目的：探讨长链非编码RNA（lncRNA）LINC01140在食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）组 织及细胞中的表达及其对Eca109细胞增殖与侵袭的影响及其分子机制。方法：选取2012年3月至2015年5月河北医科大学第 四医院收治的 133 例 ESCC 患者的临床资料和 GEPIA 数据库中收集的 182 例 ESCC 组织及 286 例食管正常黏膜组织的 LINC01140表达数据，以及 ESCC 细胞系 Kyse150、Eca109 和 TE13。用 qPCR 法检测癌组织和细胞中LINC01140的表达水平， 分析其表达水平与患者临床病理特征及预后的关系。分别将pcDNA3.1-LINC01140、阴性对照（pcDNA3.1-NC）或 miR-452-5p mimic及阴性对照（miR-NC）转染到Eca109细胞，MTS、Transwell实验分别检测细胞的增殖与侵袭能力。用双荧光报告基因实验 及 TOP/FOP 报告基因系统检测 LINC01140 与 miR-452-5p 的靶向结合作用及 LINC01140 对 Wnt/β-catenin 通路活化水平的影 响。结果：LINC01140在ESCC组织和细胞中表达均显著下调（均P<0.01），LINC01140 低表达与 ESCC 患者年龄、淋巴结 转移、TNM分期及OS密切相关（均P<0.05）。LINC01140过表达明显抑制 Eca109 细胞的增殖及侵袭能力（均P<0.01）。机制 研究表明，LINC01140可能通过竞争结合miR-452-5p影响Wnt/β-catenin信号通路的活化水平继而调控Eca109细胞的恶性生物学 行为。结论：LINC01140通过靶向miR-452-5p/Wnt/β-catenin轴促进ESCC细胞的增殖与侵袭能力，其有望成为ESCC患者靶向 治疗的潜在靶点及预后评估的标志物。
Objective: To investigate the expression of lncRNA LINC01140 in esophageal squamous cell carcinoma (ESCC) tissues and cell lines, and to explore its effect on the proliferation and invasion of Eca109 cells as well as the possible molecular mechanism. Methods: The clinical data of 133 ESCC patients who were treated in the Fourth Hospital of Hebei Medical University from March 2012 to May 2015 and the data of 182 ESCC tissues and 286 normal esophageal mucosa tissues included in GEPIA database, as well as ESCC cell lines (Kyse150, Eca109, TE13) were collected for this study. The expression level of LINC01140 in ESCC tissues and cell lines was detected by qPCR method, and the relationship between its expression level and clinicopathological features as well as the prognosis of ESCC patients was further analyzed. pcDNA3.1-LINC01140, negative control (pcDNA3.1-NC) or miR-452-5p mimics and negative control (miR-NC) were respectively transfected into Eca109 cells. Then, MTS and Transwell assay were performed to assess the effect of LINC01140 on proliferation and invasion of transfected cells. The interaction between LINC01140 and miR-452-5p, as well as the effect of LINC01140 on the activation of Wnt/β-catenin pathway was detected by Dual-luciferase reporter gene assay and TOP/FOP reporter gene system. Results: The expression of LINC01140 was downregulated in ESCC tissues and cell lines (all P<0.01). Low expression of LINC01140 was closely correlated with the age, lymph node metastasis, TNM stage and the OS of ESCC patients (all P <0.01). Overexpression of LINC01140 significantly reduced the proliferation and invasion ability of Eca109 cells (all P<0.01). Mechanistic analysis indicated that LINC01140 affected the activation of Wnt/β-catenin signaling pathway possibly via competitively sponging miR-452-5p to further regulate the malignant biological behaviors of Eca109 cells. Conclusion: LINC01140 affects proliferation and invasion of ESCC cells via regulating miR-452-5p/Wnt/β-catenin axis. LINC01140 is expected to be a potential target for the targeted therapy and the molecular marker for the prognosis evaluation of ESCC patients.
河北省自然科学基金资助项目（No. H2020206368）；河北省医学科学研究重点课题计划项目（No. 20210399）；河北省人才工程培养 资助项目（No. 201901035）