miR-1通过靶向LASP1和TAGLN2在食管鳞状细胞癌中的肿瘤抑制功能。

The tumor-suppressive function of miR-1 by targeting LASP1 and TAGLN2 in esophageal squamous cell carcinoma.

作者信息

Du Yan-Yan, Zhao Lian-Mei, Chen Liang, Sang Mei-Xiang, Li Jie, Ma Ming, Liu Jun-Feng

机构信息

Department of Clinical Laboratory, Fourth Hospital, Hebei Medical University, Shijiazhuang, China.

Scientific Research Center, Fourth Hospital, Hebei Medical University, Shijiazhuang, China.

出版信息

J Gastroenterol Hepatol. 2016 Feb;31(2):384-93. doi: 10.1111/jgh.13180.

DOI:10.1111/jgh.13180

PMID:26414725

Abstract

OBJECTIVE

This study determined the expression of microRNA-1 in esophageal squamous cell carcinoma (ESCC) tissue and cell lines to evaluate its effects on clinicopathological parameters and its target genes LASP1 and TAGLN2.

METHODS

The expression of miR-1, lasp1, and tagln2 was detected in 55 ESCC tissues and adjacent normal tissues by reverse transcription-polymerase chain reaction (RT-PCR). The association between miR-1, lasp1, and tagln2 expression and clinicopathological characteristics was observed. MicroRNA-1 (mimics-miR-1) and its inhibitor (Inhibitor-miR-1) were transfected into esophageal cancer cells KYSE 510 and Eca 109; cell proliferation, migration, and invasion assays were carried out. Plasmid construction and dual-luciferase reporter assay were also carried out to indicate whether LASP1 and TAGLN2 were miR-1 target genes. The expression of LASP1 and TAGLN2 was detected with Western blot methods in cell lines, by immunohistochemistry in ESCC tissue.

RESULTS

The gene expression level of microRNA-1 in cancer tissues was significantly lower than that in adjacent normal tissues (P < 0.01). The expression of miR-1 in ESCC was correlated with involvement of lymph nodes (P = 0.002), histologic classification (P = 0.000), and vessel invasion (P = 0.022). The expression of lasp1 and tagln2 increased in cancer tissues compared with in adjacent normal tissues (P < 0.05). MiR-1 suppresses the cell growth, migration, and invasion in vitro. The expression of LASP1 and TAGLN2 decreased in mimics-miR-1 transfected cells, and increased in inhibitor-miR-1 transfected cells. Luciferase reporter assay confirmed that LASP1 and TAGLN2 mRNA actually had the target sites of miR-1.

CONCLUSIONS

miR-1 suppresses cell proliferation, invasiveness, metastasis, and progression of ESCC by binding its targeted genes LASP1 and TAGLN2.

摘要

目的

本研究旨在测定微小RNA - 1在食管鳞状细胞癌（ESCC）组织及细胞系中的表达，以评估其对临床病理参数的影响及其靶基因LASP1和TAGLN2。

方法

采用逆转录 - 聚合酶链反应（RT - PCR）检测55例ESCC组织及其癌旁正常组织中miR - 1、lasp1和tagln2的表达。观察miR - 1、lasp1和tagln2表达与临床病理特征之间的关联。将微小RNA - 1（模拟物 - miR - 1）及其抑制剂（抑制剂 - miR - 1）转染至食管癌细胞KYSE 510和Eca 109；进行细胞增殖、迁移和侵袭实验。还进行了质粒构建和双荧光素酶报告基因检测，以确定LASP1和TAGLN2是否为miR - 1的靶基因。采用蛋白质免疫印迹法检测细胞系中LASP1和TAGLN2的表达，采用免疫组织化学法检测ESCC组织中的表达。

结果

癌组织中微小RNA - 1的基因表达水平显著低于癌旁正常组织（P < 0.01）。ESCC中miR - 1的表达与淋巴结转移（P = 0.002）、组织学分级（P = 0.000）和血管侵犯（P = 0.022）相关。与癌旁正常组织相比，癌组织中lasp1和tagln2的表达增加（P < 0.05）。MiR - 1在体外抑制细胞生长、迁移和侵袭。模拟物 - miR - 1转染的细胞中LASP1和TAGLN2的表达降低，抑制剂 - miR - 1转染的细胞中表达增加。荧光素酶报告基因检测证实LASP1和TAGLN2 mRNA确实具有miR - 1的靶位点。