Wang Peng-Li, Liu Bin, Xia Yang, Pan Chun-Feng, Ma Teng, Chen Yi-Jiang
Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Mol Med Rep. 2016 Apr;13(4):3074-82. doi: 10.3892/mmr.2016.4913. Epub 2016 Feb 19.
Long non-coding RNAs (lncRNAs) are involved in governing fundamental biological processes, and, in many lncRNAs, the expression level is altered and likely to have a functional role in tumorigenesis, including apoptosis, migration and invasion. The lncRNA‑Low Expression in Tumor (LET), a recently identified lncRNA, was demonstrated to be downregulated in hepatocellular and gallbladder cancer. However, its role in esophageal squamous cell carcinoma (ESCC) requires investigation. The expression level of lncRNA‑LET mRNA in primary ESCC and matched healthy tissues (48 cases) was determined by reverse transcription‑quantitative polymerase chain reaction. In addition, the effects of lncRNA-LET on cell apoptosis were evaluated by flow cytometric analysis, the regulatory effect of lncRNA‑LET on migration was detected using a wound healing assay and cellular invasion was analyzed by Matrigel‑coated transwell assay. Furthermore, the effect of lncRNA‑LET on cell proliferation was investigated by 5‑ethynyl‑2'-deoxyuridine cell proliferation assay and protein levels of lncRNA-LET targets were analyzed by western blotting. lncRNA-LET expression was decreased in primary ESCC tissues when compared with paired healthy tissues, and was identified to be associated with the clinical features. Overexpression of lncRNA‑LET was observed to inhibit the migration and invasion of ESCC cells, and modulate p53 expression levels in human ESCC cell lines in vitro. These results establish that lncRNA-LET is significant in the regulation of tumor progression and metastasis, and serves as a tumor suppressor in, and therefore has therapeutic potential for, the treatment of human ESCC.
长链非编码RNA(lncRNAs)参与调控基本的生物学过程,并且在许多lncRNAs中,其表达水平发生改变,可能在肿瘤发生过程中发挥功能性作用,包括细胞凋亡、迁移和侵袭。肿瘤低表达lncRNA(LET)是最近鉴定出的一种lncRNA,已证实在肝细胞癌和胆囊癌中表达下调。然而,其在食管鳞状细胞癌(ESCC)中的作用尚需研究。采用逆转录定量聚合酶链反应测定48例原发性ESCC及其配对的癌旁正常组织中lncRNA-LET mRNA的表达水平。此外,通过流式细胞术分析评估lncRNA-LET对细胞凋亡的影响,采用划痕愈合试验检测lncRNA-LET对迁移的调控作用,通过基质胶包被的Transwell试验分析细胞侵袭能力。此外,采用5-乙炔基-2'-脱氧尿苷细胞增殖试验研究lncRNA-LET对细胞增殖的影响,并通过蛋白质印迹法分析lncRNA-LET靶点的蛋白水平。与配对的正常组织相比,lncRNA-LET在原发性ESCC组织中的表达降低,且与临床特征相关。体外实验中,lncRNA-LET的过表达抑制了ESCC细胞的迁移和侵袭,并调节了人ESCC细胞系中p53的表达水平。这些结果表明,lncRNA-LET在肿瘤进展和转移的调控中具有重要作用,作为一种肿瘤抑制因子,对人类ESCC的治疗具有潜在的应用价值。