Gan Lu, Chen Yan, Liu Hua, Ju Wen-Hui
Department of Obstetrics and Gynecology, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Hubei Province, Jingzhou, China.
Department of Obstetrics and Gynecology, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Hubei Province, Jingzhou, China,
Gynecol Obstet Invest. 2019;84(2):136-144. doi: 10.1159/000493265. Epub 2018 Sep 25.
To investigate whether long non-coding RNA (lncRNA) ZEB1 antisense 1 (ZEB1-AS1) affects cell migration and invasion of cervical cancer by regulating epithelial-mesenchymal transition (EMT) via the p38MAPK pathway.
Human cervical cancer cell line Hela was classified into Control, NC siRNA, ZEB1-AS1 siRNA, SB203580 (p38MAPK pathway inhibitor) and ZEB1-AS1 siRNA + Anisomycin (p38MAPK pathway activator) groups. Quantitative real-time polymerase chain reaction was performed for ZEB1-AS1 expression, Western blotting to measure p38MAPK signaling pathway-/EMT-related proteins, and Wound-healing and Transwell assays to evaluate cell migration and invasion respectively.
ZEB1-AS1 was upregulated in cancer tissues and related to major clinicopathological features of cervical cancer. Besides, patients with lower-ZEB1-AS1-expression had a higher 5-year survival rate than those patients with higher-ZEB1-AS1-expression. High ZEB1-AS1 expression and advanced Federation of Gynecology and Obstetrics stage were independent risk factors for patients' prognosis. Both ZEB1-AS1 siRNA and SB203580 effectively reduced p-p38 expression and the migration and invasion of Hela cells, with elevation of E-cadherin and reduction of Vimentin and N-cadherin. However, inhibitory effects of ZEB1-AS1 siRNA on EMT as well as cell migration and invasion of the Hela cell were reversed by Anisomycin.
Inhibition of ZEB1-AS1 can block the p38MAPK signaling pathway, ultimately restricting the EMT and suppressing cell migration and invasion of cervical cancer cells.
探讨长链非编码RNA（lncRNA）ZEB1反义RNA1（ZEB1-AS1）是否通过p38丝裂原活化蛋白激酶（p38MAPK）信号通路调控上皮-间质转化（EMT），进而影响宫颈癌细胞的迁移和侵袭。
将人宫颈癌Hela细胞系分为对照组、阴性对照小干扰RNA（NC siRNA）组、ZEB1-AS1 siRNA组、SB203580（p38MAPK信号通路抑制剂）组和ZEB1-AS1 siRNA+茴香霉素（p38MAPK信号通路激活剂）组。采用定量实时聚合酶链反应检测ZEB1-AS1表达，蛋白质印迹法检测p38MAPK信号通路及EMT相关蛋白，划痕实验和Transwell实验分别评估细胞迁移和侵袭能力。
ZEB1-AS1在癌组织中表达上调，且与宫颈癌主要临床病理特征相关。此外，ZEB1-AS1低表达患者的5年生存率高于高表达患者。ZEB1-AS1高表达和国际妇产科联盟分期晚期是患者预后的独立危险因素。ZEB1-AS1 siRNA和SB203580均能有效降低p-p38表达以及Hela细胞的迁移和侵袭能力，同时使E-钙黏蛋白表达升高，波形蛋白和N-钙黏蛋白表达降低。然而，茴香霉素可逆转ZEB1-AS1 siRNA对EMT以及Hela细胞迁移和侵袭的抑制作用。
抑制ZEB1-AS1可阻断p38MAPK信号通路，最终抑制EMT并抑制宫颈癌细胞的迁移和侵袭。
