长链非编码 RNA H19 通过 STAT3/EZH2 轴促进食管癌的上皮间质转化和转移。
LncRNA H19 promotes epithelial mesenchymal transition and metastasis of esophageal cancer via STAT3/EZH2 axis.
机构信息
Department of Thoracic Surgery, The Second Xiangya Hospital of Central South University, Changsha 410011, PR China.
Department of Respiratory Medicine, The First Hospital of Changsha, Changsha 410005, PR China.
出版信息
Int J Biochem Cell Biol. 2019 Aug;113:27-36. doi: 10.1016/j.biocel.2019.05.011. Epub 2019 May 16.
BACKGROUND
Long non-coding RNA H19 (lncRNA H19) has been widely reported in esophageal cancer (EC), and previous study had found that lncRNAH19 was up-regulated in EC and promoted cell proliferation and metastasis. However, the mechanism still needs further studied.
METHODS
Levels of lncRNA H19 were analyzed by qRT-PCR in matched samples from 30 patients. Expression levels of lncRNA H19, let-7, STAT3 and EZH2 were additionally identified by qRT-PCR and western blotting in five EC cell lines. The effects of lncRNA H19 on cell proliferation, migration, invasion and apoptosis in cell lines were performed by MTT assay, colony formation assay, Transwell assay and flow cytometry in vitro, and tumor formation was detected by xenograft nude mice model in vivo. The expression level of STAT3, EZH2, β-catenin, and EMT and metastasis related molecules such as E-cadherin, N-cadherin, Snail-1 and MMP-9 was assessed by qRT-PCR and western blotting. Finally, luciferase reporter assay and RIP assay were used to verify the interaction between lncRNA H19 and let-7c, and their subsequent regulation of STAT3.
RESULTS
Knockdown of lncRNA H19 repressed cell proliferation, migration and invasion as well as EMT and metastasis via STAT3-EZH2-β-catenin pathway, while lncRNA H19 regulated STAT3 negatively regulated let-7c in EC cell lines.
CONCLUSIONS
lncRNA H19 facilitates EMT and metastasis of EC through let-7c/STAT3/EZH2/β-catenin axis.
背景
长链非编码 RNA H19（lncRNA H19）在食管癌（EC）中广泛报道，先前的研究发现 lncRNAH19 在 EC 中上调，并促进细胞增殖和转移。然而，其机制仍需要进一步研究。
方法
在 30 名患者的配对样本中通过 qRT-PCR 分析 lncRNA H19 的水平。通过 qRT-PCR 和 Western blot 分别鉴定 5 种 EC 细胞系中 lncRNA H19、let-7、STAT3 和 EZH2 的表达水平。体外 MTT 测定、集落形成测定、Transwell 测定和流式细胞术检测 lncRNA H19 对细胞系增殖、迁移、侵袭和凋亡的影响，体内裸鼠异种移植模型检测肿瘤形成。通过 qRT-PCR 和 Western blot 评估 STAT3、EZH2、β-catenin 以及 EMT 和转移相关分子如 E-钙粘蛋白、N-钙粘蛋白、Snail-1 和 MMP-9 的表达水平。最后，通过荧光素酶报告基因检测和 RIP 检测验证 lncRNA H19 与 let-7c 的相互作用及其对 STAT3 的后续调控。
结果
lncRNA H19 的敲低通过 STAT3-EZH2-β-catenin 通路抑制细胞增殖、迁移和侵袭以及 EMT 和转移，而 lncRNA H19 在 EC 细胞系中负调控 STAT3 下调 let-7c。
结论
lncRNA H19 通过 let-7c/STAT3/EZH2/β-catenin 轴促进 EC 的 EMT 和转移。
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