophageal squamous cell carcinoma

背景与目的:长链非编码RNA(long non-coding RNA,lncRNA)锌指E-盒结合同源异形盒1-反义链1(Zinc finger E-box binding homeobox 1 antisense 1,ZEB1-AS1)在多种肿瘤中高表达,与肿瘤患者临床病理学特征及预后相关,但其在食管鳞癌(esophageal squamous cell carcinoma,ESCC)中的作用及其机制尚不清楚。从细胞与分子生物学水平探讨lncRNA ZEB1-AS1在ESCC细胞侵袭转移中的作用。方法:通过实时荧光定量聚合酶链反应(realtime fluorescence quantitative polymerase chain reaction,RTFQ-PCR)方法检测9株ESCC细胞株中lncRNA ZEB1-AS1的表达水平,筛选出一株高表达细胞株。采用小干扰RNA(small interference RNA,siRNA)转染Eca-109细胞,分成干扰组(siZEB1-AS1)、干扰对照组(siNC)和空白组(Eca-109)。采用RTFQ-PCR方法检测lncRNA ZEB1-AS1的表达水平,采用细胞计数试剂盒(cell counting kit-8,CCK-8)实验检测细胞增殖能力情况,采用划痕实验和Transwell实验检测细胞迁移和侵袭能力情况,采用RTFQ-PCR方法和蛋白质印迹法(Western blot)检测ZEB1的表达水平。结果:9株ESCC细胞株中,Eca-109细胞中lncRNA ZEB1-AS1表达水平最高。siRNA抑制lncRNA ZEB1-AS1表达,降至对照组的57%。与对照组细胞相比,lncRNA ZEB1-AS1不影响Eca-109细胞增殖,但是能显著促进Eca-109细胞迁移和侵袭。lncRNA ZEB1-AS1上调ZEB1 mRNA和蛋白的表达。结论:lncRNA ZEB1-AS1通过上调ZEB1促进ESCC迁移、侵袭,lncRNA ZEB1-AS1/ZEB1或许可以作为ESCC治疗的潜在靶点。

Background and purpose:Long non-coding RNA Zinc finger E-box binding homeobox 1 antisense 1(lncRNA ZEB1-AS1)is highly expressed in various tumors,and is associated with clinicopathological features and prognosis of tumor patients.However,its role and mechanism in esophageal squamous cell carcinoma(ESCC)are still unclear.This study was to explore the role of lncRNA ZEB1-AS1 in invasion and metastasis of ESCC cells.Methods:The expression level of lncRNA ZEB1-AS1 in 9 ESCC cell lines was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR).Eca-109 cells were transfected with small interfering RNA(siRNA)and divided into interference group(siZEB1-AS1),interference control group(SiNC)and blank group(Eca-109).The expression level of lncRNA ZEB1-AS1 was detected by RTFQ-PCR.The cell proliferation ability was detected by cell counting kit-8(CCK-8)assay.The cell migration and invasion ability were detected by scratch assay and transwell experiment.The expression levels of ZEB1 mRNA and protein were detected by RTFQ-PCR and Western blot.Results:Among the 9 ESCC cell lines,the expression level of lncRNA ZEB1-AS1 was highest in Eca-109 cells.SiRNA inhibited lncRNA ZEB1-AS1 expression which decreased to 57%of that in the control group.Compared with the control cells,lncRNA ZEB1-AS1 did not affect Eca-109 cell proliferation,but significantly promoted Eca-109 cell migration and invasion.LncRNA ZEB1-AS1 up-regulated the expression levels of ZEB1 mRNA and protein.Conclusion:LncRNA ZEB1-AS1 promotes ESCC migration and invasion by upregulating ZEB1,and lncRNA ZEB1-AS1/ZEB1 may be a potential target for ESCC therapy.